ABCMdb

ABCC2 p.Ile1173Phe

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Publications

PMID: 18464048 [PubMed] Gradhand U et al: "Pharmacogenomics of MRP transporters (ABCC1-5) and BCRP (ABCG2)."

No. Sentence Comment

101 Several molecular defects in MRP2 have been suggested to result in DJS including those which produce deficient protein maturation (Hashimoto et al., 2002; Keitel et al., 2003), proteasomal degradation (Keitel, 2003), impaired membrane sorting (Hashimoto et al., 2002; Mor-Cohen et al., 2001), loss in transport activity (Mor-Cohen et al., 2001), Figure 2 Predicted membrance topology of MRP2 (ABCC2) based on hydrophobicity analysis. Locations of the non-synonymous polymorphisms are indicated with arrows. See Table 2 for allele frequencies and description of funtional consequences. NH2 COOH NBD NBD in out Membrane Pro19Leu Phe39Tyr Arg100* Arg100Gln Ser281Asn Ser325* Asp333Gly Arg353His Arg412Gly Val417Ile Lys430Arg Thr486Ile Gly676Arg Trp709Arg Asn718Ser Ser789Phe Arg768Trp Asp833Asn Glu893Gln Leu927Arg Lys961Arg Tyr967* Phe981Leu Gln1019His Arg1066* Arg1150His Arg1100Cys Arg1100His Ile1137Phe Ile1173Phe Val1188Glu Arg1174His Arg1181Leu Asn1244Lys Thr1273Ala Pro1291Leu Lys1299Gln Arg1310* Ser1367Cys Gln1382Arg Arg1392del Met1393del Ala1450Thr Thr1476Met Cys1515Tyr MRP2 (ABCC2) NBD NBD Asp833Asn Glu893Gln Leu927Arg Lys961Arg Tyr967* NBD NBDNBD Asp833Asn Glu893Gln Leu927Arg Lys961Arg Tyr967* 325 Table2MRP2(ABCC2)singlenucleotidepolymorphisms.Location,allelefrequencyandfunctionaleffects. Positionin codingsequence Amino acidexchangeLocation Allelefrequency EffectNCBIIDReferenceAfCaJpothers 56C>TPro19LeuExon2--1[1]b -- 116T>APhe39TyrExon2--0[2]--rs927344 298C>TArg100*Exon3--[3]-DJS[3] 299G>AArg100GlnExon3--1[1]b -- 842G>ASer281AsnExon7-0[4]1[1]b -- 974C>GSer325*Exon8---Malayan[5]DJS[5] 998A>GAsp333GlyExon8--0[2]--rs17222674 1058G>AArg353HisExon9--0[2]--rs7080681 1271A>GArg412GlyExon10-[6]0[2]-DJS;Decreaseinmethotrexateelimination[6] 1249G>AVal417IleExon10-22[7]13[9]-lowermRNAand(protein)expressioninpreterm placenta[11] rs2273697 26[8]16[4]noeffectonRNAandproteinininduodenum[12] 19[10]noeffectonproteininliver[8] noeffectonconjugatedbilirubinlevelinserum[13] changesinlocalizationinneuroepithelialtumors[14] possibleassociationwithtenofovir-inducedrenal proximaltubulopathy[15] 1289A>GLys430ArgExon10-4[16]0[2]-- 1457C>TThr486IleExon10-0[4]3[1]b -- 2026G>CGly676Arg--0[2]-DJS[17] 2125T>CTrp709Arg--0[2]-DJS[17] 2153A>GAsn718SerExon17-0[4]0[2]--rs3740072 2302C>TArg768TrpExon18-0[18]1[9]-DJS;deficientmaturationandimpairedsorting[19] 2366C>TSer789PheExon18-0[18]1[9]-lowerexpressionandmembranelocalization[20] noeffectonconjugatedbilirubinlevelinserum[13]/ heterozygous 2647G>AAsp883AsnExon20--1[1]b -- 2677G>CGlu893GlnExon20--0[2]--rs3740071 2780T>GLeu927ArgExon21-1[10]0[2]-- (Continued) Table2(Continued) Positionin codingsequence Aminoacid exchangeLocation Allelefrequency EffectNCBIIDReferenceAfCaJpothers 2882A>GLys961ArgExon21--1[1]b --- 2901C>ATyr967*Exon22--0[2]--rs17222547 2943C>GPhe981LeuExon22-2[21]0[2]-Noinfluenceonpravastatinkinetics[21] 3057G>TGln1019HisExon22--1[1]b -- 3196C>TArg1066*Exon23-[22]0[2]-DJS;truncatedprotein[22][23] 3298C>TArg1100CysExon24-1[10]0[2]-- 3299G>AArg1100HisExon24-1[10]0[2]-- 3449G>AArg1150HisExon25--0[2]Israeli[24]DJS;impairedtransportactivityintransfectedcells althoughnormalexpressionandlocalization[24] 3517A>TIle1173PheExon25--0[2]Israeli[24]DJS;impairedproteinmaturationandproteasomal degradation[25] lowexpression,mislocation,andimpairedtransport activityintransfectedcells[24] 3521G>AArg1174HisExon25-0[4]1[1]b -- 3542G>TArg1181LeuExon25-0[4]0[2]--rs8187692 3563T>AVal1188GluExon25-7[4]1[1]b -noeffectonnelfinaviraccumulationinPBMC[4],rs17222723 4[16]associatedwithanthracycline-induced cardiotoxicity[26] 6[8] 3732C>TAsn1244LysExon26--0[1]b -- 0[2] 3817A>GThr1273AlaExon27--0[2]--rs8187699 3872C>TPro1291LeuExon28--0[2]--rs17216317 3897A>CLys1299GlnExon28--0[2]--rs4148400 3928C>TArg1310*Exon28--0[2]-DJS[17,27] 4100C>GSer1367CysExon29--1[1]b -- 4145A>GGln1382ArgExon29--[28]-DJS;noeffectonmaturationorsorting,impaired substrate-inducedATPhydrolysis[19] 4175-80delArg1392delExon30--0[2]-DJS;deficientMRP2maturationandimpaired sortingtoapicalmembraneintransfectedcells[29] 327 4348G>AAla11450ThrExon31-0[18]1[9]-lowerexperssionandmembracelocalizationin transfectedcells[20] 4461C>TThr1476MetExon31-[30]1[2]-- 4544G>ACys1515TyrExon32-9[4]1[1]b -noeffectonnelfinaviraccumulationinPBMC[4]rs8187710 5[10]associatedwithanthracycline-induced cardiotoxicity[26] 4[16] 6[8] ReferencewithoutfrequencymeansthatSNPwasdetectedbutnofrequencydetermined. Login to comment

PMID: 18673259 [PubMed] Nakamura T et al: "Pharmacogenetics of intestinal absorption."

No. Sentence Comment

33 Two mutations 3517A>T and 3449G>A, predicting the substitution Ile1173Phe and Arg1150His, respectively, impaired the transport of carboxyfluorescein by ABCC2, while the 3517A>T mutation induced weaker ABCC2 expression and mislocation to the ER [39, 41]. Login to comment

PMID: 14965249 [PubMed] Haimeur A et al: "The MRP-related and BCRP/ABCG2 multidrug resistance proteins: biology, substrate specificity and regulation."

No. Sentence Comment

380 Two other DJS mutations that are not in the NBDs are Arg1150His and Ile1173Phe [276, 277]. Login to comment
382 In contrast, the MRP2-Ile1173Phe mutant protein was both mislocalized and non-functional. Login to comment
383 The Ile1173Phe and Arg1150His mutations are both in MSD3 but their precise location varies according to different topological models of the protein. Login to comment

PMID: 16006996 [PubMed] Conseil G et al: "Polymorphisms of MRP1 (ABCC1) and related ATP-dependent drug transporters."

No. Sentence Comment

167 The G3449A mutation (which results in a substitution of His for Arg1150 ) is found in Moroccan- Jewish DJS patients whereas the MRP2-Ile1173Phe (A3517T) mutation is mainly found in Iranian-Jewish patients. Login to comment
168 Although both the Arg1150His and Ile1173Phe mutations eliminate MRP2 transport activity in vitro, the Ile1173Phe variant protein also exists predominantly in an underglycosylated form that is retained in the endoplasmic reticulum [47,48]. Login to comment

PMID: 16041239 [PubMed] Colombo S et al: "Influence of ABCB1, ABCC1, ABCC2, and ABCG2 haplotypes on the cellular exposure of nelfinavir in vivo."

No. Sentence Comment

71 - 24C > T exon 1 Itoda et al., 2002 mp-v-004 rs717620 IVS 6-30 G > T intron 6 Epidauros mp-v-051 rs8187666 c.842G > A exon 7 p.S281N Epidauros mp-v-115 c.998G > A intron 7 Epidauros mp-v-083 c.1219C > T exon 10 synonymous (p.L407L) Epidauros mp-v-007 rs8187669 c.1249G > A exon 10 p.V417I Itoda et al., 2002 mp-v-008 rs2273697 c.1346C > G exon 10 synonymous (p.T482T) Epidauros mp-v-114 c.1457C > T exon 10 p.T486I Epidauros mp-v-055 rs8187670 IVS 16 - 47 G > A intron 16 Epidauros mp-v-118 IVS 16 - 30 T > A intron 16 Epidauros mp-v-119 c.2153A > G exon 17 p.N718S Epidauros mp-v-093 rs3740072 c.2216T > C exon 17 p.L739P Epidauros mp-v-108 c.3449G > A exon 25 p.R1150H Mor-Cohen et al., 2001 mp-v-085 c.3517A > T exon 25 p.I1173F Keitel et al., 2003 mp-v-096 c.3521G > A exon 25 p.R1174H Epidauros mp-v-068 c.3542G > T exon 25 p.R1181L Epidauros mp-v-069 rs8187692 c.3563T > A exon 25 p.V1188E Epidauros mp-v-025 rs8187694 IVS 30 - 53 C > T intron 30 Epidauros mp-v-105 rs3824610 c.4348G > A exon 31 p.A1450T Suzuki et al. 2002 mp-v-106 c.4410G > A exon 31 synonymous (p.E1470E) Epidauros mp-v-077 rs8187706 c.4488C > T exon 31 synonymous (p.H1496H) Epidauros mp-v-038 rs8187707 IVS 31 + 12 G > A intron 31 Epidauros mp-v-039 rs8187708 IVS 31 + 74 C > T intron 31 Epidauros mp-v-040 IVS 31 - 9 T > C intron 31 Epidauros mp-v-042 c 4527C > T exon 32 synonymous (p.A1509A) Epidauros mp-v-048 rs8187709 c.4544G > A exon 32 p.C1515Y Epidauros mp-v-043 rs8187710 + 259 G > T 30 flanking Epidauros mp-v-120 Transporter polymorphisms and HIV treatment Colombo et al. 601 BCRP (ABCG2) g. Login to comment

PMID: 22330094 [PubMed] Sekine S et al: "Sustained intrahepatic glutathione depletion causes proteasomal degradation of multidrug resistance-associated protein 2 in rat liver."

No. Sentence Comment

19 Certain DJS mutations (e.g., missence mutations R768W and I1173F; deletion mutation R1392, M1393) have been reported to cause defects in canalicular sorting and rapid proteasomal degradation of the hMRP2 protein [2-4]. Login to comment

PMID: 22565165 [PubMed] Grover S et al: "Genetic association analysis of transporters identifies ABCC2 loci for seizure control in women with epilepsy on first-line antiepileptic drugs."

No. Sentence Comment

89 - 24C > T 50 UTR k Promoter activity [haplotype containing (- 1549A)-(- 24T)] [33] k mRNA expression [29] m Expression and activity [35] m Expression [haplotype containing (- 24C)-1249A- 3972C] [36] k Expression [haplotype containing (- 24C)- 1249G- 3972T, (-24T)-1249G- 3972C or (-24T)-1249G- 3972T] [36] k Clearance of mycophenolic acid [35] k Clearance of methotrexate [37] k Clearance of irinotecan (ABCC2*2 containing the wild-type C allele) [29] 0.137 5 rs4919395 chr10:101542963 c.33 + 329G > A Intron 1 0.384 6 rs2756104 chr10:101544026 c.34 - 339C > T Intron 1 0.392 7 rs927344 chr10:101544447 c.116T > A Exon 2 (Phe39Tyr) 0.000 8 rs4148385 chr10:101548177 c.207+ 3639C > A Intron 2 0.382 9 rs2180990 chr10:101548974 c.208 - 3017C > G Intron 2 0.392 10 rs35191126 chr10:101549533:34 c.208 - 2458_208 - 2457G > delG Intron 2 0.384 11 rs4148389 chr10:101549911 c.208 - 2080A > G Intron 2 0.396 12 rs2804400 chr10:101553259 c.334 - 49C > T Intron 3 0.394 13 rs2756109 chr10:101558746 c.868 - 218T > G Intron 7 0.364 14 rs7080681 chr10:101560169 c.1058G > A Exon 9 (Arg353His) 0.000 15 rs2273697 chr10:101563815 c.1249G > A Exon 10 (Val417Ile) m mRNA expression [38] m Expression [haplotype containing (- 24C)-1249A- 3972C] [36] k Expression [haplotype containing (- 24C)- 1249G- 3972T, (-24T)-1249G- 3972C or (-24T)-1249G- 3972T] [36] k Clearance of irinotecan (ABCC*2 containing G allele) [34] 0.271 16 rs113646094 chr10:101564012 c.1446C > G Exon 10 (Thr482Thr) m mRNA expression [39] 0.002 17 rs2073337 chr10:101567426 c.1668 + 148A > G Intron 12 0.388 18 rs2756114 chr10:101569483 c.1816 - 408T > C Intron 13 0.393 19 rs3740074 chr10:101571528 c.1967+ 169T > C Intron 15 0.368 20 rs4148394 chr10:101572343 c.1968 - 432A > C Intron 15 0.206 21 rs3740072 chr10:101577123 c.2153A > G Exon 17 (Asn718Ser) 0.000 22 rs56199535 chr10:101578577 c.2302C > T Exon 18 (Arg768Trp) 0.000 23 rs56220353 chr10:101578641 c.2366C > G/T Exon 18 (Ser789Cys/Phe) k Activity, k expression and impaired membrane localization [40] 0.000 24 rs2002042 chr10:101587931 c.2621 - 2133C > T Intron 19 0.204 25 rs11442349 chr10:101589215:16 c.2621 - 849_2621 - 848T > delT Intron 19 0.375 26 rs3740071 chr10:101590120 c.2677C > G Exon 20 (Gln893Glu) 0.000 27 rs7898096 chr10:101593385 c.3259 -752G > A Intron 23 0.000 28 rs17216345 chr10:101594274 c.3396T > C Exon 24 (Ile1132Ile) 0.027 29 rs72558200 chr10:101595882 c.3449 G > A Exon 25 (Arg1150His) 0.000 30 rs72558201 chr10:101595950 c.3517 A > T Exon 25 (Ile1173Phe) 0.000 31 rs8187692 chr10:101595975 c.3542G > T Exon 25 (Leu1181Arg) 0.000 32 rs17222723 chr10:101595996 c.3563T > A Exon 25 (Val1188Glu) m Expression [41] 0.016 ABCC2 polymorphism and response to AEDs Grover et al. 451 or liver functioning. Login to comment

PMID: 22290738 [PubMed] Arlanov R et al: "Functional characterization of protein variants of the human multidrug transporter ABCC2 by a novel targeted expression system in fibrosarcoma cells."

No. Sentence Comment

169 Computational comparative genomic studies using PolyPhen 2 predicted that nine of these ABCC2 variants (F39Y, D333G, I670T, I1036T, R1174H, R1181L, V1188E, N1244K, and P1291L) would be located within the transmembrane regions, close to the ATP-binding domain of ABCC2 or close to two missense variants (I1173F, R1150H) causing Dubin-Johnson syndrome [Keitel et al., 2003; Mor-Cohen et al., 2001]. Login to comment
174 The Dubin-Johnson syndrome- causing variant ABCC2 I1173F served as a positive control for reduced ABCC2 expression. Login to comment
177 Staining of ABCC2 was weaker for D333G, I1173F, R1174H, and Table 1. Login to comment
272 The selected variants R1174H, R1181L, and V1188E (linked to C1515Y) from the present study, in addition to two recently identified variants that cause Dubin-Johnson syndrome (R1150H, I1173F), are located at the same gene region between the transmembrane helices TM15 and TM16. Login to comment

PMID: 20082599 [PubMed] Jemnitz K et al: "ABCC2/Abcc2: a multispecific transporter with dominant excretory functions."

No. Sentence Comment

86 The I1173F mutation showed a similar defect in trafficking (Keitel et al., 2003). Login to comment
97 Mutant Predicted location Substrate Activity changes Reference Human MRP2 Δ1-188 TMD0 LTC4 ↓ Fernandez et al., 2002 K316A JC, TM6 GMF ↔ Ryu et al., 2000 K324A TM6 GMF ↓ Ryu et al., 2000 K329A TM6 GMF ↔ Ryu et al., 2000 R412G DJ IC MTX ↓ Hulot et al., 2005 W417I IC, TM7-TM8 E2-17βG ↓ Hirouchi et al., 2004 LTC4 ↓ DNP-SG ↓ H439A TM8 GMF ↔ Ryu et al., 2000 K483A IC, JM, TM9 GMF ↓ Ryu et al., 2000 K590A JC, TM11 GMF ↔ Ryu et al., 2000 S789F NBD1 E2-17βG ↓ Hirouchi et al., 2004 LTC4 ↓ DNP-SG ↓↓ R1023A EC, JM, TM13 GMF ↔ Ryu et al., 2000 H1042A TM13 GMF ↔ Ryu et al., 2000 R1100A JC, TM14 GMF ↔ Ryu et al., 2000 P1158A IC, JM, TM15 LTC4 ↓↓ Letourneau et al., 2007 E2-17βG ↔ MTX ↔ Table 1. continued on next page Mutant Predicted location Substrate Activity changes Reference I1173F DJ IC, TM15-16 LTC4 No act Keitel et al., 2003 E2-17βG No act R1210A EC, JC, TM16 GMF ↓↓ Ryu et al., 2000 R1230A TM16 GMF ↔ R1257A JC, TM17 GMF ↓↓ W1254A JC, TM17 E2-17βG ↓↓ Ito et al., 2001a W1254C ↓↓ W1254F ↔ W1254Y ↔ W1254A JC, TM17 LTC4 ↓↓ Ito et al., 2001b W1254C ↓↓↓ W1254F ↓↓ W1254Y ↓↓ W1254A JC, TM17 MTX ↓↓ Ito et al., 2001a W1254C ↓↓ W1254F ↓↓ W1254Y ↓↓↓ A1450T NBD2 E2-17βG ↓↓ Hirouchi et al., 2004 LTC4 ↓↓ DNP-SG ↓↓ Rat Mrp2 K308M IC, JM, TM6 TLC-S ↔ Ito et al., 2001b DNP-G ↑ LTC4 ↓ E3040G ↔ K320M TM6 TLC-S ↑ DNP-G ↑ LTC4 ↓ E3040G ↑ K325M TM6 TLC-S ↓* DNP-G ↓↓↓* LTC4 ↓↓↓* E3040G ↓ D329N TM6 TLC-S ↔ DNP-G ↓ LTC4 ↓↓↓* E3040G ↓ R586L TM11 TLC-S ↓ DNP-G ↓↓* LTC4 ↓↓* E3040G ↔ R1019M IC, JM, TM13 TLC-S ↔ DNP-G ↑* LTC4 ↔ E3040G ↔ R1096L TM14 TLC-S ↑ DNP-G ↑ LTC4 ↔ E3040G ↔ EC, extracellular; IC, intracellular; JC, near the cytosol in the membrane; JM, juxtamembrane; TLC-S, tauro-litocholate-sulfate; GMF, glutathione- methyl-fluorescein; ↑, activity over control>1.2; ↔, 1.2>activity over control>0.8; ↓, 0.8>activity over control>0.5; ↓↓, 0.5>activity over control>0.1; ↓↓↓, 0.1>activity over control. Login to comment
102 Similarly, deletion of amino acids 1-188 (Fernandez et al., 2002), the R412G change (Hulot et al., 2005), and an I1173F replacement (Keitel et al., 2003) in two Dubin-Johnson variants impaired leukotriene-C4 (LTC4) transport. Login to comment
113 Among these mutations recently compiled by Nies and Keppler (2007), of the five amino acids affected that are located outside the nucleotide-binding domains, four are basic (R100X, R393W, R1066X, and R1150H) and one is neutral (I1173F). Login to comment
115 I1173F is improperly processed and degraded, but has been shown in membrane assays as nonfunctional (Keitel et al., 2003). Login to comment

PMID: 17287630 [PubMed] Mor-Cohen R et al: "Age estimates of ancestral mutations causing factor VII deficiency and Dubin-Johnson syndrome in Iranian and Moroccan Jews are consistent with ancient Jewish migrations."

No. Sentence Comment

2 FVII deficiency in both populations is caused by a founder A244V mutation in the F7 gene and DJS is caused by two founder mutations, I1173F and R1150H in the MRP2 gene that are specific for Iranian and Moroccan Jewish patients, respectively. Login to comment
3 We estimated the age of FVII A244V and MRP2 I1173F by analysis of microsatellite markers flanking F7 and MRP2 genes, respectively, in 13 Iranian Jewish homozygotes for the I1173F mutation and 21 Iranian and Moroccan Jewish homozygotes for the A244V mutation. Login to comment
5 The estimated age of the I1173F mutation was approximately 1500 years, and the age of the A244V mutation was approximately 2600 years. These estimates suggest that I1173F causing DJS in Iranian Jews occurred after the separation of Iranian Jews from Moroccan Jews 2000-2600 years ago, while A244V causing FVII deficiency in Iranian and Moroccan Jews occurred prior to the divergence of these two populations. Login to comment
19 We previously reported two distinct missense mutations, I1173F and R1150H, causing DJS in Iranian and Moroccan Jewish patients, respectively. Login to comment
24 In this study we addressed the question of whether an age estimate of the FVII A244V mutation would be consistent with the hypothesis that this mutation occurred before the historic separation of Moroccan from Iranian Jews, and that the MRP2 I1173F mutation causing DJS in Iranian Jews occurred after the divergence of the Iranian and Moroccan Jews. Login to comment
37 Age estimates of the MRP2 I1173F and FVII A244V mutations Estimating the age of the MRP2 I1173F mutation and the FVII A244V mutation was carried out by the DMLEþ2.0 software program (www.dmle.org). Login to comment
47 Results Figure 2a shows the frequencies of the nine microsatellitic nucleotide repeats in chromosomes of DJS patients bearing the MRP2 I1173F mutation and normal chromosomes. Login to comment
49 The differences in the allele frequencies between normal and mutants were statistically significant (P < 0.05) for all microsatellites, which is consistent with a founder effect for the I1173F mutation. Login to comment
53 The DMLEþ2.0 program was then used for estimating the age of the MRP2 I1173F mutation and the FVII A244V mutation using the microsatellite data from patients and controls. Login to comment
54 Figure 3 shows the posterior probability densities of the mutation age for the MRP2 I1173F and FVII A244V. Login to comment
55 These mutation densities depict a peak at 75 generations for MRP2 I1173F and at 129 generations for FVII A244V. Login to comment
56 Assuming 20 years for a generation, the age of the MRP2 I1173F mutation was estimated to be 1500 years (95% credible set 660-3820) and the age of the FVII A244V mutation was estimated to be 2580 years (95% credible set 1440-4840). Login to comment
67 We also showed that DJS is prevalent in both populations and is caused by distinct mutations, I1173F in Iranian Jews and R1150H in Moroccan Jews [17]. Login to comment
69 We investigated this assumption by estimating the age of FVII A244V and MRP2 I1173F using the DMLEþ 2.0 program. Login to comment
71 The highest probability of the estimated age of FVII A244V was approximately 2600 years and that of MRP2 I1173F was approximately 1500 years. These age estimates had wide 95% credible sets, 660-3820 years for the MRP2 I1173F mutation and 1440-4840 years for the FVII A244V mutation, respectively. Login to comment

PMID: 16847695 [PubMed] Nies AT et al: "The apical conjugate efflux pump ABCC2 (MRP2)."

No. Sentence Comment

139 Although all sequence variants associated with Dubin-Johnson syndrome result in the absence of a Table 3 Nucleotide sequence variants in the human ABCC2 gene (NM_000392) leading to amino acid changes in the ABCC2/MRP2 protein (NP_000383) Location Nucleotide changea Deduced effect on proteina Causing Dubin-Johnson syndromeb Predicted effect by PolyPhen databasec Experimentally proven functional consequence Average frequency of indicated nucleotide exchange in population NCBI SNP IDd and/or references Exon 2 c.56 C>Te p.P19L Probably damaging T: 0.007 [63] Exon 2 c.116 T>A p.F39Y Benign A: 0.010 rs927344 A: 0.008 rs17222603 Exon 3 c.298 C>T p.R100Xf DJS [154] Exon 3 c.299 G>Ae p.R100Q Possibly damaging A: 0.007 [63] Exon 7 c.736 A>C p.M246L Benign C: 0.002 rs8187667 C: 0.002 rs17222744 Exon 7 c.842 G>A p.S281N Benign A: 0.0060.056 [117] Exon 8 c.998 A>G p.D333G Possibly damaging G: 0.002 rs8187668 G: 0.004 rs17222674 Exon 9 c.1058 G>A p.R353H Benign A: 0.009 rs7080681 A: 0.014 rs17216205 Exon 9 c.1177 C>T p.R393W DJS Probably damaging [104, 112] Exon 10 c.1234 A>G p.R412G Probably damaging Deficient methotrexate transport function [56] Exon 10 c.1249 G>A p.V417I Benign None apparent [50] A: 0.163 rs2273697, [146] A: 0.158 rs17216184 A: 0.125 [62] A: 0.1830.312 [117] Exon 10 c.1457 C>T p.T486I Benign T: 0.002 rs8187670 T: 0.002 rs17222589 Exon 11 c.1483 A>G p.K495E Possibly damaging G: 0.002 rs8187672 G: 0.002 rs17222561 Exon 13 c.1686 T>G p.F562L Benign G: 0.002 rs8187673 G: 0.002 rs17216233 Exon 16 c.2009 T>C p.I670T Benign rs8187676 C: 0.006 rs17222632 Exon 16 c.2026 G>C p.G676R DJS Probably damaging [181] Exon 17 c.2125 T>C p.W709R DJS Probably damaging [111] Exon 17 c.2153 A>G p.N718S Possibly damaging rs3740072 Exon 17 c.2215 C>T p.L739F Probably damaging T: 0.006 [51] Exon 18 c.2302 C>T p.R768W DJS Probably damaging Deficient maturation and impaired sorting [47] T: 0.010 [62] [168, 180] Exon 18 c.2366 C>T p.S789F Probably damaging Reduced protein levels [50] T: 0.010 [62] Exon 19 c.2546 T>G p.L849R Benign G: 0.002 rs8187689 G: 0.006 rs17222617 Exon 20 c.2647 G>Ae p.D883N Benign A: 0.007 [63] Exon 20 c.2677 G>C p.E893Q Benign rs3740071 Exon 21 c.2882 A>Ge p.K961R Benign G: 0.007 [63] Exon 22 c.2901 C>A p.Y967Xf A: 0.002 rs8187683 A: 0.002 rs17222547 Exon 22 c.2944 A>G p.I982V Benign G: 0.002 rs8187684 G: 0.002 rs17222554 Exon 22 c.3057 G>Te p.Q1019H Benign T: 0.007 [63] Exon 23 c.3107 T>C p.I1036T Possibly damaging C: 0.002 rs8187685 C: 0.004 rs17216149 Exon 23 c.3188 A>G p.N1063S Benign G: 0.002 rs8187686 G: 0.002 rs17222540 Exon 23 c.3196 C>T p.R1066Xf DJS No ABCC2 protein in liver [134] Exon 25 c.3449 G>A p.R1150H DJS Probably damaging Deficient transport function A: 00.009 [117] Exon 25 c.3517 A>T p.I1173F DJS Probably damaging Deficient maturation and impaired sorting, deficient transport function T: 00.029 [117] [80, 117] Exon 25 c.3521 G>Ae p.R1174H Probably damaging A: 0.007 [63] Exon 25 c.3542 G>T p.R1181L Possibly damaging T: 0.039 rs8187692 T: 0.034 rs17222702 Exon 25 c.3563 T>A p.V1188E Benign A: 0.059 rs8187694 A: 0.059 rs17222723 Exon 26 c.3732 T>Ge p.N1244K Possibly damaging G: 0.014 [63] Exon 27 c.3817 A>G p.T1273A Benign G: 0.002 rs8187699 G: 0.004 rs17222582 Exon 27 c.3825 C>G p.Y1275Xf DJS No ABCC2 protein in liver [104] Exon 28 c.3872 C>T p.P1291L Possibly damaging T: 0.012 rs8187700 T: 0.010 rs17216317 Exon 28 c.3895 A>C p.K1299Q Benign rs4148400, [146] Exon 28 c.3928 C>T p.R1310Xf DJS [166] Exon 29 c.4100 C>Ge p.S1367C Possibly damaging G: 0.007 [63] Exon 29 c.4145 A>G p.Q1382R DJS Probably Deficient [47, 168] Table 3 (continued) Location Nucleotide changea Deduced effect on proteina Causing Dubin-Johnson syndromeb Predicted effect by PolyPhen databasec Experimentally proven functional consequence Average frequency of indicated nucleotide exchange in population NCBI SNP IDd and/or references functionally active ABCC2 protein from the canalicular membrane, their effects on the synthesis and function of the ABCC2 protein differ. Login to comment
140 Although all sequence variants associated with Dubin-Johnson syndrome result in the absence of a Table 3 Nucleotide sequence variants in the human ABCC2 gene (NM_000392) leading to amino acid changes in the ABCC2/MRP2 protein (NP_000383) Location Nucleotide changea Deduced effect on proteina Causing Dubin-Johnson syndromeb Predicted effect by PolyPhen databasec Experimentally proven functional consequence Average frequency of indicated nucleotide exchange in population NCBI SNP IDd and/or references Exon 2 c.56 C>Te p.P19L Probably damaging T: 0.007 [63] Exon 2 c.116 T>A p.F39Y Benign A: 0.010 rs927344 A: 0.008 rs17222603 Exon 3 c.298 C>T p.R100Xf DJS [154] Exon 3 c.299 G>Ae p.R100Q Possibly damaging A: 0.007 [63] Exon 7 c.736 A>C p.M246L Benign C: 0.002 rs8187667 C: 0.002 rs17222744 Exon 7 c.842 G>A p.S281N Benign A: 0.0060.056 [117] Exon 8 c.998 A>G p.D333G Possibly damaging G: 0.002 rs8187668 G: 0.004 rs17222674 Exon 9 c.1058 G>A p.R353H Benign A: 0.009 rs7080681 A: 0.014 rs17216205 Exon 9 c.1177 C>T p.R393W DJS Probably damaging [104, 112] Exon 10 c.1234 A>G p.R412G Probably damaging Deficient methotrexate transport function [56] Exon 10 c.1249 G>A p.V417I Benign None apparent [50] A: 0.163 rs2273697, [146] A: 0.158 rs17216184 A: 0.125 [62] A: 0.1830.312 [117] Exon 10 c.1457 C>T p.T486I Benign T: 0.002 rs8187670 T: 0.002 rs17222589 Exon 11 c.1483 A>G p.K495E Possibly damaging G: 0.002 rs8187672 G: 0.002 rs17222561 Exon 13 c.1686 T>G p.F562L Benign G: 0.002 rs8187673 G: 0.002 rs17216233 Exon 16 c.2009 T>C p.I670T Benign rs8187676 C: 0.006 rs17222632 Exon 16 c.2026 G>C p.G676R DJS Probably damaging [181] Exon 17 c.2125 T>C p.W709R DJS Probably damaging [111] Exon 17 c.2153 A>G p.N718S Possibly damaging rs3740072 Exon 17 c.2215 C>T p.L739F Probably damaging T: 0.006 [51] Exon 18 c.2302 C>T p.R768W DJS Probably damaging Deficient maturation and impaired sorting [47] T: 0.010 [62] [168, 180] Exon 18 c.2366 C>T p.S789F Probably damaging Reduced protein levels [50] T: 0.010 [62] Exon 19 c.2546 T>G p.L849R Benign G: 0.002 rs8187689 G: 0.006 rs17222617 Exon 20 c.2647 G>Ae p.D883N Benign A: 0.007 [63] Exon 20 c.2677 G>C p.E893Q Benign rs3740071 Exon 21 c.2882 A>Ge p.K961R Benign G: 0.007 [63] Exon 22 c.2901 C>A p.Y967Xf A: 0.002 rs8187683 A: 0.002 rs17222547 Exon 22 c.2944 A>G p.I982V Benign G: 0.002 rs8187684 G: 0.002 rs17222554 Exon 22 c.3057 G>Te p.Q1019H Benign T: 0.007 [63] Exon 23 c.3107 T>C p.I1036T Possibly damaging C: 0.002 rs8187685 C: 0.004 rs17216149 Exon 23 c.3188 A>G p.N1063S Benign G: 0.002 rs8187686 G: 0.002 rs17222540 Exon 23 c.3196 C>T p.R1066Xf DJS No ABCC2 protein in liver [134] Exon 25 c.3449 G>A p.R1150H DJS Probably damaging Deficient transport function A: 00.009 [117] Exon 25 c.3517 A>T p.I1173F DJS Probably damaging Deficient maturation and impaired sorting, deficient transport function T: 00.029 [117] [80, 117] Exon 25 c.3521 G>Ae p.R1174H Probably damaging A: 0.007 [63] Exon 25 c.3542 G>T p.R1181L Possibly damaging T: 0.039 rs8187692 T: 0.034 rs17222702 Exon 25 c.3563 T>A p.V1188E Benign A: 0.059 rs8187694 A: 0.059 rs17222723 Exon 26 c.3732 T>Ge p.N1244K Possibly damaging G: 0.014 [63] Exon 27 c.3817 A>G p.T1273A Benign G: 0.002 rs8187699 G: 0.004 rs17222582 Exon 27 c.3825 C>G p.Y1275Xf DJS No ABCC2 protein in liver [104] Exon 28 c.3872 C>T p.P1291L Possibly damaging T: 0.012 rs8187700 T: 0.010 rs17216317 Exon 28 c.3895 A>C p.K1299Q Benign rs4148400, [146] Exon 28 c.3928 C>T p.R1310Xf DJS [166] Exon 29 c.4100 C>Ge p.S1367C Possibly damaging G: 0.007 [63] Exon 29 c.4145 A>G p.Q1382R DJS Probably Deficient [47, 168] Table 3 (continued) Location Nucleotide changea Deduced effect on proteina Causing Dubin-Johnson syndromeb Predicted effect by PolyPhen databasec Experimentally proven functional consequence Average frequency of indicated nucleotide exchange in population NCBI SNP IDd and/or references functionally active ABCC2 protein from the canalicular membrane, their effects on the synthesis and function of the ABCC2 protein differ. Login to comment

PMID: 16952291 [PubMed] Corpechot C et al: "Identification of a novel 974C-->G nonsense mutation of the MRP2/ABCC2 gene in a patient with Dubin-Johnson syndrome and analysis of the effects of rifampicin and ursodeoxycholic acid on serum bilirubin and bile acids."

No. Sentence Comment

78 Mutations in the MRP2/ABCC2 Gene Associated with DJS Nucleotide Mutation Exon Predicted Effect Reference 298C→T 3 R100X 27 974C→G 8 S325X This article IVS8 + 4A→G Intron 8 Aberrant splicing 28 1177C→T 9 R393W 29 1256insCT/ delAAACAG TGAACCT- GATG 10 Frameshift 30 1271A→G 10 R412G 31 1815 + 2T→A 13 Skipped exon 32, 33 1967 + 2T→C 15 Skipped exon 34, 35 2026G→C 16 G676R 35 2125T→C 17 W709R 36 2302C→T 18 R768W 32, 37, 38 2439 + 2T→C 18 Skipped exon 32, 35, 37 3196C→T 23 R1066X 39, 40 3449G→A 25 R1150H 41 3517A!92;T 25 I1173F 41 3928C→T 28 R1310X 27, 33 4145A→G 29 Q1382R 37 4175delGGATGA 30 R1392 + M1393 deletion 40 4292delCA 30 Frameshift 30 DISCUSSION Identification of a Novel Nonsense Mutation of the MRP2/ABCC2 Gene Up to now, 18 mutations in the sequence of the MRP2/ABCC2 gene have been reported in DJS, including nonsense mutations, deletions, splicing junction mutations, and missense mutations (Table 1). Login to comment

PMID: 16377077 [PubMed] Wada M et al: "Single nucleotide polymorphisms in ABCC2 and ABCB1 genes and their clinical impact in physiology and drug response."

No. Sentence Comment

41 In Japan, the expected number of Table 1 Summary of mutations identified in Dubin-Johnson syndrome (DJS) Mutation Exon IVS Amino acid alteration Reference 298COT 3 R100X a,b 1815C 2TOA 13 Exon13 skip [38] 1967C 2TOC 15 Exon15 skip [62] 2026GOC 16 G676R [92] 2302COT 18 R768W [49,91]c 2439C 2TOC 18 Exon18 skip [38]a,c 3196COT 23 R1066X [47] 3449GOA 25 R1150H [52] 3517AOT 25 I1173F [52] 3928COT 28 R1310X [50] 4145AOG 29 Q1382R [38] 4175- 4180del 30 RM1392-1393del [48] a Adachi and Wada, unpublished data. b Houkibara and Wada, unpublished data. Login to comment
59 [42,49,53,91]c 18 2366COT S789F NBD1 (Transport activity) Not reported 0.9 [42]a 25 3449GOA R1150H MSD3 DJS (transport activity) 0.3 Not reported [52] 25 3517AOT I1173F MSD3 DJS (protein maturation) 1.4 Not reported [52] 28 3895AOC K1299Q NBD2 Unkown Not reported 1? Login to comment
67 In contrast, expression of another mutant ABCC2 (I1173F) was low and mislocated to the ER of the transfected cells. Login to comment

PMID: 16180115 [PubMed] Ito K et al: "Apical/basolateral surface expression of drug transporters and its role in vectorial drug transport."

No. Sentence Comment

236 At least three of the other mutations (R768W, I1173F, and deletion 1392Y1394) are related to sorting problems from the ER to Golgi, as these mutant MRP2 accumulated within the ER in core-glycosylated forms. Login to comment

PMID: 16012956 [PubMed] Cebecauerova D et al: "Dual hereditary jaundice: simultaneous occurrence of mutations causing Gilbert's and Dubin-Johnson syndrome."

No. Sentence Comment

61 3517AϾT 25 I1173F Mor-Cohen R et al, J Biol Chem 2001;276:36923-36930. Login to comment

PMID: 14699511 [PubMed] Kullak-Ublick GA et al: "Enterohepatic bile salt transporters in normal physiology and liver disease."

No. Sentence Comment

131 The syndrome is caused by the absence of MRP2 protein from the canalicular hepatocyte membrane182 because of mutations of the MRP2 gene (ABCC2).181,183,184 The MRP2Delta(R,M) mutation, which describes the deletion of Arg1392 and Met1393, causes disturbed maturation and trafficking of the protein from the endoplasmic reticulum to the Golgi complex and impaired sorting of the glycoprotein to the apical membrane.185 The MRP2 I1173F mutation, which denotes the exchange of the hydrophobic amino acid isoleucine 1173 with phenylalanine in a predicted extracellular loop of MRP2, leads to retention of MRP2 in the endoplasmic reticulum and degradation by proteasomes in transfected HEK293 cells, and is associated with a loss of ATP-dependent transport of leukotriene C4.186 Absent MRP2 function may be compensated for by increased expression of MRP3 at the basolateral hepatocyte membrane, as suggested by immunofluorescence studies on liver sections from a Dubin-Johnson patient.61 Polymorphisms of basolateral bile salt transporter genes. Login to comment

PMID: 12388192 [PubMed] Keitel V et al: "A common Dubin-Johnson syndrome mutation impairs protein maturation and transport activity of MRP2 (ABCC2)."

No. Sentence Comment

8 In this work, we analyzed a relatively frequent Dubin-Johnson syndrome mutation that leads to an exchange of two hydrophobic amino acids, isoleucine 1173 to phenylalanine (MRP2I1173F), in a predicted extracellular loop of MRP2. Login to comment

PMID: 12884082 [PubMed] Wakusawa S et al: "Identification of a novel 2026G-->C mutation of the MRP2 gene in a Japanese patient with Dubin-Johnson syndrome."

No. Sentence Comment

43 With respect to the polymorphism )24C fi T, we also employed a restriction enzyme assay with BbsI; the results showed that his spouse and one daughter were heterozygous for this SNP, but other Table 1 Mutations of MRP2 in Dubin-Johnson syndrome (DJS) Nucleotide mutation Predicted effects References Splice site mutation 1815+2T fi A 1669del147 (exon13 skipping) Wada et al. 1998 1967+2T fi C 1901del67 (exon15 skipping) Kajihara et al. 1998 2439+2T fi C 2272del168 (exon18 skipping) Toh et al. 1999 Deletion mutation Del4170-5 Del R1392 , M1393 Tsujii et al. 1999 Missense mutation 2302C fi T R768 W Wada et al. 1998 3449G fi A R1150H Mor-Cohen et al. 2001 3517A fi T I1173F Mor-Cohen et al. 2001 4145A fi G Q1382R Toh et al. 1999 Nonesense mutation 3196C fi T R1066X Paulusma et al. 1997 3928C fi T R1310X Tate et al. 2002 family members did not possess it (Fig. 4). Login to comment

PMID: 12406647 [PubMed] Suzuki H et al: "Single nucleotide polymorphisms in multidrug resistance associated protein 2 (MRP2/ABCC2): its impact on drug disposition."

No. Sentence Comment

109 mutation, and deletion mutation have all been re- Moreover, I1173F and R1150H have been found ported. Login to comment

PMID: 12130697 [PubMed] Gerk PM et al: "Regulation of expression of the multidrug resistance-associated protein 2 (MRP2) and its role in drug disposition."

No. Sentence Comment

53 Site-directed mutagenesis studies substi- TABLE 1 Mutations in human MRP2 Mutation Location Translation Domain Phenotype Reference -24(C-T) Promoter 5Ј-UTR Not reported (Ito et al., 2001e) 1249(G-A)/wt Exon 10 V4171 MSD2 Not reported (Ito et al., 2001e) 1815ϩ2(T-A)/1815ϩ2(T-A) Exon 13 147-bp deletion MSD2 DJS (Wada et al., 1998; Toh et al., 1999) 2002del67/2002del67 Exon 16 Premature termination codon NBD1 DJS (Konig et al., 1999a) 2302(C-T)/2302(C-T) Exon 18 R768W/R768W NBD1 DJS (Wada et al., 1998; Toh et al., 1999; Ito et al., 2001e) 2302(C-T)/2439ϩ2(T-C) Exon 18 R768W/168-bp deletion NBD1 DJS (Toh et al., 1999) 2302(C-T)/wt Exon 18 R768W/wt NBD1 Increased UCP1 (Toh et al., 1999) 2366(C-T)/wt Exon 18 S789F/wt NBD1 Not reported (Ito et al., 2001e) 2439ϩ2(T-C)/2439ϩ2(T-C) Exon 18 168-bp deletion/168-bp deletion NBD1 DJS (Wada et al., 1998; Toh et al., 1999) 2439ϩ2(T-C)/4145(A-G) Exon 18/29 168-bp deletion/Q1328R NBD1/2 DJS (Toh et al., 1999) 2439ϩ2(T-C)/wt Exon 18 168-bp deletion/wt NBD1 Increased UCP1 (Toh et al., 1999) 3196(C-T)/3196(C-T) Exon 23 Premature termination codon MSD3 DJS (Paulusma et al., 1997; Tsujii et al., 1999) 3449(G-A)/3449(G-A) Exon 25 R1150H/R1150H MSD3 DJS (Mor-Cohen et al., 2001) 3517(A-T)/3517(A-T) Exon 25 I1173F/I1173F MSD3 DJS (Mor-Cohen et al., 2001) 3972(C-T)/wt Exon 28 I1324I/wt near NBD2 None (Ito et al., 2001e) 4175del6 Exon 30 Loss of R1392 and M1393 NBD2 DJS (Tsujii et al., 1999) 4348(G-A)/wt Exon 31 A1450T/wt NBD2 Not reported (Ito et al., 2001e) UTR, untranslated region; wt, wild type; bp, base pair; UCP1, urinary coproporphyrin fraction 1. tuting the cationic amino acid Arg586 and Arg1096 in rat Mrp2 with neutral amino acids (R586L, R586I, R1096L, and R1096M) or a cationic amino acid (R1096K) led to acquisition of taurocholate transport and retention of glucuronide and glutathione conjugate transport by Mrp2 (Ito et al., 2001d). Login to comment

PMID: 11477083 [PubMed] Mor-Cohen R et al: "Identification and functional analysis of two novel mutations in the multidrug resistance protein 2 gene in Israeli patients with Dubin-Johnson syndrome."

No. Sentence Comment

3 One mutation, 3517A3T, predicting a I1173F substitution, was found in 22 homozygous Iranian Jewish DJS patients from 13 unrelated families and a second mutation, 3449G3A, predicting a R1150H substitution, was found in 5 homozygous Moroccan Jewish DJS patients from 4 unrelated families. Login to comment
8 In contrast, expression of MRP2 (I1173F) was low and mislocated to the endoplasmic reticulum of the transfected cells. These findings provide an explanation for the DJS phenotype in these two patient groups. Login to comment
63 Incorporation of the mutations was verified by DNA sequencing. Two sets of primers were used to introduce the I1173F mutation:1) the forward primer (5Ј-CAGGTTT- GCCAGTTTTCCGTGCCTTTGAGC-3Ј) and the reverse primer (5Ј- GCTCAAAGGCACGGAAAACTGGCAAACCTG-3Ј); 2) the forward primer (5Ј-CCGTATCAGGTTTGCCAGTTTTCCGTGCCTTTGAGC-3Ј) and the reverse primer (5Ј-GCTCAAAGGCACGGAAAACTGGCAAAC- CTGATACGG-3Ј). Login to comment
70 Exon Fragment size Forward primer Reverse primer Name Sequence Name Sequence base pairs 1 404 1F 5Ј-TTGTTGGCCAGCTCTGTTG-3Ј 1R* 5Ј-ACTACCACTTGTTCTGAGTC-3Ј 2 310 2F* 5Ј-TGAAAGCAGTGGGATGTGC-3Ј 2R* 5Ј-CTCTACTGTGCAGCCAAGG-3Ј 3 295 3F* 5Ј-ATCTGAATCACTGCATACCG-3Ј 3R* 5Ј-TCACCTAGATGCCTATGGG-3Ј 4 251 4F* 5Ј-CTCAGTCCTCGGTTAGTGG-3Ј 4R* 5Ј-CTATGAGTTAGAGGTTGCCC-3Ј 5 266 5F* 5Ј-GCCATGTAGACTTCCTTTGG-3Ј 5R* 5Ј-ACCTTATTCTGGGCTTGTGG-3Ј 6 185 6F* 5Ј-TTAGAGTCCCATGAAGTTCC-3Ј 6R* 5Ј-AGTAAGGATACAGCCAATCC-3Ј 7 406 7F* 5Ј-TGGAGATAGCCTCTGACCC-3Ј 7R 5Ј-TGCACTGAGAAGTATGAAGTGC-3Ј 8 428 8F 5Ј-CCTGTACAGAGAAGGCCACG-3Ј 8R 5Ј-CGGTCTTCATGACACAATGC-3Ј 9 515 9F* 5Ј-GATAGTGTAGTCTAGCTGGC-3Ј 9R* 5Ј-TGAGCACCAGAACAGCTTGC-3Ј 10 435 10F* 5Ј-ACTCCCTAGTATCCTTGGC-3Ј 10R* 5Ј-GATGGTAGAAAGTCTTCCACCAGC-3Ј 11 348 11F 5Ј-ACAGTCAGGCAAGGGCTATG-3Ј 11R 5Ј-TCCTTACCCACAGAGAGCC-3Ј 12 389 12F* 5Ј-GGATCAGATACACCTGGTGC-3Ј 12R* 5Ј-ACGAAGGTGAAACTAGAGC-3Ј 13 512 13F* 5Ј-AAGGATTGGCTTAGGAGGC-3Ј 13R* 5Ј-AGTCATTCTGGACTCCAAGG-3Ј 14 256 14F* 5Ј-TTAGGAGATGCCAGCTGTGG-3Ј 14R* 5Ј-ATTCTGGCACCAGTACTGCG-3Ј 15 286 15F* 5Ј-GCACTTAGCAGAAACAATCC-3Ј 15R* 5Ј-ACCGAAGACATGCACATAGC-3Ј 16 343 16F* 5Ј-CCTGATACCAGACTTCATGG-3Ј 16R* 5Ј-GTCGGATGTCTCCAAGACC-3Ј 17 289 17F* 5Ј-CTTCAACCCTGCGTTTCTGG-3Ј 17R* 5Ј-CTCTTCAATATGCCTTCACCC-3Ј 18 ϩ 19 792 18F 5Ј-TCACAGGGTGACAAGCAAC-3Ј 19R 5Ј-TTTACCATTCCACCCATGGC-3Ј 20 352 20F 5Ј-GTGTCTCCCTAGTCCATGATGG-3Ј 20R 5Ј-TCACTCAGCTGGCATCAAAG-3Ј 21 330 21F* 5Ј-ATGCGCTTTGATGCCAGCTG-3Ј 21R* 5Ј-ATTGCTCCTGTAAGTATGCG-3Ј 22 417 22F* 5Ј-TTGGCATTCTAGGTGATTCC-3Ј 22R* 5Ј-CACCATGCACAGGAATCCC-3Ј 23 352 23F* 5Ј-CACAAGTCTTCAGGGATTCC-3Ј 23R* 5Ј-GGTACTCAAGAAACACTTGC-3Ј 24 307 24F* 5Ј-TTACATGAAGGAGTACTGGG-3Ј 24R* 5Ј-GGAAGGATGACTTAGCAATTTCC-3Ј 25 460 25F 5Ј-GGAGCCTCTCATCATTCTGC-3Ј 25R 5Ј-TTTCACACCACTAGCCATGC-3Ј 26 402 26F 5Ј-GAGGCATTGCCTAAGAGTGC-3Ј 26R 5Ј-AAAGATGGAGCCAGGGTTTG-3Ј 27 214 27F 5Ј-TTGGTTTCTGTGCCTATGATG-3Ј 27R* 5Ј-GCACTCTCGAAGGAGTTGC-3Ј 28 323 28F* 5Ј-TCTATGTCTCGAGTCCTGGG-3Ј 28R* 5Ј-CAAATGATGAAGGCTTAGGG-3Ј 29 285 29F* 5Ј-ATGGAGTAGCCAGTCACTGC-3Ј 29R* 5Ј-CCCGAGTAAGTTCTAGAGC-3Ј 30 321 30F* 5Ј-CAGGAATCCATCTCAGGCC-3Ј 30R* 5Ј-CACATCCTCTCATTGCCTGC-3Ј 31 282 31F* 5Ј-CTTTAGGAGCTAACACATGG-3Ј 31R* 5Ј-GAGCAAGGGTTAAGCCATCC-3Ј 32 192 32F* 5Ј-AATGCCTAGACTTGAGATGC-3Ј 32R 5Ј-CTGCTAGAATTTTGTGCTGTTCACATTC-3Ј three clones of each mutant were analyzed for activity, and all six clones of the I1173F mutant were analyzed for expression by Western blot. Login to comment
97 RESULTS Identification of Candidate Mutations in DJS Patients- Screening of all 32 exons of the MRP2 gene in an Iranian Jewish patient disclosed a 3517A3T transition in exon 25, predicting an I1173F substitution. Login to comment
157 Carboxyfluorescein Transport of Mutants MRP2-Transfected HEK-293 cells expressing the I1173F- and R1150H-MRP2 mutants were used to study the effect of the mutations on CF transport. Login to comment
160 Cells transfected with either I1173F-MRP2 (Fig. 5C) or R1150H-MRP2 (Fig. 5D) showed a slow efflux of CF, which was similar to that measured in control cells (Figs. Login to comment
164 The average rate constant of CF efflux (in s-1 ) by WT-MRP2 (562 Ϯ 56, n ϭ 4) was significantly higher than that measured in control cells (151 Ϯ 45, n ϭ 5), R1150H-MRP2 cells (90 Ϯ 40, n ϭ 5) and I1173F-MRP2 cells (104 Ϯ 66, n ϭ 3). Login to comment
165 No significant difference was found between the average rate constant of control, R1150H-MRP2, and I1173F-MRP2 cells. These data indicate TABLE III Frequency of haplotypes in Iranian Jewish and Moroccan Jewish controls and DJS patients Haplotype 5Ј-Untranslated region -24C3T Exon 7 842G3A Exon 10 1249G3A IVS29-35 G3A Frequency in Iranian Jewsa Frequency in Moroccan Jewsa Controls (n ϭ 144) Patients (n ϭ 26) Controls (n ϭ 118) Patients (n ϭ 8) % % 1 C G A A 0 100 0 0 2 C A G G 0.7 0 1.7 100 3 C G G G 63.2 0 61 0 4 C G A G 23.6 0 14.4 0 5 T G G G 11.1 0 18.6 0 Seven additional haplotypes 2.1 0 4.3 0 a n indicates number of informative alleles of unrelated individuals. Login to comment
178 Panel E shows the irreversible inhibition of efflux by cyclosporine A. that both I1173F and R1150H mutations impair the transport activity of MRP2. Login to comment
187 By contrast, the amount of protein and the ratio between the two bands was different for I1173F-MRP2. Login to comment
188 At 20 ␮g of I1173F-MRP2 protein, staining was much weaker than that observed with either WT or R1150H-MRP2. Login to comment
189 When 55 ␮g of protein from cells expressing I1173F-MRP2 was used, the overall amount of protein was comparable to that measured with 13 ␮g of WT or R1150H-MRP2. Login to comment
190 Notably, the 175-kDa band was more intense than the 190-kDa band for I1173F-MRP2. Login to comment
191 Cellular Localization of WT and Mutants MRP2-The significant amount of the 175-kDa band found in cells expressing I1173F-MRP2 suggests that processing of MRP2 was affected by this mutation. Login to comment
196 In contrast, I1173F-MRP2 showed largely a reticular pattern of staining (Fig. 7C). Login to comment
199 Both mutations are located in exon 25 of the MRP2 gene, with 3517A3T predicting I1173F substitution found in a cluster of Iranian Jewish DJS patients, and 3449G3A predicting R1150H substitution found in a cluster of Moroccan Jewish patients. Login to comment
218 Cells were transfected with GFP only (control, A), WT-MRP2 (B), I1173F-MRP2 (C), or R1150H-MRP2 (D). Login to comment
233 With this assay we showed that both I1173F and R1150H mutations impaired the MRP2 activity (Fig. 5). Login to comment
238 In contrast, the I1173F mutation gave rise to a markedly reduced relative amount of the 190-kDa band and an increased relative amount of the 175-kDa form. Login to comment
239 Furthermore, the total amount of I1173F-MRP2 that was expressed was substantially decreased (Fig. 6), which suggests enhanced intracellular degradation, due to mistargeting and misfolding of the mutant protein. Login to comment
240 Indeed, immunolocalization of the different constructs showed that WT and R1150H-MRP2 were properly targeted to the plasma membrane, whereas I1173F-MRP2 remained largely confined to the ER. Login to comment
242 Our expression studies suggest that the I1173F mutation causes impaired maturation of MRP2, mislocalization probably to the ER, and augmented degradation, whereas the R1150H mutation does not affect the maturation and localization, but impairs the MRP2 transport activity. Login to comment
252 HEK 293 cells transfected with WT-MRP2 (A), R1150H-MRP2 (B), or I1173F-MRP2 (C) were detected by the monoclonal antibody M2III-6 and were examined by confocal laser scanning microscopy. Login to comment

PMID: 16815813 [PubMed] Choudhuri S et al: "Structure, function, expression, genomic organization, and single nucleotide polymorphisms of human ABCB1 (MDR1), ABCC (MRP), and ABCG2 (BCRP) efflux transporters."

No. Sentence Comment

340 Additional missense mutations Ile1173Phe and Arg1150His have been found in Iranian Jewish and Moroccan Jewish DJS patients, respectively. Login to comment