Heat Shock Protein 83 (Hsp83) Facilitates Methoprene-tolerant (Met) Nuclear Import to Modulate Juvenile Hormone Signaling

He, Qianyu; Wen, Di; Jia, Qiangqiang; Cui, Chunlai; Wang, Jian; Palli, Subba Reddy; Li, Sheng

doi:10.1074/jbc.m114.582825

Cited by 76 publications

(110 citation statements)

References 45 publications

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“…After extensive washing with cold Nonidet P-40 buffer, the samples were treated with RIPA lysis buffer (Beyotime) about 15 min on the ice. Then immunoprecipitates were separated by SDS-PAGE and analyzed by Western blots (35).…”

Section: Methodsmentioning

confidence: 99%

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20-Hydroxyecdysone (20E) Primary Response Gene E93 Modulates 20E Signaling to Promote Bombyx Larval-Pupal Metamorphosis

Liu

Dai

Guo

et al. 2015

Journal of Biological Chemistry

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105

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Background: 20-Hydroxyecdysone (20E) and juvenile hormone (JH) coordinately control insect molting and metamorphosis. Results: Induced by 20E and suppressed by JH, E93 acts through GAGA-containing motifs and up-regulates a subset of 20E response genes. Conclusion: E93 transcriptionally modulates 20E signaling to promote Bombyx metamorphosis. Significance: Our study helps in understanding the complicated molecular mechanisms of insect metamorphosis.

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Section: Methodsmentioning

confidence: 99%

“…The relative luciferase activity was calculated by normalizing the reporter firefly luciferase level to the reference Renilla luciferase level. Dual luciferase assays were conducted using the Dual Luciferase Assay System (Promega) and a Modulus Luminometer (Turner BioSystems) (24,25,32,35).…”

Section: Methodsmentioning

confidence: 99%

20-Hydroxyecdysone (20E) Primary Response Gene E93 Modulates 20E Signaling to Promote Bombyx Larval-Pupal Metamorphosis

Liu

Dai

Guo

et al. 2015

Journal of Biological Chemistry

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105

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“…Bm-N cells were grown in 10-cm dishes (70% confluent) and transfected with the E75A/B/C-V5 expression plasmid for 48 h using the Effectene transfection reagent (Qiagen, Germany). Then, the cells were fixed and subjected to ChIP assay (31,42,43) using the agarose ChIP kit (Pierce) and the V5 antibody (Sigma). Mock immunoprecipitations with pre-immune serum were used for negative controls.…”

Section: Methodsmentioning

confidence: 99%

“…The relative luciferase activity was calculated by normalizing the reporter firefly luciferase level to the reference Renilla luciferase level. Dual-luciferase assays were conducted using the dual-luciferase assay system (Promega) and a Modulus luminometer (Turner BioSystems) (29,31,42,43). For some experiments, two constructs of EGFP, E75A, E75B, E75C, or coE75A/B/C were co-transfected into HEK 293 cells equally or in a dose-dependent manner.…”

Section: Methodsmentioning

confidence: 99%

“…After 1 day of pre-incubation, the cells were co-transfected with two pcDNA 3.1(ϩ) vectors expressing E75A, E75B (or its mutants coE75A/C, coE75A/B/C, and E75noN), or E75C, the C termini of which were fused to different tags (V5, HA, FLAG, and Myc), for 48 h. After extensive washing, the coverslips were fixed in 4% paraformaldehyde for 45 min at room temperature, blocked in phosphate-buffered saline containing 5% BSA and 1% Triton X-100 (PBSBT) for 1 h, and incubated with two different primary tag antibodies (V5, HA, FLAG, and Myc, Sigma) (diluted 1:200) at 4°C overnight. The coverslips were washed for 1 h in PBSBT and incubated with two counterpart FITC green/redconjugated secondary antibodies from mouse/rabbit (diluted 1:200) for 2 h at room temperature (43). Images were captured using the Olympus FV10-ASW confocal microscope at ϫ40 magnification.…”

Section: Methodsmentioning

confidence: 99%

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20-Hydroxyecdysone (20E) Primary Response Gene E75 Isoforms Mediate Steroidogenesis Autoregulation and Regulate Developmental Timing in Bombyx

Tian

Guo

et al. 2016

Journal of Biological Chemistry

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The temporal control mechanisms that precisely control animal development remain largely elusive. The timing of major developmental transitions in insects, including molting and metamorphosis, is coordinated by the steroid hormone 20-hydroxyecdysone (20E). 20E involves feedback loops to maintain pulses of ecdysteroid biosynthesis leading to its upsurge, whereas the underpinning molecular mechanisms are not well understood. Using the silkworm Bombyx mori as a model, we demonstrated that E75, the 20E primary response gene, mediates a regulatory loop between ecdysteroid biosynthesis and 20E signaling. E75 isoforms A and C directly bind to retinoic acid receptor-related response elements in Halloween gene promoter regions to induce gene expression thus promoting ecdysteroid biosynthesis and developmental transition, whereas isoform B antagonizes the transcriptional activity of isoform A/C through physical interaction. As the expression of E75 isoforms is differentially induced by 20E, the E75-mediated regulatory loop represents a fine autoregulation of steroidogenesis, which contributes to the precise control of developmental timing.Animals undergo developmental transitions from the embryo to juvenile to adulthood, and these processes are determined by steroid hormones and their corresponding nuclear receptors (NRs).2 In insects, 20-hydroxyecdysone (20E; ecdysone is the immediate precursor of 20E; 20E and ecdysone are the main ecdysteroids) is the actual steroid hormone. The ecdysone receptor (EcR) and its partner molecule, Ultraspiracle (USP), form the functional NR complex of 20E. In conjunction with EcR-USP, 20E activates a small set of early response genes encoding several transcription factors that further activate a large set of downstream late response genes. Pulses of 20E signals initiate major developmental transitions in insects, including egg hatching, larval-larval molting, and larval-pupal-adult metamorphosis (1, 2). NRs form a large and conserved superfamily of ligand-activated transcription factors that are essential for growth, development, reproduction, homeostasis, and metabolism. NRs are defined by the presence of a highly conserved DNA binding domain (DBD) and a less conserved ligand binding domain (3, 4). There are 18 -19 NRs in insects, including the fruit fly, Drosophila melanogaster, and the silkworm, Bombyx mori (3,5,6). Apart from the EcR, ligand was only identified for another insect NR, ecdysone-induced protein 75B (E75). E75 is a crucial 20E response gene that affects ecdysteroid titer. E75 binds to heme, which responds to gases . The E75 orthologs in mammals are Rev-erb ␣ (NR1D1) and Rev-erb ␤ (NR1D2), and NR1D2 binds to heme, responds to NO, and regulates circadian rhythm (13-15).In Drosophila, the E75 locus encodes four E75 mRNA isoforms, E75A, E75B, E75C, and E75D, which are generated by differential promoter usage and alternative splicing of 5Ј exons. The DBD of E75A/C possesses two C4 zinc fingers; E75B is incomplete and contains only one zinc finger, whereas E75D lacks a DBD. 20E-...

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Detection of juvenile hormone agonists by a new reporter gene assay using yeast expressing Drosophila methoprene‐tolerant

et al. 2021

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Juvenile hormones (JHs) are sesquiterpenoids that play important roles in the regulation of growth, metamorphosis, and reproduction in insects. Synthetic JH agonists (JHAs) have been used as insecticides and are categorized as a class of insect growth regulators (IGRs). Natural JHs and synthetic JHAs bind to the JH receptor methoprene‐tolerant (Met), which forms a functional JH‐receptor complex with steroid receptor coactivators, such as Drosophila melanogaster Taiman (Tai). The ligand‐bound Met–Tai complex induces the transcription of JH response genes by binding to specific DNA elements referred to as JH response elements (JHREs). In the present study, we established a reporter gene assay (RGA) for detecting natural JHs and synthetic JHAs in a yeast strain expressing D. melanogaster Met and Tai. The yeast RGA system detected various juvenoid ligands in a dose‐dependent manner. The rank order of the ligand potencies of the juvenoids examined in the yeast RGA linearly correlated with those of RGAs for Met–Tai established in mammalian and insect cells. Our new yeast RGA is rapid, easy to handle, cost‐effective, and valuable for screening novel JHAs.

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Heat Shock Protein 83 (Hsp83) Facilitates Methoprene-tolerant (Met) Nuclear Import to Modulate Juvenile Hormone Signaling

Cited by 76 publications

References 45 publications

20-Hydroxyecdysone (20E) Primary Response Gene E93 Modulates 20E Signaling to Promote Bombyx Larval-Pupal Metamorphosis

20-Hydroxyecdysone (20E) Primary Response Gene E93 Modulates 20E Signaling to Promote Bombyx Larval-Pupal Metamorphosis

20-Hydroxyecdysone (20E) Primary Response Gene E75 Isoforms Mediate Steroidogenesis Autoregulation and Regulate Developmental Timing in Bombyx

Detection of juvenile hormone agonists by a new reporter gene assay using yeast expressing Drosophila methoprene‐tolerant

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