Transgenic C. elegans dauer larvae expressing hookworm phospho null DAF-16/FoxO exit dauer

Verena Gelmedin, Thomas Brodigan, Xin Gao, Michael Krause, Zhu Wang, John M. Hawdon

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Parasitic hookworms and the free-living model nematode Caenorhabtidis elegans share a developmental arrested stage, called the dauer stage in C. elegans and the infective third-stage larva (L3) in hookworms. One of the key transcription factors that regulate entrance to and exit from developmental arrest is the forkhead transcription factor DAF-16/FoxO. During the dauer stage, DAF-16 is activated and localized in the nucleus. DAF-16 is negatively regulated by phosphorylation by the upstream kinase AKT, which causes DAF-16 to localize out of the nucleus and the worm to exit from dauer. DAF-16 is conserved in hookworms, and hypothesized to control recovery from L3 arrest during infection. Lacking reverse genetic techniques for use in hookworms, we used C. elegans complementation assays to investigate the function of Ancylostoma caninum DAF-16 during entrance and exit from L3 developmental arrest. We performed dauer switching assays and observed the restoration of the dauer phenotype when Ac-DAF-16 was expressed in temperature-sensitive dauer defective C. elegans daf-2(e1370);daf-16(mu86) mutants. AKT phosphorylation site mutants of Ac-DAF-16 were also able to restore the dauer phenotype, but surprisingly allowed dauer exit when temperatures were lowered. We used fluorescence microscopy to localize DAF-16 during dauer and exit from dauer in C. elegans DAF-16 mutant worms expressing Ac-DAF-16, and found that Ac-DAF-16 exited the nucleus during dauer exit. Surprisingly, Ac-DAF-16 with mutated AKT phosphorylation sites also exited the nucleus during dauer exit. Our results suggest that another mechanism may be involved in the regulation DAF-16 nuclear localization during recovery from developmental arrest.

Original languageEnglish (US)
Article numbere25996
JournalPLoS One
Volume6
Issue number10
DOIs
StatePublished - Oct 7 2011

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Ancylostomatoidea
hookworms
Phosphorylation
nematode larvae
Larva
genetically modified organisms
phosphorylation
Assays
mutants
Forkhead Transcription Factors
Recovery
transcription factors
Fluorescence microscopy
Ancylostoma
Ancylostoma caninum
Phenotype
Genetic Techniques
phenotype
Reverse Genetics
Restoration

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Transgenic C. elegans dauer larvae expressing hookworm phospho null DAF-16/FoxO exit dauer. / Gelmedin, Verena; Brodigan, Thomas; Gao, Xin; Krause, Michael; Wang, Zhu; Hawdon, John M.

In: PLoS One, Vol. 6, No. 10, e25996, 07.10.2011.

Research output: Contribution to journalArticle

Gelmedin, Verena ; Brodigan, Thomas ; Gao, Xin ; Krause, Michael ; Wang, Zhu ; Hawdon, John M. / Transgenic C. elegans dauer larvae expressing hookworm phospho null DAF-16/FoxO exit dauer. In: PLoS One. 2011 ; Vol. 6, No. 10.
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