A newly independent applicant plans to continue work on mechanisms underlying the defects in rhodopsin mutations. Earlier work by the applicant established two different types of defects in rhodopsin mutants implicated in ADRP. One class is defective in protein exit from the ER/Golgi complex; the second class is incapable of polarized sorting and fails to targeting the rhodopsin to the rod outer segment. In this proposal, the applicant will follow up the observations on this second class of mutants in hopes of deciphering the mechanisms involved in rhodopsin localization. In the first specific aim, three complementary systems will be used to identify the signals responsible for the targeting of rhodopsin to the appropriate membrane. A polarized epithelial cell from dog kidney, the MDCK cell, will be used to analyze large numbers of rhodopsin mutants. To determine the suitability of this system for reflecting photoreceptor behavior, a salamander primary retinal culture will be examined for suitability as an experimental system. Finally, transgenic mice will be used to determine the reliability of both experimental systems in reflecting the in vivo situation. The work will establish the requirement and the sufficiency of the C-terminus domain as well as search for other important sequences in the sorting process. In the second specific aim, other molecular components of the sorting process will be sought by identifying proteins that bind to rhodopsin sorting sequences. Both the yeast two hybrid system and a direct protein/protein filter binding assays will be used. Identified genes will be sequenced to determine the nature of the encoded gene and their subcellular location will be determined.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY011307-02
Application #
2020030
Study Section
Visual Sciences C Study Section (VISC)
Project Start
1996-01-31
Project End
2000-12-31
Budget Start
1997-01-01
Budget End
1997-12-31
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Weill Medical College of Cornell University
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
201373169
City
New York
State
NY
Country
United States
Zip Code
10065
Hsu, Kuo-Shun; Chuang, Jen-Zen; Sung, Ching-Hwa (2017) The Biology of Ciliary Dynamics. Cold Spring Harb Perspect Biol 9:
Saito, Masaki; Otsu, Wataru; Hsu, Kuo-Shun et al. (2017) Tctex-1 controls ciliary resorption by regulating branched actin polymerization and endocytosis. EMBO Rep 18:1460-1472
Mestres, Ivan; Sung, Ching-Hwa (2017) Nervous system development relies on endosomal trafficking. Neurogenesis (Austin) 4:e1316887
Wahl, Silke; Magupalli, Venkat Giri; Dembla, Mayur et al. (2016) The Disease Protein Tulp1 Is Essential for Periactive Zone Endocytosis in Photoreceptor Ribbon Synapses. J Neurosci 36:2473-93
Mestres, Iván; Chuang, Jen-Zen; Calegari, Federico et al. (2016) SARA regulates neuronal migration during neocortical development through L1 trafficking. Development 143:3143-53
Chou, Szu-Yi; Hsu, Kuo-Shun; Otsu, Wataru et al. (2016) CLIC4 regulates apical exocytosis and renal tube luminogenesis through retromer- and actin-mediated endocytic trafficking. Nat Commun 7:10412
Liu, Chenshu; Chuang, Jen-Zen; Sung, Ching-Hwa et al. (2015) A dynein independent role of Tctex-1 at the kinetochore. Cell Cycle 14:1379-88
Hsu, Ya-Chu; Chuang, Jen-Zen; Sung, Ching-Hwa (2015) Light regulates the ciliary protein transport and outer segment disc renewal of mammalian photoreceptors. Dev Cell 32:731-42
Thuenauer, Roland; Hsu, Ya-Chu; Carvajal-Gonzalez, Jose Maria et al. (2014) Four-dimensional live imaging of apical biosynthetic trafficking reveals a post-Golgi sorting role of apical endosomal intermediates. Proc Natl Acad Sci U S A 111:4127-32
Sung, Ching-Hwa; Leroux, Michel R (2013) The roles of evolutionarily conserved functional modules in cilia-related trafficking. Nat Cell Biol 15:1387-97

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