ABCMdb

ABCC7 p.Lys1218Arg

None has been submitted yet.

Publications

PMID: 24777605 [PubMed] Lee S et al: "Interference with ubiquitination in CFTR modifies stability of core glycosylated and cell surface pools."

No. Sentence Comment

7 The K1218R mutant increases total and cell surface CFTR, which is further accumulated by proteasomal and lysosomal inhibition. Login to comment
73 We hypothe- TABLE 1 Primers for site-directed mutagenesis Mutation Directiona Sequence (5=-3=)b K14R F GGCCAGCGTTGTCTCCAGACTTTTTTTCAGCTGGACC R GGTCCAGCTGAAAAAAAGTCTGGAGACAACGCTGGCC K68R F GGCTTCAAAGAAAAATCCTAGACTCATTAATGCCCTTCGGCG R CGCCGAAGGGCATTAATGAGTCTAGGATTTTTCTTTGAAGCC K710R F CCAATCAACTCTATACGAAGATTTTCCATTGTGCAAAAG R CTTTTGCACAATGGAAAATCTTCGTATAGAGTTGATTGG K716R F GAAAATTTTCCATTGTGCAAAGGACTCCCTTACAAATGAATGG R CCATTCATTTGTAAGGGAGTCCTTTGCACAATGGAAAATTTTC K710/716R F CCAATCAACTCTATACGAAGATTTTCCATTGTGCAAAGGACTCCCTTACAAATGAATGG R CCATTCATTTGTAAGGGAGTCCTTTGCACAATGGAAAATCTTCGTATAGAGTTGATTGG K1041R F CCTCACAGCAATTCAGACAACTGGAATCTGAAG R CTTCAGATTCCAGTTGTCTGAGTTGCTGTGAGG K1080R F GAAACTCTGTTCCACAGAGCTCTGAATTTACATAC R GTATGTAAATTCAGAGCTCTGTGGAACAGAGTTTC K1218R F GATCTCACAGCAAGATACACAGAAGG R CCTTCTGTGTATCTTGCTGTGAGATC a F, forward; R, reverse. Login to comment
102 The K1218R mutation increases CFTR B and C bands even more significantly than N-tail mutants (Fig. 2B). Login to comment
130 This immunoblot demonstrates increased levels of band C CFTR expression from the K14R, K68R, and K1218R cDNA vectors and decreased levels of band C CFTR expression in K710R, K716R, K710/716R, and K1080R vectors. Login to comment
157 K1080R and K1218R mutants displayed both bands B and C after proteasomal inhibition. Login to comment
158 This supports the hypothesis that the K1080R and K1218R mutants undergo the same degree of proteasomal degradation as wild-type CFTR and are still trafficked. Login to comment
169 (C) The results from expression of control vector, wt CFTR, K1041R, K1080R, and K1218R with and without proteasomal or lysosomal inhibition are shown in representative blots similar to the results show in panel A. MG132 rescues band B from K1041R to some extent, as well as a small portion of band C. Login to comment
171 K1218R shows the most robust rescue by both. Login to comment
174 pression of K1080R C band and improved the K1218R mutant, indicating that the K1218R mutant can avoid lysosomal degradation. Login to comment
191 The K710R, K716R, and K710/716R mutants showed faster degradation of the C band of CFTR than the wild type and K1218R. Login to comment
193 The initial expression level of K1218R is higher than that of wt CFTR, and the degradation rate of K1218R is similar to that of the wt CFTR. Login to comment
194 K14R, K1080R, and K1218R but not K1041R reach the cell surface. Login to comment
200 There were decreased levels of cell surface CFTR detection in the K68R, K710R, K716R, K710/ K716R, and K1041R mutant forms of CFTR, but the wild-type, K14R, K1080R, and K1218R mutants accumulated similar levels of visible cell surface CFTR. Login to comment
201 Analysis of the Z stack of images for the wild type, K68R, K710R, K1080R, and K1218R confirmed that the CFTR immunostaining, shown in red, resided at the apical surface in these nonpermeabilized cells (data not shown). Login to comment
208 As expected, increased amounts of K1218R were detected from the cell surface fraction (Fig. Login to comment
214 Preventing ubiquitination at K14R, K1080R, and K1218R allows the mutant protein to gain access to the cell surface. Login to comment
218 Control vector, wt CFTR, K14R, K68R, K710R, K716R, K710/716R, K1041R, K1080R, and K1218R were expressed in IB3-1 cells for 48 h. CFTR was immunoprecipitated (IP) with M3A7 as described in Materials and Methods and separated by SDS-PAGE. Login to comment
221 K63-linked ubiquitin on CFTR is reduced in the K68R, K710R, K716R, and K1218R mutants. Login to comment
226 FIG 6 K-to-R mutation alters the stability of CFTR protein, and wt CFTR, K14R, K1080R, and K1218R, but not K1041R, are detectable at the cell surface. Login to comment
230 Wild-type and K1218R CFTR maintain the highest stability with little turnover in 24 h. Login to comment
237 CFTR is easily detected HDAC6 must interact with CFTR to promote trafficking beyond the ER, then we predict that K1041R would interact preferentially with VCP and that K1218R would interact preferentially with HDAC6. Login to comment
241 HDAC6 was abundant in all cases, and ZO-1 was conspicuously absent from the K1080R and K1218R mutants. Login to comment
247 K1218R is the most interesting construct since this mutant CFTR does better than the wild type in remaining at the cell surface in large-mass quantities. Login to comment
252 The F508del from the surface of cells expressing wt CFTR, K14R, K1080R, and K1218R. Login to comment
261 Band C was most reduced in K1041R and highest in the wt, K1080R, and K1218R, consistent with data in panels A and B. Login to comment
267 Cell surface band C, prominent for wt CFTR and K1218R located on the PM, was unaffected by MG132 and increased by lysosomal inhibition. Login to comment
269 A similar pattern was seen with lysosomal inhibition of cells expressing K1218R, suggesting that there is a ubiquitination step between the cell surface and the lysosome occurring through an alternate site. Login to comment
274 We did not detect ZO-1 in the K1080R or K1218R complex. Login to comment
275 Since ZO-1 interacts with PDZ domains, the reduced interactions in K14R, K68R, K710R, K716R, K710/716R, and K1041R and the absence of interactions with K1080R and K1218R raise the issue of alternative trafficking routes that are not dependent on the C-tail PDZ domain. Login to comment
280 K1218R bypasses the proteasome fairly efficiently and further is slow to undergo lysosomal degradation. Login to comment
296 Interestingly, ZO-1 was highly abundant in complexes formed with wt CFTR; K14R, K68R, K710R, K716R, K710/716R, and K1041R mutants bound less ZO-1, whereas K1080R and K1218R mutant complexes had none. Login to comment