Oxidative stress responses in Caenorhabditis elegans with reduced mrck‐1 expression

Abstract

MRCK‐1 (myotonic dystrophy kinase‐related Cdc42 binding kinase) acts downstream of the Rho GTPase CDC‐42 to determine cell polarity and is critical for C. elegans embryonic development. Given the conserved GXXXXCGKTC redox‐sensitive motif in the phosphoryl‐binding loop of CDC‐42, a role for MRCK‐1 in cellular responses to reactive oxygen species (ROS) is being explored. An RNA interference (RNAi) construct of 190 bp targeting exon 5 of the mrck‐1 gene on chromosome V was cloned into the RNAi expression vector pL4440 to enable production of double‐stranded RNA. Mrck‐1 RNAi induction in worm strains with oxidative stress responsive green fluorescent protein (GFP)‐ promoter fusions, including hsp‐16.2, sod‐3 and gst‐4, enables the measurement of oxidative stress induced GFP expression. Ongoing experiments include exposing worm strains to temperature, ultra‐violet (UV) light and chemical stressors to elucidate a possible phenotype related to diminished MRCK‐1 function under oxidative stress conditions. This work was supported by the Henson Undergraduate Research Grant program.