期刊
BMC BIOLOGY
卷 19, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s12915-021-00952-2
关键词
20-hydroxyecdysone; Krü ppel homolog 1; Kr-h1 regulatory protein; Vitellogenin receptor; Oogenesis
类别
资金
- Guangdong Provincial Natural Science Foundation [2017A030306003]
- Guangdong Provincial Special Support Program [2017TQ04N744]
- Chinese National Natural Science Foundation [31872969, 32000338]
The transcription factor BmKRP was identified as a new mediator of 20E regulation of B. mori oogenesis. The study suggests that induction of BmKRP by 20E regulates BmKr-h1 expression, which then induces BmVgR expression to facilitate Vg uptake and oogenesis. Experimentally blocking BmKRP led to suppressed transcription of BmKr-h1 and BmVgR, resulting in arrested oogenesis.
Background Kruppel homolog 1 (Kr-h1) is a critical transcription factor for juvenile hormone (JH) signaling, known to play a key role in regulating metamorphosis and adult reproduction in insects. Kr-h1 can also be induced by molting hormone 20-hydroxyecdysone (20E), however, the underlying mechanism of 20E-induced Kr-h1 expression remains unclear. In the present study, we investigated the molecular mechanism of Kr-h1 induction by 20E in the reproductive system of a model lepidopteran insect, Bombyx mori. Results Developmental and tissue-specific expression analysis revealed that BmKr-h1 was highly expressed in ovaries during the late pupal and adult stages and the expression was induced by 20E. RNA interference (RNAi)-mediated depletion of BmKr-h1 in female pupae severely repressed the transcription of vitellogenin receptor (VgR), resulting in the reduction in vitellogenin (Vg) deposition in oocytes. BmKr-h1 specifically bound the Kr-h1 binding site (KBS) between - 2818 and - 2805 nt upstream of BmVgR and enhanced the transcription of BmVgR. A 20E cis-regulatory element (CRE) was identified in the promoter of BmKr-h1 and functionally verified using luciferase reporter assay, EMSA and DNA-ChIP. Using pull-down assays, we identified a novel transcription factor B. mori Kr-h1 regulatory protein (BmKRP) that specifically bound the BmKr-h1 CRE and activated its transcription. CRISPR/Cas9-mediated knockout of BmKRP in female pupae suppressed the transcription of BmKr-h1 and BmVgR, resulting in arrested oogenesis. Conclusion We identified BmKRP as a new transcription factor mediating 20E regulation of B. mori oogenesis. Our data suggests that induction of BmKRP by 20E regulates BmKr-h1 expression, which in turn induces BmVgR expression to facilitate Vg uptake and oogenesis.
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