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The Mediator subunit MDT-15 acts downstream of the p38 MAP kinase PMK-1 to regulate the induction of F08G5.6 and F35E12.5.

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posted on 2014-05-29, 04:26 authored by Read Pukkila-Worley, Rhonda L. Feinbaum, Deborah L. McEwan, Annie L. Conery, Frederick M. Ausubel

(A) Wild-type or mdt-15(tm2182) mutant synchronized L1 animals containing the pF08G5.6::GFP immune reporter were grown on vector control (L4440), vhp-1(RNAi) or a combination of vhp-1(RNAi) and pmk-1(RNAi) bacteria and then transferred as L4 animals to PA14 for 18 hours. Animals were photographed under the same imaging conditions. (B) qRT-PCR was used to examine the expression levels of F08G5.6, F35E12.5 and C32H11.1 in wild-type N2 and mdt-15(tm2182) mutant animals exposed to vhp-1(RNAi) or the vector control (L4440) under basal conditions (as described above) and 8 hours after exposure to P. aeruginosa. Knockdown of vhp-1 caused significant induction of F08G5.6 and F35E12.5 in wild-type N2 animals (p<0.001), but not in mdt-15(tm2182) animals (p>0.05), under baseline (E. coli) and pathogen-induced conditions. The expression of C32H11.1 was significantly induced by vhp-1(RNAi) (p<0.001) in an mdt-15-dependent manner under baseline conditions (p<0.001), but not following exposure to P. aeruginosa. Data are the average of two biological replicates each normalized to a control gene with error bars representing SEM and are presented as the value relative to the average expression of the indicated gene in the baseline condition (L4440 animals exposed to E. coli).

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