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Phosphorylation of the Conserved Transcription Factor ATF-7 by PMK-1 p38 MAPK Regulates Innate Immunity in Caenorhabditis elegans

Figure 1

Characterization of atf-7 mutants that affect signaling downstream of PMK-1 p38 MAPK.

(A) Fluorescence microscopy images of GFP expression from the agIs219 transgene in wild-type, atf-7(qd22) and atf-7(qd22 qd130) one-day-old adults. (B) Pathogenesis assay of L4 larval stage wild-type worms, atf-7(qd22) and atf-7(qd22 qd130) mutant animals, on P. aeruginosa PA14. All strains carry the agIs219 transgene. The differences in susceptibility between atf-7(qd22) mutant animals and wild-type worms, atf-7(qd22) and atf-7(qd22 qd130) mutant animals, and atf-7(qd22 qd130) mutant animals and wild-type worms are all significant (p<0.0001 for each comparison). Replicate data can be seen in Figure S10. (C) Immunoblot analysis of worm lysates from atf-7(qd22) worms. Total PMK-1 was identified using a polyclonal antibody generated against C. elegans PMK-1. Activated PMK-1 levels were identified using an antibody specific for the doubly phosphorylated TGY motif of activated PMK-1. (D) Pathogenesis assay of L4 larval stage wild-type worms; atf-7(qd22 qd130) and atf-7(qd137) mutants; and atf-7(qd22 qd130)/atf-7(+), atf-7(qd137)/atf-7(+), and atf-7(qd22 qd130)/atf-7(qd137) trans-heterozygotes, on P. aeruginosa PA14. All strains carry the agIs219 transgene. The differences in susceptibility between atf-7(qd22 qd130) mutant animals and atf-7(qd22 qd130)/atf-7(+) trans-heterozygotes, and atf-7(qd137) mutants animals and atf-7(qd137)/atf-7(+) trans-heterozygotes are significant (p<0.0001 for each comparison). There is no difference in susceptibility between atf-7(qd22 qd130) mutant animals and atf-7(qd22 qd130)/atf-7(qd137) trans-heterozygotes, and atf-7(qd137) mutant animals and atf-7(qd22 qd130)/atf-7(qd137) trans-heterozygotes (p>0.35 for each comparison). Replicate data can be seen in Figure S13.

Figure 1

doi: https://doi.org/10.1371/journal.pgen.1000892.g001