[
Mid-west Worm Meeting,
2002]
The 26S proteasome is a large protein complex that is responsible for the degradation of cellular proteins. The ubiquitin-proteasome pathway is the primary means of protein degradation. Our lab is using RNA-mediated-interference (RNAi) to determine the function of the regulatory particle protein subunits within the 26S proteasome. The 26S proteasome consists of a core particle (alpha- and beta- subunits) and either 19S or 11S particles flanking one or both ends of the core particle. The catalytic portion of the proteasome is within the core particle. RNAi of five gene products corresponding to a respective core particle subunit resulted in 100% lethality. The 19S regulatory particle consists of 6 ATPase subunits (RPT's) and 12-13 non-ATPase subunits (RPN's). Using Wormbase, a list of C. elegans ORF's corresponding to yeast regulatory particle subunits was created. A total of 19 putative genes were found including three RPT6's and two RPN2 splice variants. RNAi has been performed on 15 of the 19 genes. Seven of the 15 showed significant embryonic lethality. Continued investigation will include combinatorial RNAi on the three RPT6's and all the subunits that showed no significant embryonic lethality. Phenotype analysis of embryos is underway.