CLONING OF A HUMAN INSULIN-STIMULATED PROTEIN-KINASE (ISPK-1) GENE AND ANALYSIS OF CODING REGIONS AND MESSENGER-RNA LEVELS OF THE ISPK-1 AND THE PROTEIN PHOSPHATASE-1 GENES IN MUSCLE FROM NIDDM PATIENTS
CLONING OF A HUMAN INSULIN-STIMULATED PROTEIN-KINASE (ISPK-1) GENE AND ANALYSIS OF CODING REGIONS AND MESSENGER-RNA LEVELS OF THE ISPK-1 AND THE PROTEIN PHOSPHATASE-1 GENES IN MUSCLE FROM NIDDM PATIENTS
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DOI：
10.2337/diabetes.44.1.90
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发表时间：
1995-01-01
期刊：
影响因子：
7.7
通讯作者：
PEDERSEN, O
中科院分区：
文献类型：
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作者：
BJORBAEK, C;VIK, TA;PEDERSEN, O
对编码蛋白磷酸酶 1（PP1 α、PP1 β 和 PP1 γ）和胰岛素刺激蛋白激酶 1 (ISPK-1) 的三个催化亚基和胰岛素刺激蛋白激酶 1 (ISPK-1) 的互补 DNA 进行分析，以了解与 30 名非胰岛素依赖型糖尿病患者骨骼肌中胰岛素抵抗糖原合成相关的编码区的变化 梅利图斯（NIDDM）。人类 ISPK-1 cDNA 克隆至人类 T 细胞白血病和胎盘 cDNA 文库，并定位到人类 X 染色体的短臂。单链构象多态性 (SSCP) 分析确定了 PP1 基因编码区总共有 6 个变异：其中两个位于 PP1 α 中。密码子 90 和 255； PP1 beta 密码子 67 处有一个； PP1 gamma 中的 3 个分别位于密码子 11、269 和 273 处。然而，所有这些都只是沉默的单核苷酸取代。 ISPK-1 基因的 SSCP 分析确定了密码子 266 处的一个沉默多态性和密码子 38 处的一个氨基酸变异（Ile-->Ser）。该变异主要发现于一名男性 NIDDM 患者。然而，该受试者并未表现出肌肉胰岛素刺激的糖原合酶激活受损。在 15 名 NIDDM 患者和 14 名健康受试者之间，肌肉中这四种基因的 mRNA 水平没有发现显着差异。我们的研究结果表明：1) PP1 α、PP1 β、PP1 γ 和 ISPK-1 编码区的遗传异常不太可能是 NIDDM 患者分析组肌肉中胰岛素刺激的糖原合成激活减少的常见原因； 2) NIDDM患者肌角PP1α、PP1β、PP1γ、ISPK-1 mRNA水平正常； 3) 胰岛素抵抗 NIDDM 患者中胰岛素刺激肌糖原合成激活的推定遗传缺陷可能位于胰岛素作用级联中 ISPK-1 的更上游。
Complementary DNA encoding three catalytic subunits of protein phosphatase 1 (PP1 alpha, PP1 beta, and PP1 gamma) and the insulin-stimulated protein kinase 1 (ISPK-1) was analyzed for variations in the coding regions related to insulin-resistant glycogen synthesis in skeletal muscle of 30 patients with non-insulin-dependent diabetes mellitus (NIDDM). The human ISPK-1 cDNA was cloned hom T-cell leukemia and placental cDNA libraries and mapped to the short arm of the human X chromosome. Single-strand conformation polymorphism (SSCP) analysis identified a total of six variations in the coding regions of the PP1 genes: two in PP1 alpha. at codons 90 and 255; one in PP1 beta at codon 67; and three in PP1 gamma at codons 11, 269, and 273, respectively. All mere, however, silent single nucleotide substitutions. SSCP analysis of the ISPK-1 gene identified one silent polymorphism at codon 266 and one amino acid variant at codon 38 (Ile-->Ser). This variant was primarily found in one male NIDDM patient. This subject, however, did not exhibit an impairment of muscle insulin-stimulated glycogen synthase activation. No significant differences were found in mRNA levels in muscle of the four genes between 15 NIDDM patients and 14 healthy subjects. Our findings suggest that 1) genetic abnormalities in the coding regions of PP1 alpha, PP1 beta, PP1 gamma, and ISPK-1 are unlikely to be frequently occurring causes of the reduced insulin-stimulated activation of the glycogen synthesis in muscle from the analyzed group of NIDDM patients; 2) the mRNA levels of PP1 alpha, PP1 beta, PP1 gamma, and ISPK-1 are normal in muscle horn the NIDDM patients; and 3) putative inherited defects in insulin-stimulated activation of muscle glycogen synthesis in patients with insulin-resistant NIDDM may be located further upstream of ISPK-1 in the insulin action cascade.