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一种保守的核丝氨酸（苏氨酸）蛋白激酶的分子克隆与特性分析

Molecular cloning and characterization of a conserved nuclear serine(threonine) protein kinase.

作者信息

Millward T, Cron P, Hemmings B A

机构信息

Friedrich Miescher-Institut, Basel, Switzerland.

出版信息

Proc Natl Acad Sci U S A. 1995 May 23;92(11):5022-6. doi: 10.1073/pnas.92.11.5022.

DOI:10.1073/pnas.92.11.5022
PMID:7761441
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC41840/
Abstract

Human, Drosophila melanogaster, and Caenorhabditis elegans cDNA clones encoding homologues of a serine(threonine) protein kinase (EC 2.7.1.37) (designated Ndr protein kinase) have been isolated and sequenced. The human and Drosophila cDNAs predict polypeptides of 54 kDa and 52 kDa, respectively, which share approximately 80% amino acid similarity. Northern analysis of human tissues revealed a ubiquitously expressed 3.9-kb transcript. Recombinant GST-Ndr underwent intramolecular autophosphorylation on serine and threonine residues in vitro but failed to transphosphorylate several standard protein kinase substrates. Transfection of the human cDNA into COS-1 cells resulted in the appearance of an intense nuclear staining in cells analyzed by indirect immunofluorescence; deletion mutagenesis identified a short basic peptide, KRKAETWKRNRR, responsible for the nuclear accumulation of Ndr. Thus, Ndr is a conserved and widely expressed nuclear protein kinase. The closest known relative of this previously uncharacterized kinase is Dbf2, a budding yeast protein kinase required for the completion of nuclear division.

摘要

已分离并测序了编码丝氨酸（苏氨酸）蛋白激酶（EC 2.7.1.37）（命名为Ndr蛋白激酶）同源物的人、黑腹果蝇和秀丽隐杆线虫的cDNA克隆。人及果蝇的cDNA分别预测出54 kDa和52 kDa的多肽，它们的氨基酸相似性约为80%。对人体组织进行的Northern分析显示有一个普遍表达的3.9 kb转录本。重组GST-Ndr在体外对丝氨酸和苏氨酸残基进行分子内自磷酸化，但未能对几种标准蛋白激酶底物进行转磷酸化。将人cDNA转染到COS-1细胞中，通过间接免疫荧光分析发现细胞中出现强烈的核染色；缺失诱变鉴定出一个短的碱性肽KRKAETWKRNRR，它负责Ndr的核积累。因此，Ndr是一种保守且广泛表达的核蛋白激酶。这种以前未被表征的激酶最接近的已知亲属是Dbf2，一种参与核分裂完成所需的芽殖酵母蛋白激酶。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/db2fa0b36728/pnas01487-0314-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/85b7c3eaaf10/pnas01487-0313-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/913e5bcc829a/pnas01487-0313-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/db2fa0b36728/pnas01487-0314-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/85b7c3eaaf10/pnas01487-0313-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/913e5bcc829a/pnas01487-0313-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b77c/41840/db2fa0b36728/pnas01487-0314-a.jpg

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