TISSUE-SPECIFIC AND CELL-SPECIFIC EXPRESSION OF INS(1,4,5)P-3 3-KINASE ISOENZYMES
TISSUE-SPECIFIC AND CELL-SPECIFIC EXPRESSION OF INS(1,4,5)P-3 3-KINASE ISOENZYMES
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DOI：
10.1042/bj3060429
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发表时间：
1995-03-01
影响因子：
4.1
通讯作者：
ERNEUX, C
中科院分区：
文献类型：
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作者：
VANWEYENBERG, V;COMMUNI, D;ERNEUX, C
InsP（3）3-激酶催化InsP（1，4，5）P-3（InsP（3））磷酸化为InsP（1，3，4，5）P-4（InsP（4））。分子生物学数据显示存在InsP（3）3-激酶的两种人类同工酶，即InsP（3）3-激酶A和B。我们从大鼠胸腺cDNA文库中分离到一个2235 bp的编码大鼠InsP（3）3-激酶B的cDNA（克隆B15）。对从大鼠组织（胸腺、睾丸、脑、脾、肝、肾、心脏、肺和肠）中分离的mRNA进行Northern印迹分析显示，大鼠InsP（3）3-激酶B探针与肺、胸腺、睾丸、脑和心脏中的6 kb mRNA杂交。相反，在严格条件下，用大鼠InsP（3）3-激酶A探针对相同组织进行Northern印迹分析，该探针仅与脑中的2kb mRNA杂交，与睾丸中的1.8- 2.0kb mRNA杂交。用人InsP（3）3-激酶B探针探测的三种人细胞系（HL-60、SH-SY 5 Y和HT B-138）的Northern印迹分析表明，除了在人神经母细胞瘤细胞系（SH-SY 5 Y）中检测到5.7和6 kb的两种mRNA外，所有细胞系中均存在6 kb的mRNA。使用相同的印迹，用人InsP（3）3-激酶A探针没有观察到杂交信号。总之，我们的数据与InsP（3）3-激酶同工酶A和B在不同组织和细胞中特异性表达的观点一致。
The phosphorylation of Ins(1,4,5)P-3 (InsP(3)) to Ins(1,3,4,5)P-4 (InsP(4)) is catalysed by InsP(3) 3-kinase. Molecular-biological data have shown the presence of two human isoenzymes of InsP(3) 3-kinase, namely InsP(3) 3-kinases A and B. We have isolated from a rat thymus cDNA library a 2235 bp cDNA (clone B15) encoding rat InsP(3) 3-kinase B. Northern-blot analysis of mRNA isolated from rat tissues (thymus, testis, brain, spleen, liver, kidney, heart, lung and intestine) revealed that a rat InsP(3) 3-kinase B probe hybridized to a 6 kb mRNA in lung, thymus, testis, brain and heart. In contrast, Northern-blot analysis of the same tissues probed under stringent conditions with a rat InsP(3) 3-kinase A probe hybridized to a 2 kb mRNA only in brain and a 1.8-2.0 kb mRNA species in testis. Northern-blot analysis of three human cell lines (HL-60, SH-SY5Y and HTB-138) probed with a human InsP(3) 3-kinase B probe showed the presence of a 6 kb mRNA in all cell lines, except in the human neuroblastoma cell line (SH-SY5Y), where two mRNA species of 5.7 and 6 kb were detected. Using the same blot, no hybridization signal could be seen with a human InsP(3) 3-kinase A probe. Altogether, our data are consistent with the notion that the two InsP(3) 3-kinase isoenzymes, A and B, are specifically expressed in different tissues and cells.