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IDENTIFICATION OF NOVEL PROTEIN-PROTEIN INTERACTIONS
AND FUNCTIONAL ANALYSIS IN THE MOUSE PHOTORECEPTOR
OF THE HYPOTHETICAL PROTEIN FLJ33282-ALS2CR4
AND THE SMALL RHO GTPASE, RND2
by
Freddi Isaac Zuniga
A Dissertation Presented to the
FACULTY OF THE USC GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
DOCTOR OF PHILOSOPHY
(GENETIC, MOLECULAR and CELLULAR BIOLOGY)
August 2010
Copyright 2010 Freddi Isaac Zuniga
Object Description
| Title | Identification of novel protein-protein interactions and functional analysis in the mouse photoreceptor of the hypothetical protein FLJ33282-Als2cr4 and the small Rho GTPase, Rnd2 |
| Author | Zuniga, Freddi Isaac |
| Author email | fzuniga@usc.edu; fzuniga.usc@gmail.com |
| Degree | Doctor of Philosophy |
| Document type | Dissertation |
| Degree program | Genetic, Molecular & Cellular Biology |
| School | Keck School of Medicine |
| Date defended/completed | 2010-06-07 |
| Date submitted | 2010 |
| Restricted until | Unrestricted |
| Date published | 2010-06-28 |
| Advisor (committee chair) | Craft, Cheryl M. |
| Advisor (committee member) |
Sampath, Alapakkam P. Chen, Jeannie Fong, Henry K. W. Hinton, David R. |
| Abstract | The main focus of this research project was to decipher the role of the hypothetical protein FLJ33282-Als2cr4 (amyotrophic lateral sclerosis 2 [juvenile] chromosome region, candidate 4; NM_001037812) in the mouse retina. We characterized its molecular structure, cellular interacting partners and potential biochemical functions. Previous in situ hybridization and gene expression profiles showed mRNAs encoding Als2cr4 are abundant in the eye, hippocampus, cerebellum, and olfactory bulb from embryonic day 15 and throughout adulthood in mouse.; Antibodies were developed from predicted antigenic epitopes of Als2cr4, characterized, and used to examine protein expression and morphological localization in retinas from light-adapted and dark-adapted mice with immunohistochemistry, immunoblot analysis and immuno-electron microscopy, followed with immunoprecipitation studies to identify potential functional interacting proteins by mass spectroscopy analysis.; My doctoral work demonstrates that Als2cr4 is a 45 kDa insoluble protein, which is highly enriched in mouse retina and localizes to photoreceptor outer segments, ciliary complex, and outer plexiform layer. The immuno-electron microscopy for Als2cr4 verifies its expression pattern in the discs of photoreceptor outer segments. Furthermore, immunoprecipitation analysis followed by mass spectroscopy identified eight novel interacting partners.; Our data provides evidence that the novel protein Als2cr4 has a predicted tetraspanin-like membrane structure, is localized to photoreceptors and to the post-synaptic outer plexiform layer, and interacts with cytoskeletal proteins. We predict that Als2cr4 is involved in membrane transport between the photoreceptor inner segment and outer segment and is a key component in maintaining outer segment structural integrity. |
| Keyword | cytoskeleton; ophthalmology; photoreceptor; structure |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Provenance | Electronically uploaded by the author |
| Type | texts |
| Legacy record ID | usctheses-m3160 |
| Rights | Zuniga, Freddi Isaac |
| Repository name | Libraries, University of Southern California |
| Repository address | Los Angeles, California |
| Repository email | http://www.usc.edu/isd/libraries/services/ask_a_librarian/email/ |
| Filename | etd-Zuniga-3897 |
| Archival file | uscthesesreloadpub_Volume26/etd-Zuniga-3897.pdf |
Description
| Title | Page 1 |
| Full text | IDENTIFICATION OF NOVEL PROTEIN-PROTEIN INTERACTIONS AND FUNCTIONAL ANALYSIS IN THE MOUSE PHOTORECEPTOR OF THE HYPOTHETICAL PROTEIN FLJ33282-ALS2CR4 AND THE SMALL RHO GTPASE, RND2 by Freddi Isaac Zuniga A Dissertation Presented to the FACULTY OF THE USC GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY (GENETIC, MOLECULAR and CELLULAR BIOLOGY) August 2010 Copyright 2010 Freddi Isaac Zuniga |
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