Masters Thesis

Identification of two uncharacterized proteins that physically interact with the Caenorhabditis elegans SPE-26 protein

The Caenorhabditis elegans SPE-26 protein is involved in the formation of viable spermatids. Two-hybrid interaction analysis was performed in order to determine with which proteins SPE-26 physically interacts. Fusing the carboxy terminal 471 amino acids of the SPE-26 protein to the Escherichia coli LexA protein created a bait protein. This bait was then used to screen a C. elegans cDNA library for potential protein-protein interactions. This library consisted of C. elegans cDNA clones fused to the transcription activator, B42, an E. coli gene that produces a protein that has transcription activating activity. Potential physical interactions were uncovered with two different uncharacterized C. elegans proteins, the collagen-like protein F26B1.4 and the DNAJ-like protein F39B2.10. DNAJ proteins from mice, humans, and rats have been found to be expressed in germ cells, while some collagen proteins are believed to be involved in C. elegans fertility.