ABCMdb

ABCA1 p.Arg282*

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[hide] Increased risk of coronary artery disease in Cauca... Biochim Biophys Acta. 2012 Mar;1821(3):416-24. doi: 10.1016/j.bbalip.2011.08.006. Epub 2011 Aug 19. Tietjen I, Hovingh GK, Singaraja R, Radomski C, McEwen J, Chan E, Mattice M, Legendre A, Kastelein JJ, Hayden MR
Increased risk of coronary artery disease in Caucasians with extremely low HDL cholesterol due to mutations in ABCA1, APOA1, and LCAT.
Biochim Biophys Acta. 2012 Mar;1821(3):416-24. doi: 10.1016/j.bbalip.2011.08.006. Epub 2011 Aug 19., [PMID:21875686]

Abstract [show]
Mutations in ABCA1, APOA1, and LCAT reduce HDL cholesterol (HDLc) in humans. However, the prevalence of these mutations and their relative effects on HDLc reduction and risk of coronary artery disease (CAD) are less clear. Here we searched for ABCA1, APOA1, and LCAT mutations in 178 unrelated probands with HDLc <10th percentile but no other major lipid abnormalities, including 89 with >/=1 first-degree relative with low HDLc (familial probands) and 89 where familial status of low HDLc is uncertain (unknown probands). Mutations were most frequent in LCAT (15.7%), followed by ABCA1 (9.0%) and APOA1 (4.5%), and were found in 42.7% of familial but only 14.6% of unknown probands (p=2.44 *10(-5)). Interestingly, only 16 of 24 (66.7%) mutations assessed in families conferred an average HDLc <10th percentile. Furthermore, only mutation carriers with HDLc <5th percentile had elevated risk of CAD (odds ratio (OR)=2.26 for 34 ABCA1 mutation carriers vs. 149 total first-degree relative controls, p=0.05; OR=2.50 for 26 APOA1 mutation carriers, p=0.04; OR=3.44 for 38 LCAT mutation carriers, p=1.1 *10(-3)). These observations show that mutations in ABCA1, APOA1, and LCAT are sufficient to explain >40% of familial hypoalphalipoproteinemia in this cohort. Moreover, individuals with mutations and large reductions in HDLc have increased risk of CAD. This article is part of a Special Issue entitled Advances in High Density Lipoprotein Formation and Metabolism: A Tribute to John F. Oram (1945-2010).

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117 COOHA B T929I H2N R587W B A M1091T C1477R K776N N935S S1181F IVS24+1 G>C V2244I R282X D571G M640L S930F M968T R1615WIVS16-5 CA>del ABCA1 Transmembrane domain ATP-binding domain Q597R A) AA 1 AA 267 K130del L202P 74 90 98 112 122 145 167 189 211 215 233 253 APOA1 Negative charge domain Alpha-helix E222K E134DT37M 140 178 206 41127 104 121 165 200 229 360 391 Y135N V246F 127 206 369 401 Catalytic triad R322C L338H V371MV52M Y107X A117T T147I V333M Phe Leu Asp His AA 1 AA 440 R159Q I202T LCAT Alpha helixBeta sheet B) 419I.Tietjenetal./BiochimicaetBiophysicaActa1821(2012)416-424 3.4. Login to comment

[hide] Identification and characterization of novel loss ... Atherosclerosis. 2010 Dec;213(2):492-8. Epub 2010 Aug 26. Candini C, Schimmel AW, Peter J, Bochem AE, Holleboom AG, Vergeer M, Dullaart RP, Dallinga-Thie GM, Hovingh GK, Khoo KL, Fasano T, Bocchi L, Calandra S, Kuivenhoven JA, Motazacker MM
Identification and characterization of novel loss of function mutations in ATP-binding cassette transporter A1 in patients with low plasma high-density lipoprotein cholesterol.
Atherosclerosis. 2010 Dec;213(2):492-8. Epub 2010 Aug 26., [PMID:20880529]

Abstract [show]
OBJECTIVES: The current literature provides little information on the frequency of mutations in the ATP-binding cassette transporter A1 (ABCA1) in patients with low high-density lipoprotein cholesterol (HDL) levels that are referred to the clinic. In 78 patients with low plasma levels of HDL cholesterol that were referred to our clinic, we routinely screened for ABCA1 gene mutations and studied the functionality of newly identified ABCA1 missense mutations. METHODS: The coding regions and exon-intron boundaries of the ABCA1 gene were sequenced in 78 subjects with HDL cholesterol levels below the 10th percentile for age and gender. Novel mutations were studied by assessing cholesterol efflux capacity (using apolipoprotein A-I as acceptor) after transient expression of ABCA1 variants in BHK cells. RESULTS: Sixteen out of 78 patients (21%) were found to carry 19 different ABCA1 gene variants (1 frameshift, 2 splice-site, 4 nonsense and 12 missense variation) of which 14 variations were novel. Of three patients with homozygous mutations and three patients having compound heterozygous mutations only one patient presented with the clinical characteristics of Tangier Disease (TD) in the presence of nearly complete HDL deficiency. Seven out of eight newly identified ABCA1 missense mutations were found to exhibit a statistically significant loss of cholesterol efflux capacity. CONCLUSION: This study shows that one out of five patients who are referred to our hospital because of low HDL cholesterol levels have a functional ABCA1 gene mutation. It is furthermore demonstrated that in vitro studies are needed to assess functionality of ABCA1 missense mutations.

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76 Patients (gender, age) Amino acida (nucleotidea ) change TC TG LDL-c HDL-c Clinical manifestations of TD CVD Other relevant clinical data Homozygotes Patient 1 (female, 42) p.L1056P (c.3167T > C) 2.4 0.9 1.99 <0.10 Absent CAD Thrombocytopenia Patient 2 (male, 40) p.Wl747X (c.5240G > A) 1.76 1.93 0.52 0.1-0.3 Neuropathy, splenomegaly, thrombocytopenia Mild stenosis (20-30%) of coronary arteries None Patient 3 (male, 55) p.F593L (c.1779C > G) 4.4 1.4 3.6 <0.10 Absent CAD None p.E1253K (c.3757G > A) Compound heterozygotes Patient 4 (female, 63) p.Q1038X (c.3112C > T) 6.68 2.72 5.4 <0.10 Absent None None p.N1800H (c.5398A > C) [32] Patient 5 (female, 28) p.T1512M (c.4535C > T) 4.42 1.83 3.46 0.1 Absent None None p.N1800H (c.5398A > C) [32] p.C978fsX988 (c.2934delT) Patient 6 (female, 17) p.D575G (c.1724A > G) 4.96 2.84 4.35 <0.10 Absent None DM1 p.C1941R(c.5821T > C) Heterozygotes Patient 7 (male, 42) p.S100C (c.299C > G) 8.5 8.7 4.3 0.3 N.A. None None Patient 8 (male, 58) p.E1172D (c.3516G > C) [33] 6.4 2.7 4.1 0.9 N.A. None None Patient 9 (male, 35) p.S1181F (c.3542C > T) [17] 2.9 0.31 1.88 0.88 N.A. None None Patient 10 (male, 48) p.C1477R (c.4429T > C) [13] 2.01 1.4 0.92 0.46 N.A. CAD None Patient 11 (male, 68) p.V1858A (c.5573T > C) 4.9 3.78 2.41 0.75 N.A. CAD None Patient 12 (female, 36) p.N1800H (c.5398A > C) [32] 4.6 1.2 4 <0.10 N.A. None DM2, obesity Patient 13 (male, 67) p.R282X (c.844C > T) [34] 3.2 1.21 2.14 0.51 N.A. None DM2 Patient 14 (female, 42) p.W424X (c.1272G > A) 2.07 1.04 1.39 0.21 N.A. None None Patient 15 (female, 52) N.A. - (IVS11 - 1G > A) 5.51 3.51 3.28 0.56 N.A. None Hypothyroidism, hypertension Patient 16 (female, 54) N.A. - (IVS48 + 2T > C) 3.29 1.92 1.94 0.49 N.A. None DM2, hypertension a Nomenclature based on guidelines of Human Genome Variation Society. Login to comment
89 In ABCA1, we identified 14 novel and 5 known genetic variations in 16 subjects including one frameshift (p.C978fsX988), 2 splice-site (IVS11-1G > C and IVS48 + 2T > C), 4 nonsense (p.R282X, p.W424X, p.Q1038X, p.Wl747X) and 12 missense variations (p.S100C, p.D575G, p.F593L, p.L1056P, p.E1172D, p.S1181F, p.E1253K, p.C1477R, p.T1512M, p.N1800H, p.V1858A, p.C1941R). Login to comment

[hide] Tangier disease caused by compound heterozygosity ... Atherosclerosis. 2010 Mar;209(1):163-6. Epub 2009 Aug 29. Cameron J, Ranheim T, Halvorsen B, Kulseth MA, Leren TP, Berge KE
Tangier disease caused by compound heterozygosity for ABCA1 mutations R282X and Y1532C.
Atherosclerosis. 2010 Mar;209(1):163-6. Epub 2009 Aug 29., [PMID:19765707]

Abstract [show]
BACKGROUND: Inherited low levels of high density lipoprotein (HDL) cholesterol may be due to mutations in the genes encoding the ATP-binding cassette transporter A1 (ABCA1), apolipoprotein (apo) A-I or lecithin:cholesterol acyltransferase (LCAT). METHODS: The ABCA1, apoA-I and LCAT genes of a 40-year-old male subject with serum HDL cholesterol of 0.06mmol/l were subjected to DNA sequencing. The proband's family was examined for co-segregation between mutations and levels of HDL cholesterol. Cholesterol efflux in fibroblasts from the proband and a normocholesterolemic subject was compared. The effects of an ABCA1 mutation on cholesterol efflux and membrane localization of ABCA1 were studied in transfected HEK293 and HeLa cells, respectively. RESULTS: The proband was a compound heterozygote for ABCA1 mutations R282X (c.844 C>T) and Y1532C (c.4595 A>G). Relatives who were heterozygous for one of these mutations, had about half-normal HDL cholesterol levels. Cholesterol efflux was reduced in fibroblasts from the proband, as was cholesterol efflux from HEK293 cells transfected with an human (h) ABCA1 expression plasmid harboring the Y1532C mutation. Confocal microscopy of HeLa cells transfected with the Y1532C-hABCA1 plasmid revealed that the Y1532C mutation inhibits ABCA1 from reaching the cellular membrane. CONCLUSION: Compound heterozygosity for the nonsense mutation R282X and the missense mutation Y1532C in the ABCA1 gene causes Tangier disease. R282X has a detrimental effect on the function of ABCA1 since a premature stop codon is introduced. Mutation Y1532C disrupts the normal function of ABCA1 as determined by in vitro analyses.

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0 Atherosclerosis 209 (2010) -166 Contents lists available at ScienceDirect Atherosclerosis journal homepage: www.elsevier.com/locate/atherosclerosis Tangier disease caused by compound heterozygosity for ABCA1 mutations R282X and Y1532C Jamie Camerona , Trine Ranheima , Bente Halvorsenb,c , Mari Ann Kulsetha , Trond P. Lerena , Knut Erik Bergea,* a Medical Genetics Laboratory, Department of Medical Genetics, Rikshospitalet, Oslo University Hospital, NO-0027 Oslo, Norway b Research Institute of Internal Medicine, Rikshospitalet, Oslo University Hospital, Norway c University of Oslo, Norway a r t i c l e i n f o Article history: Received 10 July 2009 Received in revised form 18 August 2009 Accepted 19 August 2009 Available online 29 August 2009 Keywords: ABCA1 gene Cholesterol efflux HDL cholesterol Mutation Tangier disease a b s t r a c t Background: Inherited low levels of high density lipoprotein (HDL) cholesterol may be due to mutations in the genes encoding the ATP-binding cassette transporter A1 (ABCA1), apolipoprotein (apo) A-I or lecithin:cholesterol acyltransferase (LCAT). Login to comment
5 Results: The proband was a compound heterozygote for ABCA1 mutations R282X (c.844 C>T) and Y1532C (c.4595 A>G). Login to comment
9 Conclusion: Compound heterozygosity for the nonsense mutation R282X and the missense mutation Y1532C in the ABCA1 gene causes Tangier disease. Login to comment
10 R282X has a detrimental effect on the function of ABCA1 since a premature stop codon is introduced. Login to comment
66 However, DNA sequencing of the ABCA1 gene suggested that the proband was a compound heterozygote for the nonsense mutation R282X (c.844 C>T) in exon 9 and the missense mutation Y1532C (c.4595 A>G) in exon 34. Login to comment
67 R282X has previously been reported by Altilia et al. [19] as a cause of defective function of ABCA1, while Y1532C is a novel mutation. Login to comment
69 With respect to the genotypes, the proband`s mother and brother were both heterozygous for mutation R282X, while his father and daughter were heterozygous for mutation Y1532C. Login to comment
84 Together with the finding that none of 100 healthy unrelated individuals with levels of total serum cholesterol ≤6.5 mmol/l, HDL cholesterol between 1.2 and 1.8 mmol/l and triglycerides ≤3.0 mmol/l were found to be heterozygous for R282X or Y1532C in the ABCA1 gene, our findings suggest that the proband had Tangier disease. Login to comment
89 Mutation R282X has recently been described in a Tangier disease patient, who was also heterozygous for mutation IVS2+5G>C [19]. Login to comment
90 R282X introduces a premature stop codon in the predicted large 1st extracellular loop of ABCA1. Login to comment
91 mRNA analyses of fibroblasts failed to detect a transcript from the allele harboring mutation R282X, which the authors attributed to nonsense-mediated mRNA Fig. 3. Login to comment
98 Thus, R282X should be considered to be a pathogenic mutation. Login to comment
100 It cannot be excluded that this mutation could be a normal genetic variant and that Tangier disease in the proband was caused by mutation R282X and an unidentified mutation in the ABCA1 gene. Login to comment
124 Albrecht et al. [23] showed that mutation L1379F, which is also predicted to be in the 4th extracellular loop, was present at reduced levels at the cell surface. Login to comment
126 In conclusion, we have identified the first patient with Tangier disease of Norwegian origin, due to compound heterozygosity for the previously described ABCA1 mutation R282X [19], and the novel ABCA1 mutation Y1532C. Login to comment
64 However, DNA sequencing of the ABCA1 gene suggested that the proband was a compound heterozygote for the nonsense mutation R282X (c.844 C>T) in exon 9 and the missense mutation Y1532C (c.4595 A>G) in exon 34. Login to comment
65 R282X has previously been reported by Altilia et al. [19] as a cause of defective function of ABCA1, while Y1532C is a novel mutation. Login to comment
82 Together with the finding that none of 100 healthy unrelated individuals with levels of total serum cholesterol ࣘ6.5 mmol/l, HDL cholesterol between 1.2 and 1.8 mmol/l and triglycerides ࣘ3.0 mmol/l were found to be heterozygous for R282X or Y1532C in the ABCA1 gene, our findings suggest that the proband had Tangier disease. Login to comment
87 Mutation R282X has recently been described in a Tangier disease patient, who was also heterozygous for mutation IVS2+5G>C [19]. Login to comment
88 R282X introduces a premature stop codon in the predicted large 1st extracellular loop of ABCA1. Login to comment
96 Thus, R282X should be considered to be a pathogenic mutation. Login to comment

[hide] Variations on a gene: rare and common variants in ... Annu Rev Nutr. 2006;26:105-29. Brunham LR, Singaraja RR, Hayden MR
Variations on a gene: rare and common variants in ABCA1 and their impact on HDL cholesterol levels and atherosclerosis.
Annu Rev Nutr. 2006;26:105-29., [PMID:16704350]

Abstract [show]
Cholesterol and its metabolites play a variety of essential roles in living systems. Virtually all animal cells require cholesterol, which they acquire through synthesis or uptake, but only the liver can degrade cholesterol. The ABCA1 gene product regulates the rate-controlling step in the removal of cellular cholesterol: the efflux of cellular cholesterol and phospholipids to an apolipoprotein acceptor. Mutations in ABCA1, as seen in Tangier disease, result in accumulation of cellular cholesterol, reduced plasma high-density lipoprotein cholesterol, and increased risk for coronary artery disease. To date, more than 100 coding variants have been identified in ABCA1, and these variants result in a broad spectrum of biochemical and clinical phenotypes. Here we review genetic variation in ABCA1 and its critical role in cholesterol metabolism and atherosclerosis in the general population.

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554 Conversely, a small number of mutations are associated with less than 50% of control HDL cholesterol, specifically M1091T, G1216V, and the truncation mutations R2144X, R282X, and R909X. Login to comment
555 Since a complete loss of function allele would be expected to result in a 50% reduction in HDL levels, a greater than 50% reduction in HDL is most likely explained by a dominant negative allele, in which TABLE 3 Patient phenotypes associated with heterozygous ABCA1 mutations Mutation HDL (mmol/L) HDL (% of control) Number of patients M1091T 0.48 ± 0.5 30 ± 30 4 G1216V 0.50 40 1 R2144X 0.56 ± 0.2 41 ± 18 12 R282X 0.52 41 1 R909X 0.59 ± 0.3 42 ± 19 5 K776N 0.55 ± 0.1 47 ± 5 2 R587W 0.61 ± 0.1 47 ± 8 7 S364C 0.60 48 1 P1065S 0.80 51 1 c-ter deletion 0.75 53 1 N1800H - 56.5 33 P85L 0.72 ± 0.4 57 ± 33 5 Del693L 0.79 ± 0.2 57 ± 15 8 D1289N 0.80 ± 0.1 59 ± 12 4 R2081W 0.80 ± 0.1 59 ± 12 4 2203X 0.80 ± 0.2 59 ± 20 4 DelED1893,4 0.77 ± 0.2 59 ± 18 8 2145X 0.82 ± 0.1 59 ± 9 4 A1046D 0.70 ± 0.1 60 ± 8 2 Q597R 0.82 ± 0.1 60 ± 5 5 C1477R 0.82 ± 0.2 61 ± 15 9 IVS25 + 1G > C 0.78 ± 0.1 62 ± 12 4 D1099Y 0.83 ± 0.3 63 ± 21 5 1552X 1.00 64 1 F2009S 0.82 ± 0.2 64 ± 19 6 R587W 0.86 ± 0.1 65 ± 17 2 R1068H 0.90 ± 0.3 67 ± 26 9 N935S 1.00 ± 0.3 74 ± 16 7 T929I 1.01 ± 0.2 76 ± 7 8 1284X 1.11 ± 0.2 83 ± 14 5 A937V 1.15 ± 0.6 85 ± 28 2 R1680W 1.22 ± 0.2 87 ± 17 3 635X 1.24 ± 0.5 90 ± 32 7 W590S 1.32 ± 0.6 103 ± 46 15 the mutant protein actually interferes with the activity of the remaining wild-type protein. Login to comment

[hide] Genetics of HDL regulation in humans. Curr Opin Lipidol. 2003 Jun;14(3):273-9. Miller M, Rhyne J, Hamlette S, Birnbaum J, Rodriguez A
Genetics of HDL regulation in humans.
Curr Opin Lipidol. 2003 Jun;14(3):273-9., [PMID:12840658]

Abstract [show]
PURPOSE OF REVIEW: To review gene regulation of HDL-cholesterol and discuss molecular abnormalities in HDL candidate genes that may lead to human pathologic states. RECENT FINDINGS: The inverse association between HDL-cholesterol and vascular disease, especially coronary heart disease, has long been recognized, but understanding gene regulation of HDL in humans gained considerable momentum following the identification of ABCA1 as playing a pivotal role in reverse cholesterol transport. Recent data suggest that potentially important targets for upregulating HDL in humans include upregulators of ABCA1 and APOA1 (e.g. peroxisome proliferator activated receptor and liver X receptor agonists) and downregulators of CETP (e.g. JTT-705). A host of other nuclear receptors under investigation in animal models may advance to human testing in the near future. SUMMARY: Disorders affecting HDL metabolism are complex because monogenic disorders causing low HDL do not necessarily correlate with premature vascular disease. To date, pathologic phenotypes have only been deduced among several HDL candidate genes. Understanding the genetic underpinnings associated with variant HDL and reverse cholesterol transport provides an exceptional opportunity to identify novel agents that may optimize this process and reduce vascular event rates beyond currently available LDL lowering therapies.

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65 TD 1210-1217 8bp del 9 277X truncation [65] TD 1239 C/T 9 R282X truncation [73.] Login to comment

[hide] Abnormal splicing of ABCA1 pre-mRNA in Tangier dis... J Lipid Res. 2003 Feb;44(2):254-64. Epub 2002 Nov 4. Altilia S, Pisciotta L, Garuti R, Tarugi P, Cantafora A, Calabresi L, Tagliabue J, Maccari S, Bernini F, Zanotti I, Vergani C, Bertolini S, Calandra S
Abnormal splicing of ABCA1 pre-mRNA in Tangier disease due to a IVS2 +5G>C mutation in ABCA1 gene.
J Lipid Res. 2003 Feb;44(2):254-64. Epub 2002 Nov 4., [PMID:12576507]

Abstract [show]
Two point mutations of ABCA1 gene were found in a patient with Tangier disease (TD): i) G>C in intron 2 (IVS2 +5G>C) and ii) c.844 C>T in exon 9 (R282X). The IVS2 +5G>C mutation was also found in the brother of another deceased TD patient, but not in 78 controls and 33 subjects with low HDL. The IVS2 +5G>C mutation disrupts ABCA1 pre-mRNA splicing in fibroblasts, leading to three abnormal mRNAs: devoid of exon 2 (Ex2-/mRNA), exon 4 (Ex4-/mRNA), or both these exons (Ex2-/Ex4-/mRNA), each containing a translation initiation site. These mRNAs are expected either not to be translated or generate short peptides. To investigate the in vitro effect of IVS2 +5G>C mutation, we constructed two ABCA1 minigenes encompassing Ex1-Ex3 region, one with wild-type (WTgene) and the other with mutant (MTgene) intron 2. These minigenes were transfected into COS1 and NIH3T3, two cell lines with a different ABCA1 gene expression. In COS1 cells, WTgene pre-mRNA was spliced correctly, while the splicing of MTgene pre-mRNA resulted in Ex2-/mRNA. In NIH3T3, no splicing of MTgene pre-mRNA was observed, whereas WTgene pre-mRNA was spliced correctly. These results stress the complexity of ABCA1 pre-mRNA splicing in the presence of splice site mutations.

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1 254 Journal of Lipid Research Volume 44, 2003 This article is available online at http://www.jlr.org Abnormal splicing of ABCA1 pre-mRNA in Tangier disease due to a IVS2 ϩ5GϾC mutation in ABCA1 gene Serena Altilia,* Livia Pisciotta,† Rita Garuti,* Patrizia Tarugi,* Alfredo Cantafora,§ Laura Calabresi,** Jacopo Tagliabue,†† Sergio Maccari,§§ Franco Bernini,*** Ilaria Zanotti,*** Carlo Vergani,†† Stefano Bertolini,† and Sebastiano Calandra1,** Department of Biomedical Sciences,* University of Modena and Reggio Emilia; Department of Internal Medicine,† University of Genoa; National Institute of Health,§ Rome; Department of Pharmacological Sciences,** Center "E. Grossi Paoletti," University of Milan; Ospedale Maggiore-IRCCS,†† Milan; Sant`Anna Hospital,§§ Castelnuovo Monti, Reggio Emilia; and Departments of Pharmacological*** and Biological Sciences and Applied Chemistries, University of Parma, Italy Abstract Two point mutations of ABCA1 gene were found in a patient with Tangier disease (TD): i) GϾC in intron 2 (IVS2 ؉5GϾC) and ii) c.844 CϾT in exon 9 (R282X). Login to comment
79 The R282X mutation and the cosegregating haplotype are indicated by red color (see sequence analysis in Fig. 3). Login to comment
165 The sequence of ABCA1 gene revealed that proband I.1 (Family 1) was a compound heterozygote for: i) a GϾC transversion at the 5th nucleotide of intron 2 (IVS2 ϩ5GϾC), and ii) a c.844 CϾT transition in exon 9, which converts the arginine codon at position 282 into a termination codon (R282X) (Fig. 3). Login to comment
168 The c.844 CϾT mutation (R282X) introduces a new AflIII restriction site. Login to comment
170 The proband`s brother was heterozygous for the nonsense mutation in exon 9 (R282X). Login to comment
174 Haplotype analysis The genotyping of five polymorphic markers close to or within the ABCA1 gene demonstrated: i) the IVS2 ϩ5GϾC mutation co-segregates with haplotype a; ii) the probands of Family 1 and Family 2 share this haplotype; iii) the nonsense mutation (R282X) cosegregates with haplotype b (Fig. 1). Login to comment
179 Skin fibroblasts from a control subject, proband I.1 of Family 1 (IVS2 ϩ5GϾC/ R282X), and another TD patient (homozygous for R587W) reported previously (13) were labeled with [3H]cholesterol and then incubated in the presence or in the absence of 22-hydroxycholesterol and 9-cis- retinoic acid (22ch/RA). Login to comment
185 Analysis of cDNA Unexpectedly, the sequence of the cDNA region encompassing exons 5-10 failed to confirm the presence of the c.844 CϾT (R282X) mutation in exon 9 in proband I.1 (Family 1), suggesting that only one mutant allele was expressed in fibroblasts. Login to comment
211 ABCA1 cDNA region spanning from exon 5 to exon 10 was amplified by PCR and digested with Af l III to detect the c.844 CϾT mutation (R282X) in exon 9. Login to comment
274 One of the mutations found in proband I.1 (Family 1) is a nonsense mutation in exon 9 (R282X) that involves a CpG dinucleotide, a well-recognized hot spot for base substitution (39). Login to comment
0 Copyright (c) 2003 by Lipid Research, Inc. 254 Journal of Lipid Research Volume 44, 2003 This article is available online at http://www.jlr.org Abnormal splicing of ABCA1 pre-mRNA in Tangier disease due to a IVS2 5GC mutation in ABCA1 gene Serena Altilia,* Livia Pisciotta,ߤ Rita Garuti,* Patrizia Tarugi,* Alfredo Cantafora,&#a7; Laura Calabresi,** Jacopo Tagliabue,ߤߤ Sergio Maccari,&#a7;&#a7; Franco Bernini,*** Ilaria Zanotti,*** Carlo Vergani,ߤߤ Stefano Bertolini,ߤ and Sebastiano Calandra1,** Department of Biomedical Sciences,* University of Modena and Reggio Emilia; Department of Internal Medicine,ߤ University of Genoa; National Institute of Health,&#a7; Rome; Department of Pharmacological Sciences,** Center "E. Grossi Paoletti," University of Milan; Ospedale Maggiore-IRCCS,ߤߤ Milan; Sant`Anna Hospital,&#a7;&#a7; Castelnuovo Monti, Reggio Emilia; and Departments of Pharmacological*** and Biological Sciences and Applied Chemistries, University of Parma, Italy Abstract Two point mutations of ABCA1 gene were found in a patient with Tangier disease (TD): i) GC in intron 2 (IVS2 5GC) and ii) c.844 CT in exon 9 (R282X). Login to comment
78 The R282X mutation and the cosegregating haplotype are indicated by red color (see sequence analysis in Fig. 3). Login to comment
164 The sequence of ABCA1 gene revealed that proband I.1 (Family 1) was a compound heterozygote for: i) a GC transversion at the 5th nucleotide of intron 2 (IVS2 5GC), and ii) a c.844 CT transition in exon 9, which converts the arginine codon at position 282 into a termination codon (R282X) (Fig. 3). Login to comment
167 The c.844 CT mutation (R282X) introduces a new AflIII restriction site. Login to comment
169 The proband`s brother was heterozygous for the nonsense mutation in exon 9 (R282X). Login to comment
173 Haplotype analysis The genotyping of five polymorphic markers close to or within the ABCA1 gene demonstrated: i) the IVS2 5GC mutation co-segregates with haplotype a; ii) the probands of Family 1 and Family 2 share this haplotype; iii) the nonsense mutation (R282X) cosegregates with haplotype b (Fig. 1). Login to comment
178 Skin fibroblasts from a control subject, proband I.1 of Family 1 (IVS2 5GC/ R282X), and another TD patient (homozygous for R587W) reported previously (13) were labeled with [3H]cholesterol and then incubated in the presence or in the absence of 22-hydroxycholesterol and 9-cis-retinoic acid (22ch/RA). Login to comment
184 Analysis of cDNA Unexpectedly, the sequence of the cDNA region encompassing exons 5-10 failed to confirm the presence of the c.844 CT (R282X) mutation in exon 9 in proband I.1 (Family 1), suggesting that only one mutant allele was expressed in fibroblasts. Login to comment
210 ABCA1 cDNA region spanning from exon 5 to exon 10 was amplified by PCR and digested with Af l III to detect the c.844 CT mutation (R282X) in exon 9. Login to comment
273 One of the mutations found in proband I.1 (Family 1) is a nonsense mutation in exon 9 (R282X) that involves a CpG dinucleotide, a well-recognized hot spot for base substitution (39). Login to comment