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Sorting Signals, Domain Conformation and Interdomain Interactions in CFTR Misprocessing and Rescue

Bhrigu, Gargi
http://rave.ohiolink.edu/etdc/view?acc_num=mco1268659408

Abstract Details

2010, Doctor of Philosophy in Biomedical Sciences (Ph.D.), University of Toledo, College of Medicine.
Cystic fibrosis (CF) is among the most common lethal inherited diseases in the US. Thedeletion of phenylalanine at residue 508 (6F508) within cystic fibrosis transmembrane conductance regulator (CFTR) is the most common CF causing mutation. 6F508 mutant is unable to exit the endoplasmic reticulum (ER). The precise mechanism of the defective ER-to-Golgi export of 6F508 CFTR is not known. A di-acidic ER exit code (DAD) has been identified within the first nucleotide binding domain (NBD1) of CFTR. Disruption of this motif leads to the defective coupling of CFTR to the COPII machinery and the subsequent impaired export of CFTR from the ER. We performed a systematic analysis of the sorting signals, domain conformation and inter-domain interactions of CFTR in presence of 6F508 mutation and during its rescue by low temperature or second site mutation (R555K). We found that rescue of 6F508 CFTR depends on the DAD motif within the NBD1. Disruption of DAD (DAA) reduces the Sec24 association of 6F508 CFTR whereas rescue mutation R555K increases it. Using in situ limited proteolysis we identified conformational defects in N-terminal region and NBD1 in 6F508 CFTR. 6F508 rescue is accompanied by global conformational reversion. Further domaindomain interactions within CFTR play an important role in both 6F508 CFTR misprocessing and rescue. Our data firmly establish that while DAA is an exit code mutant 6F508 CFTR has widespread conformational defect which compromises its export and stability. Restoration of its global conformation leads to restoration of its export as well as function.
Xiaodong Robert Wang, PhD (Committee Chair)
Andrew Beavis, PhD (Committee Member)
Nikolai Modyanov, PhD (Committee Member)
Ronald Mellgren, PhD (Committee Member)
Bina Joe, PhD (Committee Member)
116 p.
Cellular Biology; Molecular Biology; Physiological Psychology
CFTR; DF508; Protein trafficking; Endoplasmic Reticulum; conformation; exit code

Recommended Citations

Citations

  • Bhrigu, G. (2010). Sorting Signals, Domain Conformation and Interdomain Interactions in CFTR Misprocessing and Rescue [Doctoral dissertation, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=mco1268659408

    APA Style (7th edition)

  • Bhrigu, Gargi. Sorting Signals, Domain Conformation and Interdomain Interactions in CFTR Misprocessing and Rescue. 2010. University of Toledo, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=mco1268659408.

    MLA Style (8th edition)

  • Bhrigu, Gargi. "Sorting Signals, Domain Conformation and Interdomain Interactions in CFTR Misprocessing and Rescue." Doctoral dissertation, University of Toledo, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=mco1268659408

    Chicago Manual of Style (17th edition)

mco1268659408
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© 2010, all rights reserved.
This open access ETD is published by University of Toledo Health Science Campus and OhioLINK.