tailieunhanh - Báo cáo Y học: CK2btes gene encodes a testis-specific isoform of the regulatory subunit of casein kinase 2 in Drosophila melanogaster

An earlier described CK2btes gene of Drosophila melano-gasteris shown to encode amale germline speci®c isoformof regulatory bsubunit of casein kinase 2. Western-analysis using anti-CK2btes Ig revealed CK2btes protein in Drosophila testes extract. Expression of a CK2btes± b-galactosidase fusion protein driven by the CK2btes pro-moter was found in transgenic ¯ies at postmitotic stages of spermatogenesis. | Eur. J. Biochem. 269 1418-1427 2002 FEBS 2002 CK2ptes gene encodes a testis-specific isoform of the regulatory subunit of casein kinase 2 in Drosophila melanogaster Alla I. Kalmykova1 Yuri Y. Shevelyov1 Oksana O. Polesskaya1 Anna A. Dobritsa1 t Alexandra G. Evstafeva2 Brigitte Boldyreff3 Olaf-Georg Issinger3 and Vladimir A. Gvozdev1 Institute of Molecular Genetics Russian Academy of Sciences Moscow Russia 2Belozersky Institute of Physico-Chemical Biology Center of Molecular Medicine Moscow State University Russia 3Department of Biochemistry and Molecular Biology University of Southern Denmark Odense Denmark An earlier described CK2btes gene of Drosophila melano-gaster is shown to encode a male germline specific isoform of regulatory b subunit of casein kinase 2. Western-analysis using anti-CK2ptes Ig revealed CK2ptes protein in Drosophila testes extract. Expression of a CK2btes b-galactosidase fusion protein driven by the CK2btes promoter was found in transgenic flies at postmitotic stages of spermatogenesis. Examination of biochemical characteristics of a recombinant CK2btes protein expressed in Escherichia coli revealed properties similar to those of CK2b a CK2btes protein stimulates CK2a catalytic activity toward synthetic peptide b it inhibits phosphorylation of calmodulin and mediates stimulation of CK2a by polylysine c it is able to form CK2btes 2 dimers as well as CK2a 2 CK2btes 2 tetramers. Using the yeast two-hybrid system and coimmunoprecipitation analysis of protein extract from Drosophila testes we demonstrated an association between CK2btes and CK2a. Northern-analysis has shown that another regulatory b subunit found recently in D. melanogaster genome is also testis-specific. Thus we describe the first example of two tissue-specific regulatory subunits of CK2 which might serve to provide CK2 substrate recognition during spermatogenesis. Keywords spermatogenesis casein kinase 2 CK2 b subunit CK2btes testes. Protein kinase CK2 is involved in such general