The Drosophila nuclear receptors EcR and ERR jointly regulate the expression of genes involved in carbohydrate metabolism

Kovalenko, Elena V.; Mazina, M. Yu.; Краснов, А. Н.; Vorobyeva, Nadezhda E.

doi:10.1016/j.ibmb.2019.103184

Cited by 32 publications

(36 citation statements)

References 29 publications

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“…Structurally, BdERR was similar to the estrogen-related receptors (ERRs) of mammals, which indicated that ERRs have highly conserved structural characteristics. A very recent study suggested that the DmEcR and DmERR jointly regulated the expression of glucose metabolism-related genes [35], but their specific functions in B. dorsalis are not yet clear. Our qRT-PCR analysis showed that BdERR was expressed in all development stages, but no obviously high expression was detected (Figure 3).…”

Section: Nr3 Subfamilymentioning

confidence: 99%

Molecular Characterization and Expression Profiling of Nuclear Receptor Gene Families in Oriental Fruit Fly, Bactrocera Dorsalis (Hendel)

Yang

Chen

Song

et al. 2020

Insects

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The oriental fruit fly (Bactrocera dorsalis) is a pest that causes large economic losses in the fruit and vegetable industry, so its control is a major challenge. Nuclear receptors (NRs) are a superfamily of ligand-dependent transcription factors that directly combine with DNA to regulate the expression of downstream target genes. NRs are closely associated with multiple physiological processes such as metabolism, reproduction, and development. Through sequence searches and analysis, we identified 21 B. dorsalis NR genes, all of which contained at least one of the two characteristic binding domains. On the basis of the conserved sequences and phylogenetic relationships, we divided the 21 NR genes into seven subfamilies. All members of the NR0 subfamily and BdHR83, which belonged to the NR2E group, lacked ligand-binding domains. The BdDSF and BdHR51, which also belonged to the NR2Egroup, and BdE78 (which belonged to the NR1E group) all lacked DNA-binding domains. The BdDSF and BdHR83 sequences were incomplete, and were not successfully amplified. Development-and tissue-specific expression profiling demonstrated that the transcript levels of the 19 NR genes varied considerably among eggs, larva, pupae, and adults, as well as among larval and adult male and female tissues. Our results will contribute to a better understanding of NR evolution and expand our knowledge of B. dorsalis physiology.Insects 2020, 11, 126 2 of 15 in recognizing hormone response elements. The D domain functions as a hinge between the DBD and the ligand-binding domain (LBD). The LBD (also known as E domain) contains a hydrophobic ligand-binding pocket that is specific to each receptor and is the source of the sequence variability within the LBD. In addition, the LBD domain mediates dimerization and the ligand-dependent transcriptional activation function (AF-2) [4]. The C-terminal contains the F domain, which is not present in all NRs and is highly diverse.A large number of NR gene sets have been characterized from the whole-genome sequence of many animals. For example, 48 NRs were identified in the human genome [5], over 270 NRs have been found in Caenorhabditis elegans [6], and 49 and 47 were found in mouse and rat, respectively [7]. Surprisingly, only 21 NRs were found in Drosophila melanogaster [8], 19 in Bombyx mori [9], 22 in Apis mellifera [10], and 21 in Tribolium castaneum [11].In D. melanogaster, seven NR genes that are regulated directly by 20E have been identified, namely EcR, E75, E78, FTZ-F1, HR3, HR4, and HR39 [12]. Among them, E75 is the primary response gene of 20E. The others are secondary response genes that are maximally expressed after 20E-induced protein synthesis has begun [13]. The ligand-receptor complex of 20E-EcR-USP induces the expression of the 20E primary response gene. Then, the cascade of 20E secondary response genes, which is induced by the transcription factor encoded by the 20E primary response gene, amplifies the 20E signal by regulating the expression of the secondary response genes (e.g., HR3, HR4, and FTZ...

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Section: Nr3 Subfamilymentioning

confidence: 99%

Molecular Characterization and Expression Profiling of Nuclear Receptor Gene Families in Oriental Fruit Fly, Bactrocera Dorsalis (Hendel)

Yang

Chen

Song

et al. 2020

Insects

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“…The ultimate goal is to understand the general mechanism for most genes, including the cooperative function of hundreds of transcriptional factors. As an experimental approach we use Drosophila melanogaster developmental genes (e.g., 20-hydroxyecdysone (20E)-dependent), which are a well-regulated physiological system that is convenient to study 11,12 . The main molecular sensor of an ecdysone system is the EcR/Usp heterodimer.…”

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confidence: 99%

“…Polyclonal cell lines bearing stably integrated expression constructs were selected and maintained subsequently in Schneider medium supplemented with Blasticidin (Sigma-Aldrich) at a final concentration of 20 μg/mL. Immunoprecipitation 11 and RNA interference-mediated knockdowns of CP190, Chro, Mor, Brm, Mi-2, NELF A, Spt5 and Nup358 in Drosophila S2 cells 15 were performed as previously described. Efficiency of RNA knockdown was evaluated by qPCR and provided at Fig.…”

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confidence: 99%

Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner

Mazina

Ziganshin

Magnitov

et al. 2020

Sci Rep

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proximity-dependent biotin labelling revealed undescribed participants of the ecdysone response in Drosophila. Two labelling enzymes (BioID2 and APEX2) were fused to EcR or Usp to biotin label the surrounding proteins. the ecR/Usp heterodimer was found to collaborate with nuclear pore subunits, chromatin remodelers, and architectural proteins. Many proteins identified through proximity-dependent labelling with ecR/Usp were described previously as functional components of an ecdysone response, corroborating the potency of this labelling method. A link to ecdysone response was confirmed for some newly discovered regulators by immunoprecipitation of prepupal nuclear extract with anti-ecR antibodies and functional experiments in Drosophila S2 cells. A more in-depth study was conducted to clarify the association of EcR/Usp with one of the detected proteins, CP190, a well-described cofactor of Drosophila insulators. CP190 was found to co-immunoprecipitate with the EcR subunit of EcR/Usp in a 20E-independent manner. ChIP-Seq experiments revealed only partial overlapping between CP190 and EcR bound sites in the Drosophila genome and complete absence of CP190 binding at 20E-dependent enhancers. Analysis of Hi-C data demonstrated an existence of remote interactions between 20E-dependent enhancers and CP190 sites which suggests formation of a protein complex between EcR/Usp and CP190 through the space. Our results support the previous concept that CP190 has a role in stabilization of specific chromatin loops for proper activation of transcription of genes regulated by 20E hormone.www.nature.com/scientificreports www.nature.com/scientificreports/ rate (e.g., promoter-bound transcriptional complexes). Recently, an additional directed mutagenesis procedure of BirA has allowed for the development of the fast TurboID enzyme 7 .An alternative to the BioID2 technology is a combination of APEX2, a plant ascorbate peroxidase, and a biotin-phenol substrate. In the presence of hydrogen peroxide, the APEX2 enzyme can catalyse the substrate conversion (biotin-phenol) into active and short-lived radicals which can modify the surrounding proteins 3 . Although the method is less common than the BioID technique, APEX2 labelling requires significantly less time to biotinylate surrounding proteins, making it more suited for researching unstable complexes 8,9 . For example, it was recently integrated with the CRISPR/Cas9 system to investigate DNA-protein interactions, as an alternative to ChIP-Seq 10 .The main research interest of our group is to accurately describe the molecular mechanisms of transcriptional activation in eukaryotes. The ultimate goal is to understand the general mechanism for most genes, including the cooperative function of hundreds of transcriptional factors. As an experimental approach we use Drosophila melanogaster developmental genes (e.g., 20-hydroxyecdysone (20E)-dependent), which are a well-regulated physiological system that is convenient to study 11,12 . The main molecular sensor of an ecdysone system is the EcR/Us...

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“…The 20E led to cell cycle arrest in the G2/M cell phase through the EcR receptor while inhibiting DNA replication. In general, 20E functions by activating the ecdysone receptor (EcR) to regulate the expression of specific genes [ 11 ]. Flow cytometry and the BrdU assay demonstrated that knocking down the receptor EcR can eliminate the effect of 20E.…”

Section: Discussionmentioning

confidence: 99%

“…The PG secretes ecdysone, which is converted to 20E that is active in the peripheral tissues including the fat body, midgut, and muscle [ 10 ]. Ecdysteroids generally function by activating the ecdysone receptor (EcR), a member of the transcription factor nuclear receptor family, to regulate the expression of specific genes [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

BmFoxO Gene Regulation of the Cell Cycle Induced by 20-Hydroxyecdysone in BmN-SWU1 Cells

Zhang

Yang

Chen

et al. 2020

Insects

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Ecdysteroid titer determines the state of the cell cycle in silkworm (Bombyxmori) metamorphosis. However, the mechanism of this process is unclear. In this study, we demonstrated that the BmFoxO gene participates in the regulation of the cell cycle induced by 20-Hydroxyecdysone (20E) in BmN-SWU1 cells. The 20E blocks the cell cycle in the G2/M phase through the ecdysone receptor (EcR) and inhibits DNA replication. The 20E can promote BmFoxO gene expression. Immunofluorescence and Western blot results indicated that 20E can induce BmFoxO nuclear translocation in BmN-SWU1 cells. Overexpression of the BmFoxO gene affects cell cycle progression, which results in cell cycle arrest in the G0/G1 phase as well as inhibition of DNA replication. Knockdown of the BmFoxO gene led to cell accumulation at the G2/M phase. The effect of 20E was attenuated after BmFoxO gene knockdown. These findings increase our understanding of the function of 20E in the regulation of the cell cycle in B. mori.

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The Drosophila nuclear receptors EcR and ERR jointly regulate the expression of genes involved in carbohydrate metabolism

Cited by 32 publications

References 29 publications

Molecular Characterization and Expression Profiling of Nuclear Receptor Gene Families in Oriental Fruit Fly, Bactrocera Dorsalis (Hendel)

Molecular Characterization and Expression Profiling of Nuclear Receptor Gene Families in Oriental Fruit Fly, Bactrocera Dorsalis (Hendel)

Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner

BmFoxO Gene Regulation of the Cell Cycle Induced by 20-Hydroxyecdysone in BmN-SWU1 Cells

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