Thesis Title

Characterization and Identification of Putative Collagen Iv Cell Surface Receptors in C. Elegans Through the Utilization of Rna Interference and Bioinformatics

Date of Graduation

Fall 2005

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Colette Witkowski

Keywords

collagen IV, cell surface receptor, RNAi, bioinformatics, microinjection

Subject Categories

Medical Molecular Biology

Abstract

A basement membrane is a thin sheet of extracellular matrix composed of collagen IV, laminin, proteoglycans and nidogen (entactin). The interaction of collagen IV with basement membrane components is required for the integrity, stability, and functions necessary for survival during development. The human genetic diseases Alport’s Syndrome, caused by collagen mutations, and Goodpasture’s Syndrome, a collagen IV targeted autoimmune disease, have demonstrated the importance of a functional collagen IV for normal protein-protein interactions. The current research characterized the three genes, C19H7.1, M01E10.2, and F31F7.2 as putative collagen IV cell surface receptors based on similarity to integrin profiles. The standard non-heritable RNAi delivery system of microinjection was utilized. RNAi acts at the posttranscriptional level down regulating the activity of individual genes. This systemic effect occurs in other tissues and the progeny. An embryonic lethal phenocopy for the ESTs yk557b9.5, yk300c7.5, and yk311g7.5 genes C18H7.1, M01E10.2, and F31F7.2, respectively would indicate the down regulation of a collagen IV receptor; however a WT phenocopy was observed. Information based on bioinformatics databases such as BLAST, File Annotation, the C. elegans database, and recently published literature searches were utilized to identify other genes as putative collagen IV cell surface receptors. Hypothetical C. elegans proteins K07E12.1, Y71F9B.14, and CE02390, were found to have potential domains that could cross the membrane to act as a cell surface receptor and recognize collagen IV.

Copyright

© Shannon M. Burcks

Citation-only

Dissertation/Thesis

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