Identification of kinases phosphorylating 13 sites in the nuclear, DNA-binding protein NUCKS

Grundt Kirsten; Thiede Bernd; Ostvold Anne Carine

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Identification of kinases phosphorylating 13 sites in the nuclear, DNA-binding protein NUCKS

机译：鉴定激酶磷酸化13位核，DNA结合蛋白咬伤

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NUCKS is a vertebrate specific, nuclear and DNA-binding phospho protein. The protein is highly expressed in rapidly dividing cells, and is overexpressed in a number of cancer tissues. The phosphorylation of NUCKS is cell cycle and DNA-damage regulated, but little is known about the responsible kinases. By utilizing in vitro and in vivo phosphorylation assays using isolated NUCKS as well as synthetic NUCKS-derived peptides in combination with mass spectrometry, phosphopeptide mapping, phosphphoamino acid analyses, phosphospecific antibodies and the use of specific kinase inhibitors, we found that NUCKS is phosphorylated on 11 sites by CK2. At least 7 of the CK2 sites are phosphorylated in vivo. We also found that NUCKS is phosphorylated on two sites by ATM kinase and DNA-PK in vitro, and is phosphorylated in vivo by ATM kinase in gamma-irradiated cells. All together, we identified three kinases phosphorylating 13 out of 39 in vivo phosphorylated sites in mammalian NUCKS. The identification of CK2 and PIKK kinases as kinases phosphorylating NUCKS in vivo provide further evidence for the involvement of NUCKS in cell cycle control and DNA repair. (C) 2017 Elsevier B.V. All rights reserved.

机译：尼克斯是一种脊椎动物特异性，核和DNA结合磷蛋白。蛋白质在快速分开的细胞中高度表达，并且在许多癌组织中过表达。咬伤的磷酸化是细胞周期和调节的DNA损伤，但对负责的激酶有几乎是已知的。通过在体外和体内磷酸化测定中使用分离的咬合以及合成咬合捕获的肽，以及与质谱，磷肽映射，磷酸氨基酸分析，磷酸化抗体和使用特异性激酶抑制剂的使用，我们发现咬伤是磷酸化的CK2的11个网站。至少7个CK2位点在体内磷酸化。我们还发现，在体外通过ATM激酶和DNA-PK在两个位点上磷酸化，并且在γ-辐照细胞中通过ATM激酶体内磷酸化。一致，我们鉴定了三种激酶在哺乳动物尼克斯的体内磷酸化位点中的39位磷酸化13中。 CK2和PIKK激酶的鉴定为激进酶磷酸化咬合在体内捕获的咬合提供了进一步的禁止咬伤在细胞周期控制和DNA修复中的依据。（c）2017 Elsevier B.v.保留所有权利。

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《Biochimica et biophysica acta: BBA: International journal of biochemistry, biophysics and molecular biololgy. Proteins and Proteomics》 |2017年第3期|共11页
作者
Grundt Kirsten; Thiede Bernd; Ostvold Anne Carine;
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作者单位

Univ Oslo Inst Basic Med Sci Dept Biochem POB 1112 N-0317 Oslo Norway;

Univ Oslo Dept Biosci POB 1066 N-0316 Oslo Norway;

Univ Oslo Inst Basic Med Sci Dept Biochem POB 1112 N-0317 Oslo Norway;

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正文语种 eng
中图分类生物化学;
关键词
Phosphorylation; Cell cycle; CK2; ATM kinase; DNA-PK; DNA repair;

机译：磷酸化;细胞周期;CK2;ATM激酶;DNA-PK;DNA修复;

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专利

1. Identification of kinases phosphorylating 13 sites in the nuclear, DNA-binding protein NUCKS [J] . Grundt Kirsten, Thiede Bernd, Ostvold Anne Carine Biochimica et biophysica acta: BBA: International journal of biochemistry, biophysics and molecular biololgy. Proteins and Proteomics . 2017,第3期

机译：鉴定激酶磷酸化13位核，DNA结合蛋白咬伤
2. Identification and characterization of two putative nuclear localization signals (NLS) in the DNA-binding protein NUCKS. [J] . Grundt K, Haga IV, Huitfeldt HS, Biochimica et biophysica acta. Biomembranes . 2007,第9期

机译：DNA结合蛋白咬伤中的两个推定核定位信号（NLS）的鉴定和表征。
3. Identification of proteins directly phosphorylated by UL13 protein kinase from herpes simplex virus 1. [J] . Asai R, Ohno T, Kato A, Microbes and infection . 2007,第12a13期

机译：鉴定单纯疱疹病毒1中被UL13蛋白激酶直接磷酸化的蛋白。
4. First-time Identification of Four Protein Kinase A Phosphorylation Sites in Both Murine and Human Isoforms of Cardiac Myosin Binding Protein-C [C] . Weitao Jia, Justin F. Shaffer, Samantha P. Harris, American Society for Mass Spectrometry Conference on Mass Spectrometry and Allied Topics . 2009

机译：四种蛋白激酶的首次鉴定鼠和人同种型的磷酸化位点结合蛋白-c-c
5. The simian cytomegalovirus assembly protein precursor is phosphorylated at a glycogen synthase kinase-3 site and its downstream “priming” site: Evidence that phosphorylation affects its molecular interactions [D] . Casaday, Rebecca J. 2005

机译：猿猴巨细胞病毒装配蛋白前体在糖原合酶激酶-3位点及其下游“启动”位点被磷酸化：磷酸化影响其分子相互作用的证据
6. Herpes Simplex Virus 1-Encoded Protein Kinase UL13 Phosphorylates Viral Us3 Protein Kinase and Regulates Nuclear Localization of Viral Envelopment Factors UL34 and UL31 [O] . Akihisa Kato, Mayuko Yamamoto, Takashi Ohno, 2006

机译：单纯疱疹病毒1编码的蛋白激酶UL13磷酸化病毒us3蛋白激酶并调节病毒包膜因子UL34和UL31的核定位。
7. Identification and characterization of two putative nuclear localization signals (NLS) in the DNA-binding protein NUCKS [O] . Grundt Kirsten, Haga Ingvild V., Huitfeldt Henrik S., 2007

机译：DNA结合蛋白NUCKS中两个推定的核定位信号（NLS）的鉴定和表征
8. Characterization of the Sites Phosphorylated on Tyrosine Hydroxylase by Ca(2+) and Phospholipid-Dependent Protein Kinase, Calmodulin-Dependent Multiprotein Kinase and Cyclic AMP-Dependent Protein Kinase [R] . Vulliet, P. R., Woodgett, J. R., Ferrari, S., 1985

机译：通过Ca（2+）和磷脂依赖性蛋白激酶，钙调蛋白依赖性多蛋白激酶和环amp依赖性蛋白激酶在酪氨酸羟化酶上磷酸化位点的表征

Identification of kinases phosphorylating 13 sites in the nuclear, DNA-binding protein NUCKS

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