ABCMdb

PMID: 10762539

Mickle JE, Milewski MI, Macek M Jr, Cutting GR
Effects of cystic fibrosis and congenital bilateral absence of the vas deferens-associated mutations on cystic fibrosis transmembrane conductance regulator-mediated regulation of separate channels.
Am J Hum Genet. 2000 May;66(5):1485-95. Epub 2000 Apr 4., [PubMed]

Sentences

No. Mutations Sentence Comment

37 ABCC7 p.Gly551Asp ABCC7 p.Ala455Glu Specifically, two missense mutations in NBF1 have been evaluated: A455E, which is associated with mild lung disease, and G551D, which is associated with a more severe pulmonary phenotype (Hamosh et al. 1992; Gan et al. 1995). Login to comment 38 ABCC7 p.Ala455Glu CFTR bearing A455E retains both CFTR Cl2 channel activity and the ability to regulate ORCCs (Fulmer et al. 1995). Login to comment 39 ABCC7 p.Gly551Asp In contrast, G551D-CFTR has some Cl2 channel activity but does not regulate ORCCs (Fulmer et al. 1995). Login to comment 40 ABCC7 p.Gly551Asp It has been demonstrated that G551D-CFTR does not regulate the activity of ENaC (Ismailov et al. 1996). Login to comment 47 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Arg334Trp ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr DNA was assayed for 16 common CFTR mutations (R117H, 62111GrT, R334W, R349P, A455E, DI507, DF508, 1717-1GrA, G542X, S549N, G551D, R553X, R560T, 3849110 Kb CrT, W1282X, and N1303K), by reverse dot-blot hybridization (Mickle et al. 1998). Login to comment 50 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn ABCC7 p.Arg1070Trp Expression Analysis The mutations DF508, R1070W, D1270N, and G1349D were created in the vector pBQ4.7 containing CFTR cDNA (pBQ4.7 is a gift from J. Rommens and L. Login to comment 51 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn ABCC7 p.Arg1070Trp C. Tsui), by single-stranded mutagenesis (Youssoufian et al. 1995), and then were shuttled into pRSV-CFTR, a Rous sarcoma virus (RSV)-driven expression plasmid, by use of Kpn2I and HpaI (for DF508) or NcoI and SalI (for R1070W, D1270N, and G1349D) restriction sites common to both plasmids (Fulmer et al. 1995). Login to comment 52 ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Pro The mutations R1070P and R1070Q were created directly in pRSV-CFTR, by use of a transformer site-directed mutagenesis kit (Clontech). Login to comment 55 ABCC7 p.Trp1282* IB3-1 bronchial epithelial cells were derived from a patient with CF (genotype DF508/W1282X); these cells lack functional CFTR (Zeitlin et al. 1991). Login to comment 68 ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Pro Mutations at codon 1070 of TMD2 were selected, since two mutations (R1070P and R1070Q) have been associated with CF, whereas a third (R1070W) has been observed in men with CBAVD (table 1). Login to comment 69 ABCC7 p.Arg1070Trp The R1070W mutation was first reported by us to the Cystic Fibrosis Genetic Analysis Consortium. Login to comment 74 ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Trp The latter mutation is predicted to change the amino acid at residue 1070 from arginine to tryptophan and is designated "R1070W." Login to comment 75 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn Also selected for study were two mutations in NBF2-D1270N and G1349D, which are associated with different pulmonary phenotypes (table 1). Login to comment 76 ABCC7 p.Asp1270Asn The mutation D1270N has been identified in at least nine men with CBAVD. Login to comment 77 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn For comparison purposes, we selected the CF-associated mutation G1349D, since it occurs in the same functional domain as does D1270N. Login to comment 91 ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Trp Thus, mutations R1070Q and R1070W altered but did not prohibit complex glycosylation. Login to comment 97 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Pro OF CASES PHENOTYPE Lung Status Pancreatic Status Sweat Cl2 Fertility Normala Abnormal Not Reported Sufficient Insufficient Not Reported Reported (Mean 5 SEM [mmol/literb ]) Not Reported Subfertilec Not Reported R1070W (7)d 5 0 2 5 0 2e 6 ( ) 50.2 5 13.4 1 6 1 CBAVD R1070P (2)f 0 1 1 0 1 1 1 (Positive) 1 0 2 CF R1070Q (14)g 0 7 7 0 7 7 7 (Positive) 7 2 12 CF D1270N (9)h 4 0 5 4 0 5 3 ( ) 77.5 5 16.7 6 3 6 CBAVD G1349D (3)i 0 0 3 0 0 3 ) 3 1 2 CF a No history of chronic lung disease. Login to comment 134 ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp The initial mutation report (Beaudet et al. 1991) indentified G1349D on two CF chromosomes, and, on the basis of these cases, G1349D has been described as a CF-associated mutation (Gregory et al. 1991; Anderson and Welsh 1992), with specific reference to PI (Welsh and Smith 1993). Login to comment 172 ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Pro B, CBAVD(R1070W)- and CF(R1070P and R1070Q)-associated mutants in TMD2 had I-V plots similar to those of wild-type CFTR: outwardly rectified currents (blackened circles) that responded to DIDS (unblackened circles) and glibenclamide (crosses). Login to comment 173 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn C, CBAVD(D1270N) and CF(G1349D) mutations in NBF2 had I-V profiles comparable to those of wild-type CFTR. Login to comment 180 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Pro Thus, the combination of I-V relationship and response to inhibitors allowed dissection of whole-cell Cl-currents into two components: outwardly rectified and DIDS sensitive, carried by separate channels such as ORCCs; and linear, DIDS insensitive, and gli- Table 2 Summary of Processing and Whole-Cell Function of CFTR Mutants DOMAIN AND MUTATION PHENOTYPE CFTR STATUS a Processingb Function Band A Band B Band C Cl2 Channel Regulatoryc Not applicable: Wild type Normal 2 2 111 111 1 NBF1:d A455Ee CFe 1 11 2 111 1 DF508 CF 1 11 2 1 2 G551D CF 2 2 111 1 2 TMD2: R1070W CBAVD 2 1 11 111 1 R1070P CF 1 11 2 111 1 R1070Q CF 2 1 11 111 1 NBF2: D1270N CBAVD 2 2 111 111 1 G1349D CF 2 2 111 111 1 a A minus sign (2) denotes absence; a single plus sign (1) denotes "low"; a double plus sign (11) denotes "intermediate"; and a triple plus sign denotes "high." Login to comment 182 ABCC7 p.Gly551Asp ABCC7 p.Ala455Glu d Data for A455E and G551D have been reported elsewhere (Fulmer et al. 1995). Login to comment 188 ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Pro .05 mutant (R1070W) and the CF mutants (R1070P and R1070Q) in TMD2 generated outwardly rectified Cl2 currents that were inhibited by DIDS (fig. 3B). Login to comment 190 ABCC7 p.Gly1349Asp ABCC7 p.Asp1270Asn Likewise, cells expressing either of the NBF2 mutants- D1270N (CBAVD) and G1349D (CF)-had both components of whole-cell Cl2 currents (fig. 3C). Login to comment 196 ABCC7 p.Arg1070Pro We were unable to detect mature glycosylated forms for two mutants, DF508 and R1070P; yet, each of these generated Cl2 currents attributed to CFTR activity. Login to comment 213 ABCC7 p.Gly551Asp For example, CF-associated mutations in the NBF1 domain, DF508 (present study), and G551D (Fulmer et al. 1995), ablated CFTR regulation of ORCCs, whereas the regulatory ability was not impaired by mutations in TMD2 and NBF2 (table 2). Login to comment 214 ABCC7 p.Gly1349Asp Of particular note are the results obtained with the NBF2 mutation G1349D. Login to comment 215 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp The latter mutation is comparable, in terms of nature (glycine to aspartic acid) and location (Walker C motif), to the NBF1 mutation G551D; yet, CFTR bearing G1349D-generated DIDS-sensitive currents that were attributed to the regulation of ORCCs. Login to comment 230 ABCC7 p.Arg1070Trp Electronic-Database Information Accession numbers and URLs for data in this article are as follows: Cystic Fibrosis Genetic Analysis Consortium, http://www .genet.sickkids.on.ca/CFTR (for R1070W) Online Mendelian Inheritance in Man (OMIM), http://www .ncbi.nim.nih.gov/Omim (for CF [MIM 219700], CFTR [MIM 602421], and CBAVD [MIM 277180]) References Anderson MP, Rich DP, Gregory RJ, Smith AE, Welsh MJ (1991) Generation of cAMP-activated chloride currents by expression of CFTR. Login to comment