Meisner H, Heller-Harrison R, Buxton J, Czech M P
Department of Biochemistry, University of Massachusetts Medical School, Worcester 01655.
Biochemistry. 1989 May 2;28(9):4072-6. doi: 10.1021/bi00435a066.
A human cDNA encoding the alpha subunit of casein kinase II and a partial cDNA encoding the rat homologue were isolated by using a Drosophila casein kinase II cDNA probe. The 2.2-kb human cDNA contains a 1.2-kb open reading frame, 150 nucleotides of 5' leader, and 850 nucleotides of 3' noncoding region. Except for the first 7 deduced amino acids that are missing in the rat cDNA, the 328 amino acids beginning with the amino terminus are identical between human and rat. The Drosophila enzyme sequence is 90% identical with the human casein kinase II sequence, and there is only a single amino acid difference between the published partial bovine sequence and the human sequence. In addition, the C-terminus of the human cDNA has an extra 53 amino acids not present in Drosophila. Northern analysis of rat and human RNA showed predominant bands of 5.5, 3.1, and 1.8 kb. In rat tissues, brain and spleen had the highest levels of casein kinase II alpha subunit specific RNA, while skeletal muscle showed the lowest. Southern analysis of human cultured cell and tissue genomic DNA using the full-length cDNA probe revealed two bands with restriction enzymes that have no recognition sites within the cDNA and three to six bands with enzymes having single internal sites. These results are consistent with the possibility that two genes encode the alpha subunits.
利用果蝇酪蛋白激酶II cDNA探针分离出了编码人酪蛋白激酶IIα亚基的cDNA以及编码大鼠同源物的部分cDNA。这个2.2kb的人cDNA包含一个1.2kb的开放阅读框、150个核苷酸的5'前导序列和850个核苷酸的3'非编码区。除了大鼠cDNA中缺失的前7个推导氨基酸外，从氨基末端开始的328个氨基酸在人和大鼠之间是相同的。果蝇酶序列与人类酪蛋白激酶II序列的同源性为90%，已发表的部分牛序列与人类序列之间只有一个氨基酸差异。此外，人cDNA的C末端还有额外的53个果蝇中不存在的氨基酸。对大鼠和人RNA的Northern分析显示，主要条带分别为5.5kb、3.1kb和1.8kb。在大鼠组织中，脑和脾中酪蛋白激酶IIα亚基特异性RNA水平最高，而骨骼肌中最低。用全长cDNA探针对人培养细胞和组织基因组DNA进行Southern分析，发现使用在cDNA内无识别位点的限制酶时出现两条带，使用有单个内部位点的酶时出现三到六条带。这些结果与两个基因编码α亚基的可能性一致。
