The Ras signalling pathway plays several important roles in the development of the nematode Caenorhabditis elegans. So far, two types of Ras-GTPase activating proteins (Ras-GAPs) have been identified in this organism. To aid the study of the regulation and function of the Ras pathway, we set out to isolate a new GAP gene from C. elegans by transcomplementation of the fission yeast gap1 mutant.
We isolated a C. elegans cDNA that encoded a protein which was similar to, but not exactly homologous with mammalian p120 Ras-GAP. This gene, named gap-2, generated at least nine distinct mRNA species through transcription from different promoters and subsequent alternative splicing involving 25 exons. These isoforms were differentially expressed among tissues. A deletion of gap-2 caused no obvious phenotype by itself, but a loss of gap-2 function could suppress larval lethality in both let-23 and let-60 reduction-of-function mutants, in which the Ras activity was lowered.
C. elegans gap-2 encodes a novel Ras-GAP, which is similar to vertebrate p120 but which may constitute a new GAP subfamily. gap-2 mRNA isoforms arise by an unusually extensive variation in initiation sites and associated alternative splicing, and each isoform may play a distinct role in specific tissues. GAP-2 appears to function as a negative regulator of LET-60 Ras during larval development.