CLONING AND CHARACTERIZATION OF A CDNA-ENCODING THE BETA-SUBUNIT OF HUMAN CASEIN KINASE-II
CLONING AND CHARACTERIZATION OF A CDNA-ENCODING THE BETA-SUBUNIT OF HUMAN CASEIN KINASE-II
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DOI：
10.1021/bi00449a014
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发表时间：
1989-11-14
期刊：
影响因子：
2.9
通讯作者：
CZECH, MP
中科院分区：
文献类型：
--
作者：
HELLERHARRISON, RA;MEISNER, H;CZECH, MP
先前的研究[Summercorn 等人。 (1987) 程序。国家。阿卡德。科学。美国 84, 8834-8838； Klarlund 和 Czech (1988) J. Biol。化学。 [263, 15872-15875] 表明，用胰岛素孵育的 Balb/c 3T3 细胞和 3T3-L1 脂肪细胞在几分钟内显示酪蛋白激酶 II 活性增加，表明这种丝氨酸/苏氨酸激酶是胰岛素信号传导途径的早期步骤。我们最近报道了编码 .alpha 的 cDNA 的分离。人酪蛋白激酶 II 亚基 [Meisner 等人。 (1989) Biochemistry 28, 4072-4076] 作为检查这种酶调节的第一步。我们现在描述 HepG2 细胞酪蛋白激酶 II .beta。 2.57 kb 的亚基 cDNA，包含 96 个碱基的 5'' 非翻译序列、645 个碱基的开放阅读框和 1832 个碱基的 3'' 非翻译序列，具有两个聚腺苷酸化共有信号序列和两个聚 (A) 序列。人类 .beta 的开放阅读框。亚基 cDNA 在核苷酸和氨基酸水平上分别与果蝇序列有 77% 和 87% 的同一性，与牛氨基酸序列有 99% 的同一性。利用 HepG2 .beta 对 HepG2 细胞 RNA 进行 RNA 分析。亚基 cDNA 片段作为探针，显示一条主要条带在 1.2 kb 处迁移，两条次要条带在 3.0 和 4.2 kb 处迁移。 HepG2 基因组 DNA 的 DNA 分析结果与利用果蝇基因组 DNA 的结果一致，表明存在 .beta 的单个基因。酪蛋白激酶 II 的亚基。
Previous studies [Summercorn et al. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 8834-8838; Klarlund and Czech (1988) J. Biol. Chem. 263, 15872-15875] have indicated that Balb/c 3T3 cells and 3T3-L1 adipocytes incubated with insulin show increased casein kinase II activity within minutes, implicating this serine/threonine kinase as an early step in an insulin signaling pathway. we recently reported the isolation of a cDNA encoding an .alpha. subunit of human casein kinase II [Meisner et al. (1989) Biochemistry 28, 4072-4076] as an initial step toward examining the regulation of this enzyme. We now describe a HepG2 cell casein kinase II .beta. subunit cDNA of 2.57 kb containing 96 bases of 5'' untranslated sequence, 645 bases of open reading frame, and 1832 bases of 3'' untranslated sequence with two polyadenylation consensus signal sequences and two poly(A) stretches. The open reading frame of the human .beta. subunit cDNA was 77% and 87% identical with the Drosophila sequence at the nucleotide and amino acid levels, respectively, and 99% identical with the bovine amino acid sequence. RNA analysis of HepG2 cell RNA utilizing HepG2 .beta. subunit cDNA fragments as probes revealed one major band migrating at 1.2 kb and two minor bands migrating at 3.0 and 4.2 kb. Results from DNA analysis of HepG2 genomic DNA, consistent with results utilizing Drosophila genomic DNA, suggest the presence of a single gene for the .beta. subunit of casein kinase II.