cDNA cloning and chromosomal mapping of genes encoding novel protein kinases termed PKU-α and PKU-β, which have nuclear localization signal
cDNA cloning and chromosomal mapping of genes encoding novel protein kinases termed PKU-α and PKU-β, which have nuclear localization signal
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DOI：
10.1016/s0378-1119(97)00495-2
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发表时间：
1997-11-20
期刊：
影响因子：
3.5
通讯作者：
Date, T
中科院分区：
文献类型：
--
作者：
Yamakawa, A;Kameoka, Y;Date, T
我们已经克隆了新的丝氨酸/苏氨酸蛋白激酶（PK）的cDNA，称为PKU-α和PKU-β，通过筛选噬菌体表达库的激酶活性。对PKU-α和PKU-β基因的序列分析表明，它们的开放阅读框（ORF）分别为2151和2361个核苷酸（nt），分别编码727和787个氨基酸残基的多肽。PKU-α和PKU-β的氨基酸序列在C-末端区域含有典型的丝氨酸/苏氨酸PK结构域，同源性为86%，表明它们属于同一PK家族。北方分析表明，它们在几乎所有的人体组织和培养细胞中表达。通过荧光原位杂交将PKU-α和PKU-β的基因分别定位于染色体17 q23和8 p12-p22。这两种cDNA编码的蛋白质在其N-末端区域含有推定的核定位信号（NLS）。这些信号可能在核定位中起作用。谷氨酰胺S-转移酶（GST）-融合的区域PKU-α和β含有NLS有效地定位于细胞核。此外，在COS-1细胞中瞬时表达的PKU-β主要是核。PKU-α和PKU-β不同：在PKU-β的NLS附近存在nt结合基序的共有序列。这些结果表明，PKU-α和β可以磷酸化类似蛋白质上的丝氨酸和/或苏氨酸残基，但它们的活性通过与核组分的不同相互作用来调节。(C)1997年Elsevier Science B.V.
We have cloned cDNAs for novel serine/threonine protein kinases (PK), termed PKU-alpha and PKU-beta, by screening a bacteriophage expression library for kinase activity. Sequence analysis of PKU-alpha and PKU-beta genes revealed that their open reading frames (ORF) were 2151 and 2361 nucleotides (nt) encoding polypeptides of 727 and 787 amino acid (aa) residues, respectively. The deduced aa sequences of PKU-alpha and PKU-beta contained typical serine/threonine PK domains at the C-terminal region and were 86% identical to each other, indicating that they belong to the same PK family. Northern analysis reveals that they are expressed in nearly all human tissues and in cultured cells. The genes for PKU-alpha and PKU-beta were mapped to chromosome 17q23 and 8p12-p22, respectively, by fluorescence in situ hybridization. The proteins encoded by both cDNAs contain a putative nuclear localization signal (NLS) in their N-terminal region. These signals are likely to function in nuclear localization. Glutathione S-transferase (GST)-fusions to regions of PKU-alpha and beta containing the NLS were efficiently localized to the nucleus. In addition, PKU-beta transiently expressed in COS-1 cells was predominantly nuclear. PKU-alpha and PKU-beta differ: a consensus sequence for a nt binding motif is present near the NLS of PKU-beta. These results suggest that PKU-alpha and beta may phosphorylate serine and/or threonine residues on similar proteins, but their activities are regulated through distinct interactions with a nuclear component. (C) 1997 Elsevier Science B.V.