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Insulator Protein CP190 Regulates Expression оf Spermatocyte Differentiation Genes in Drosophila melanogaster Male Germline

  • MOLECULAR BIOLOGY OF THE CELL
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Abstract

CP190 protein is one of the key components of Drosophila insulator complexes, and its study is important for understanding the mechanisms of gene regulation during cell differentiation. However, Cp190 mutants die before reaching adulthood, which significantly complicates the study of its functions in imago. To overcome this problem and to investigate the regulatory effects of CP190 in adult tissues development, we have designed a conditional rescue system for Cp190 mutants. Using Cre/loxP-mediated recombination, the rescue construct containing Cp190 coding sequence is effectively eliminated specifically in spermatocytes, allowing us to study the effect of the mutation in male germ cells. Using high-throughput transcriptome analysis we determined the function of CP190 on gene expression in germline cells. Cp190 mutation was found to have opposite effects on tissue-specific genes, which expression is repressed by CP190, and housekeeping genes, that require CP190 for activation. Mutation of Cp190 also promoted expression of a set of spermatocyte differentiation genes that are regulated by tMAC transcriptional complex. Our results indicate that the main function of CP190 in the process of spermatogenesis is the coordination of interactions between differentiation genes and their specific transcriptional activators.

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ACKNOWLEDGMENTS

Authors would like to acknowledge A.K. Golovnin for the provided antibodies (IGB RAS), A.V. Pindurin (IMCB SB RAS) for assistance in cloning, and the “Molecular and Cellular Biology” core facility for the provided equipment.

Funding

The reported study was funded by Russian Foundation for Basic Research, project nos. 19-34-90108, 17-00-00181, and 19-04-00872, and supported by Basic Science Research Program FWGZ-2021-0017 (122011900429-5).

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  1. Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences, 630090, Novosibirsk, Russia

    S. E. Romanov, V. V. Shloma, D. E. Koryakov, S. N. Belyakin & P. P. Laktionov

  2. Novosibirsk State University, 630090, Novosibirsk, Russia

    S. E. Romanov & P. P. Laktionov

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  1. S. E. Romanov
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Correspondence to P. P. Laktionov.

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Statement on the welfare of animals. The conditions in which Drosophila melanogaster was kept complied with the standards specified in Order of the Ministry of Health of Russia no. 267 of June 19, 2003 “On Approval of the Rules of Laboratory Practice in the Russian Federation.”

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Romanov, S.E., Shloma, V.V., Koryakov, D.E. et al. Insulator Protein CP190 Regulates Expression оf Spermatocyte Differentiation Genes in Drosophila melanogaster Male Germline. Mol Biol 57, 113–126 (2023). https://doi.org/10.1134/S0026893323010120

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