Mutations in the nucleotide binding domain 1 signature motif region rescue processing and functional defects of cystic fibrosis transmembrane conductance regulator ΔF508

被引:86
作者
deCarvalho, ACV [1 ]
Gansheroff, LJ [1 ]
Teem, JL [1 ]
机构
[1] Florida State Univ, Dept Biol Sci, Biounit 238, Tallahassee, FL 32306 USA
关键词
D O I
10.1074/jbc.M205644200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), an ATP binding cassette (ABC) transporter that functions as a phosphorylationand nucleotide-regulated chloride channel, is mutated in cystic fibrosis (CF) patients. Deletion of a phenylalanine at amino acid position 508 (AF508) in the first nucleotide binding domain (NBD1) is the most prevalent CF-causing mutation and results in defective protein processing and reduced CFTR function, leading to chloride impermeability in CF epithelia and heterologous systems. Using a STE6/CFTRAF508 chimera system in yeast, we isolated two novel AF508 revertant mutations, I539T and G550E, proximal to and within the conserved ABC signature motif of NBD1, respectively. Western blot and functional analysis in mammalian cells indicate that mutations 1539T and G550E each partially rescue the CFTRAF508 defect. Furthermore, a combination of both revertant mutations resulted in a 38-fold increase in CFTRAF508-mediated chloride current, representing 29% of wild type channel activity. The G550E mutation increased the sensitivity of CFTRAF508 and wild type CFTR to activation by cAMP agonists and blocked the enhancement of CFTRAF508 channel activityby.2 mm 3-isobutyl-1-methylxanthine. The data show that the AF508 defect can be significantly rescued by second-site mutations in the nucleotide binding domain 1 region, that includes the LSGGQ consensus motif.
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页码:35896 / 35905
页数:10
相关论文
共 79 条
[1]   Activation of wild type and ΔF508-CFTR by phosphodiesterase inhibitors through cAMP-dependent and -independent mechanisms [J].
Al-Nakkash, L ;
Hwang, TC .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1999, 437 (04) :553-561
[2]   Activation of ΔF508 CFTR in a cystic fibrosis respiratory epithelial cell line by 4-phenylbutyrate, genistein and CPX [J].
Andersson, C ;
Roomans, GM .
EUROPEAN RESPIRATORY JOURNAL, 2000, 15 (05) :937-941
[3]   Contribution of R domain phosphoserines to the function of CFTR studied in Fischer rat thyroid epithelia [J].
Baldursson, O ;
Berger, HA ;
Welsh, MJ .
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2000, 279 (05) :L835-L841
[4]   MUTATIONAL ANALYSIS OF THE YEAST A-FACTOR TRANSPORTER STE6, A MEMBER OF THE ATP BINDING CASSETTE (ABC) PROTEIN SUPERFAMILY [J].
BERKOWER, C ;
MICHAELIS, S .
EMBO JOURNAL, 1991, 10 (12) :3777-3785
[5]   Strategies for correcting the ΔF508 CFTR protein-folding defect [J].
Brown, CR ;
Hong-Brown, LQ ;
Welch, WJ .
JOURNAL OF BIOENERGETICS AND BIOMEMBRANES, 1997, 29 (05) :491-502
[6]   Removal of multiple arginine-framed trafficking signals overcomes misprocessing of ΔF508 CFTR present in most patients with cystic fibrosis [J].
Chang, XB ;
Cui, LY ;
Hou, YX ;
Jensen, TJ ;
Aleksandrov, AA ;
Mengos, A ;
Riordan, JR .
MOLECULAR CELL, 1999, 4 (01) :137-142
[7]   DEFECTIVE INTRACELLULAR-TRANSPORT AND PROCESSING OF CFTR IS THE MOLECULAR-BASIS OF MOST CYSTIC-FIBROSIS [J].
CHENG, SH ;
GREGORY, RJ ;
MARSHALL, J ;
PAUL, S ;
SOUZA, DW ;
WHITE, GA ;
ORIORDAN, CR ;
SMITH, AE .
CELL, 1990, 63 (04) :827-834
[8]   8-cyclopentyl-1,3-dipropylxanthine and other xanthines differentially bind to the wild-type and Delta F508 mutant first nucleotide binding fold (NBF-1) domains of the cystic fibrosis transmembrane conductance regulator [J].
Cohen, BE ;
Lee, G ;
Jacobson, KA ;
Kim, YC ;
Huang, Z ;
Sorscher, EJ ;
Pollard, HB .
BIOCHEMISTRY, 1997, 36 (21) :6455-6461
[9]   GENERATION AND CHARACTERIZATION OF A DELTA-F508 CYSTIC-FIBROSIS MOUSE MODEL [J].
COLLEDGE, WH ;
ABELLA, BS ;
SOUTHERN, KW ;
RATCLIFF, R ;
JIANG, CW ;
CHENG, SH ;
MACVINISH, LJ ;
ANDERSON, JR ;
CUTHBERT, AW ;
EVANS, MJ .
NATURE GENETICS, 1995, 10 (04) :445-452
[10]   CYSTIC-FIBROSIS - MOLECULAR-BIOLOGY AND THERAPEUTIC IMPLICATIONS [J].
COLLINS, FS .
SCIENCE, 1992, 256 (5058) :774-779