ABCMdb

ABCC1 p.Gly756Asp

None has been submitted yet.

Publications

PMID: 11466279 [PubMed] Falcon-Perez JM et al: "Domain interactions in the yeast ATP binding cassette transporter Ycf1p: intragenic suppressor analysis of mutations in the nucleotide binding domains."

No. Sentence Comment

55 To determine whether the reversion occurred at the site of the original mutation, the DNA regions that include the primary mutation were sequenced in the plasmids rescued from revertants of G663V, G756D, and G1306E mutations. Login to comment
101 For the intragenic suppressor analysis of mutations localized in the NBDs, we selected G663V, G756D, D777N, G1306E, and G1311R changes (Fig. 1), all of which affect residues in highly conserved motifs. Login to comment
108 Mutagenesis of plasmids containing the G663V, G756D, D777N, and G1306E mutations resulted in transformants that grew in the presence of Cd2ϩ , whereas mutagenesis of a plasmid containing the G1311R allele did not (Table 1). Login to comment
111 All revertants of G663V, G756D, and G1306E mutants were full revertants of the initial mutation, and 30 out of 83 revertants isolated for the D777N mutant were due to a second-site suppressor mutation. Login to comment
130 of revertants Full revertants Second-site suppressor mutants G663V Episomal Mutator strain 16 ϫ 104 18 0 G756D Centromeric Chemical 5 ϫ 104 0 0 Episomal Mutator strain 5 ϫ 104 2 0 Chemical 3 ϫ 104 15 0 D777N Episomal Mutator strain 16 ϫ 104 53 30 G1306E Centromeric Chemical 3 ϫ 104 1 0 Mutator strain 5 ϫ 104 4 0 G1311R Centromeric Chemical 3 ϫ 104 0 0 Episomal Chemical 3 ϫ 104 0 0 TABLE 2. Login to comment
204 We performed a revertant analysis of five mutations located in the NBDs of Ycf1p, namely G663V, G756D, D777N, G1306E, and G1311R. Login to comment
207 We were unable to identify any intragenic suppressors of the other four alleles, G663V, G756D, G1306E, and G1311R. Login to comment
212 Moreover, the NBD1 mutations G663V and G756D were not suppressed when combined with some of the suppressors isolated for D777N (see below). Login to comment
230 The specificity of the suppression is further supported by the fact that neither the W1225C nor R1415G mutations were able to suppress the defective growth or Ycf1p transport function of the other NBD1 primary mutations, namely G663V and G756D (data not shown). Login to comment

PMID: 9379898 [PubMed] Wemmie JA et al: "Mutational analysis of the Saccharomyces cerevisiae ATP-binding cassette transporter protein Ycf1p."

No. Sentence Comment

133 These mutants corresponded to CFTR alterations known to be associated with cystic fibrosis (G551D and G551S in CFTR, G756D and G756S in Ycf1p) as well as lesions that either disturb normal function (K464M in CFTR, K669M in Ycf1p) or act to suppress the phenotype of ⌬F508 CFTR (R553Q and R553M in CFTR, K758Q and K758M in CFTR). Login to comment
135 K669M, G756S and G756D were unable to complement the cadmium hypersensitivity of the ⌬ycf1 strain. Western blot analysis of these mutant Ycf1p derivatives indicated that all these proteins were produced at the same level in the cell. Login to comment
237 The three mutant Ycf1p derivatives (⌬F713, G756D and G756S) that correspond to known CF-causing alleles of CFTR (⌬F508, G551D and G551S) all produce a Ycf1p mutant that exhibits a defect analogous to its CFTR counterpart. Login to comment
281 The 2 ␮m plasmids carrying the indicated mutant forms of YCF1 and the relevant mutagenic PCR primers are listed below: pJAW100 (G756S), 5Ј-ATC TCC TTA TCT GGA TCC CAA AAA GCT CGT TTG-3Ј; pJAW88 (G756D), 5Ј-ATC TCC TTA TCT GGA GAC CAA AAA GCT CGT TTG-3Ј; pJAW86 (K669M), 5Ј- AAA GTT GGC AGT GGT ATG ACA GCT CTA TT-3Ј; pJAW98 (K758R), 5Ј-TTA TCT GGA GGA CAA CGG GCC CGT TTG TCT TTA-3Ј; pJAW108 (K758M), 5Ј-AGA CAA ACG AGC CAT TTG TCC TCC AGA TAA GGA TAT CCC TTT C-3Ј; pJAW109 (K758Q), 5Ј-AGA CAA ACG AGC TTG TTG TCC TCC AGA TAA GCT TAT CCC TTT C-3Ј. Login to comment

PMID: 26606940 [PubMed] Wei S et al: "Long-range coupling between the extracellular gates and the intracellular ATP binding domains of multidrug resistance protein pumps and cystic fibrosis transmembrane conductance regulator channels."

No. Sentence Comment

70 Primer sequences for cloning and site-directed mutagenesis Ycf1p Forward cloning primer: CAACACAGGCATGTATATTA- AGAGC Reverse cloning primer: TTAAACTTATGGCGTCAGAG- TTGCC F565A: CATTGACTACTGACTTAGTTGCCCCTGCTTTG- ACTCTGTTC F565S: CATTGACTACTGACTTAGTTTCCCCTGCTTTGA- CTCTGTTC F565L: CATTGACTACTGACTTAGTTTTACCTGCTTTG- ACTCTGTTC G756D: AAGACAAACGAGCTTTTTGATCTCCAGATAAG- GAGATCCC D777N: ACAGCTGGCAAAGGATCATTAAGTAAATAAG- TGTCAGCTC Y1281G: GATCAAGCTCCGGCCTACCACGAGTGGAATA- ATTATTAAAC Yor1p Forward cloning primer: CTAATTGTACATCCGGTTTT- AACC Reverse cloning primer: TTGAGTCATTGCCCTTAA- AATGG F468S: AGGCAACCTGGTAATATTTCTGCCTCTTTATC- TTTATTTC F468A: AGGCAACCTGGTAATATTGCTGCCTCTTTATC- TTTATTTC F468L: AGGCAACCTGGTAATATTCTTGCCTCTTTATC- TTTATTTC G713D: GTGGTATTACTTTATCTGGTGATCAAAAGGCA- CGTATCAATTT Y1222G: ATAGGTAAACCAGGTCTACCGGCAAAATCAA- CATTTTCAA CFTR Forward cloning primer: GAAGAAGCAATGGAAAAA- ATGATTG Reverse cloning primer: TCGGTGAATGTTCTGACCT- TGG F337S: TCATCCTCCGGAAAATATCCACCACCATCTCA- TTCTGC F337A: TCATCCTCCGGAAAATAGCCACCACCATCTCA- TTCTGC F337L: TCATCCTCCGGAAAATATTAACCACCATCTCA- TTCTGC F337C: TCATCCTCCGGAAAATATGCACCACCATCTC- ATTCTGC Immunoblot analysis of CFTR protein expression Expression of the CFTR F337 mutants was verified by immunoblotting as described elsewhere (15). Login to comment
88 For reference, we also introduced each phenylalanine mutation into the WT Ycf1p background and into an NBD1 signature sequence mutant that is expected to have negligible ATPase activity (G756D in Ycf1p) (16, 37, 38). Login to comment
108 Mutating the conserved extracellular F565 residue partially restores the function of ATP binding mutants of Ycf1p (Y1281G and D777N), but not of a signature sequence mutant (G756D). Login to comment
110 The WT and ATP binding mutations, Y1281G (top panel) and D777N (middle), or the signature sequence mutation, G756D (bottom), are indicated along with the F565 substitutions, introduced alone or in combination. Login to comment
111 The vertical rectangular box indicates spot cultures analyzed in B and C. B) Restoration of Ycf1p function is observed as differential growth associated with F565 substitutions in the context of the Y1281G (top) and D777N (middle) loss of function mutations but not the G756D signature sequence mutation (bottom). Login to comment
114 C) Functional comparison between each single mutant, Ycf1p-Y1281G (top), Ycf1p-D777N (middle) or Ycf1p-G756D (bottom), and the respective A, S, and L substitutions for F565. Login to comment
69 Primer sequences for cloning and site-directed mutagenesis Ycf1p Forward cloning primer: CAACACAGGCATGTATATTA- AGAGC Reverse cloning primer: TTAAACTTATGGCGTCAGAG- TTGCC F565A: CATTGACTACTGACTTAGTTGCCCCTGCTTTG- ACTCTGTTC F565S: CATTGACTACTGACTTAGTTTCCCCTGCTTTGA- CTCTGTTC F565L: CATTGACTACTGACTTAGTTTTACCTGCTTTG- ACTCTGTTC G756D: AAGACAAACGAGCTTTTTGATCTCCAGATAAG- GAGATCCC D777N: ACAGCTGGCAAAGGATCATTAAGTAAATAAG- TGTCAGCTC Y1281G: GATCAAGCTCCGGCCTACCACGAGTGGAATA- ATTATTAAAC Yor1p Forward cloning primer: CTAATTGTACATCCGGTTTT- AACC Reverse cloning primer: TTGAGTCATTGCCCTTAA- AATGG F468S: AGGCAACCTGGTAATATTTCTGCCTCTTTATC- TTTATTTC F468A: AGGCAACCTGGTAATATTGCTGCCTCTTTATC- TTTATTTC F468L: AGGCAACCTGGTAATATTCTTGCCTCTTTATC- TTTATTTC G713D: GTGGTATTACTTTATCTGGTGATCAAAAGGCA- CGTATCAATTT Y1222G: ATAGGTAAACCAGGTCTACCGGCAAAATCAA- CATTTTCAA CFTR Forward cloning primer: GAAGAAGCAATGGAAAAA- ATGATTG Reverse cloning primer: TCGGTGAATGTTCTGACCT- TGG F337S: TCATCCTCCGGAAAATATCCACCACCATCTCA- TTCTGC F337A: TCATCCTCCGGAAAATAGCCACCACCATCTCA- TTCTGC F337L: TCATCCTCCGGAAAATATTAACCACCATCTCA- TTCTGC F337C: TCATCCTCCGGAAAATATGCACCACCATCTC- ATTCTGC Immunoblot analysis of CFTR protein expression Expression of the CFTR F337 mutants was verified by immunoblotting as described elsewhere (15). Login to comment
87 For reference, we also introduced each phenylalanine mutation into the WT Ycf1p background and into an NBD1 signature sequence mutant that is expected to have negligible ATPase activity (G756D in Ycf1p) (16, 37, 38). Login to comment
107 Mutating the conserved extracellular F565 residue partially restores the function of ATP binding mutants of Ycf1p (Y1281G and D777N), but not of a signature sequence mutant (G756D). Login to comment
109 The WT and ATP binding mutations, Y1281G (top panel) and D777N (middle), or the signature sequence mutation, G756D (bottom), are indicated along with the F565 substitutions, introduced alone or in combination. Login to comment
113 C) Functional comparison between each single mutant, Ycf1p-Y1281G (top), Ycf1p-D777N (middle) or Ycf1p-G756D (bottom), and the respective A, S, and L substitutions for F565. Login to comment