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Oxidative stress responses in Caenorhabditis elegans with reduced mrck‐1 expression
Author(s) -
Rigoulot Stephen,
Blondeaux Sarah,
Russo Matthew,
Erickson Patti T.
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.768.4
Subject(s) - rna interference , caenorhabditis elegans , biology , microbiology and biotechnology , gene knockdown , green fluorescent protein , oxidative stress , small hairpin rna , gene , rna , biochemistry
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.