dc.contributor.advisor | Buechner, Matthew J. | |
dc.contributor.author | Tong, Xiangyan | |
dc.date.accessioned | 2008-09-07T23:58:08Z | |
dc.date.available | 2008-09-07T23:58:08Z | |
dc.date.issued | 2007-10-12 | |
dc.date.submitted | 2007 | |
dc.identifier.other | http://dissertations.umi.com/ku:2199 | |
dc.identifier.uri | http://hdl.handle.net/1808/4113 | |
dc.description.abstract | Forming and maintaining tubular structure is fundamental to organismal development. The excretory canal cell of C.elegans forms a single-cell epithelial tubule, which provides a simple model for tubular structure study. The EXC proteins regulate maintenance of the apical (lumenal) cytoskeleton of the excretory canal. Loss of exc gene function allows formation of fluid-filled cysts in the excretory canal. exc-9 mutants exhibit short and cystic canals compared to wild-type worms. exc-9 mutants also exhibit tail defects in hermaphrodites and ray defects in male. By SNP mapping, cosmid rescue, and RNAi experiments, we proved that F20D12.5 encodes exc-9. EXC-9 is a homologue of the mammalian intestinal LIM-domain protein CRIP. exc-9 is highly expressed in the canal and tailspike, it is also expressed in some other cells, including UTSE, DTCs, and ALM neurons. Promoter regions important for exc-9 expression were studied. It was found that EXC-9 functions cell-autonomously and the free N-terminus of the protein is required for unextended canal phenotype. Overexpression of exc-9 constructs in an N2 background sometimes causes an "unextended canal" phenotype, in which the canal forms a large cell body filled with lumen with proper diameter, but has no canals along the length of the animal. Since canal extension is sensitive to expression levels of exc-9, injection of the construct also caused unextended canal phenotype in exc-9 mutants. Similar unextended canal phenotype was also found in animals showing high levels of exc-5 expression. I used the unextended canal phenotype to examine epistasis of EXC-9 function with that of other EXC proteins. EXC-9 appears to function upstream of EXC-5 to regulate cytoskeletal formation at the apical surface (possibly via CDC-42); and EXC-9 in turn may depend upon EXC-2 and EXC-4 function. A second well conserved CRIP homologue B0496.7 is highly expressed in multiple valves of C.elegans. It functions similarly to EXC-9 when ectopically expressed in the excretory canal. Expression of the mouse homologue CRIP in the canal failed to rescue exc-9 mutants, but with some modifications, mouse CRIP can function similar as EXC-9. Preliminary data of cloning of exc-2 is reported in this work too. | |
dc.format.extent | 160 pages | |
dc.language.iso | EN | |
dc.publisher | University of Kansas | |
dc.rights | This item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author. | |
dc.subject | Molecular biology | |
dc.title | Cloning and characterization of exc-9, a Caenorhabditis elegans CRIP homologue that regulates tubular structure | |
dc.type | Dissertation | |
dc.contributor.cmtemember | Corbin, Victoria L. | |
dc.contributor.cmtemember | Dentler, William | |
dc.contributor.cmtemember | Lundquist, Erik A. | |
dc.contributor.cmtemember | Timmons, Lisa | |
dc.contributor.cmtemember | Siahaan, Teruna | |
dc.thesis.degreeDiscipline | Biochemistry & Molecular Biology | |
dc.thesis.degreeLevel | PH.D. | |
kusw.oastatus | na | |
kusw.oapolicy | This item does not meet KU Open Access policy criteria. | |
kusw.bibid | 6599220 | |
dc.rights.accessrights | openAccess | |