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Proc Natl Acad Sci U S A. 2016 Feb 9;113(6):E735-43. doi: 10.1073/pnas.1523838113. Epub 2016 Jan 7.

Hairy and Groucho mediate the action of juvenile hormone receptor Methoprene-tolerant in gene repression.

Author information

1
Department of Entomology, University of California, Riverside, CA 92521; The Institute for Integrative Genome Biology, University of California, Riverside, CA 92521;
2
Department of Entomology, University of California, Riverside, CA 92521;
3
Department of Entomology, University of California, Riverside, CA 92521; College of Plant Protection, Southwest University, Chongqing 400715, People's Republic of China;
4
State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China;
5
The Institute for Integrative Genome Biology, University of California, Riverside, CA 92521; Department of Botany and Plant Sciences, University of California, Riverside, CA 92521.
6
Department of Entomology, University of California, Riverside, CA 92521; The Institute for Integrative Genome Biology, University of California, Riverside, CA 92521; alexander.raikhel@ucr.edu.

Abstract

The arthropod-specific juvenile hormone (JH) controls numerous essential functions. Its involvement in gene activation is known to be mediated by the transcription factor Methoprene-tolerant (Met), which turns on JH-controlled genes by directly binding to E-box-like motifs in their regulatory regions. However, it remains unclear how JH represses genes. We used the Aedes aegypti female mosquito, in which JH is necessary for reproductive maturation, to show that a repressor, Hairy, is required for the gene-repressive action of JH and Met. The RNA interference (RNAi) screen for Met and Hairy in the Aedes female fat body revealed a large cohort of Met- and Hairy-corepressed genes. Analysis of selected genes from this cohort demonstrated that they are repressed by JH, but RNAi of either Met or Hairy renders JH ineffective in repressing these genes in an in vitro fat-body culture assay. Moreover, this JH action was prevented by the addition of the translational inhibitor cycloheximide (CHX) to the culture, indicating the existence of an indirect regulatory hierarchy. The lack of Hairy protein in the CHX-treated tissue was verified using immunoblot analysis, and the upstream regions of Met/Hairy-corepressed genes were shown to contain common binding motifs that interact with Hairy. Groucho (gro) RNAi silencing phenocopied the effect of Hairy RNAi knockdown, indicating that it is involved in the JH/Met/Hairy hierarchy. Finally, the requirement of Hairy and Gro for gene repression was confirmed in a cell transfection assay. Thus, our study has established that Hairy and its cofactor Gro mediate the repressive function of JH and Met.

KEYWORDS:

groucho; hairy; juvenile hormone; receptor; repressor

PMID:
26744312
PMCID:
PMC4760797
DOI:
10.1073/pnas.1523838113
[Indexed for MEDLINE]
Free PMC Article

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