Genetically Derepressed Nucleoplasmic Stellate Protein in Spermatocytes of D. melanogaster Interacts with the Catalytic Subunit of Protein Kinase 2 and Carries Histone-Like Lysine-Methylated Mark |
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Authors: | Ksenia S. Egorova Ludmila V. Olenina |
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Affiliation: | Laboratory of Biochemical Genetics of Animals, Institute of Molecular Genetics, Russian Academy of Science, 123182, Kurchatov Square, 2, Moscow, Russia |
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Abstract: | SummaryThe X-chromosome-linked clusters of the tandemly repeated testis-specific Stellate genes of Drosophila melanogaster, encoding proteins homologous to the regulatory β-subunit of the protein kinase casein kinase 2 (CK2), are repressed in wild-type males. Derepression of Stellate genes in the absence of the Y chromosome or Y-linked crystal locus (crystal line) causes accumulation of abundant protein crystals in testes and different meiotic abnormalities, which lead to partial or complete male sterility. To understand the cause of abnormalities in chromosome behavior owing to Stellate overexpression, we studied subcellular localization of Stellate proteins by biochemical fractionation and immunostaining of whole testes. We showed that, apart from the known accumulation of Stellate in crystalline form, soluble Stellate was located exclusively in the nucleoplasm, whereas Stellate crystals were located mainly in the cytoplasm. Coimmunoprecipitation experiments revealed that the α-subunit of the protein kinase CK2 (CK2α) was associated with soluble Stellate. Interaction between soluble Stellate and CK2α in the nucleus could lead to modulations in the phosphorylation of nuclear targets of CK2 and abnormalities in the meiotic segregation of chromosomes. We also observed that Stellate underwent lysine methylation and mimicked trimethyl-H3K9 epigenetic modification of histone H3 tail. |
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Keywords: | CK2, casein kinase 2 Su(Ste), suppressor of Stellate DAPI, 4&prime ,6-diamidino-2-phenylindole HKMT, histone lysine (K) methyltransferase PBS, phosphate-buffered saline EDTA, ethylenediaminetetraacetic acid EGTA, ethylene glycol bis(b-aminoethyl ether) N,N&prime -tetraacetic acid MALDI, matrix-assisted laser desorption/ionization |
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