Conformational Change of Human Checkpoint Kinase 1 (Chk1) Induced by DNA Damage
| Autor: | Feng Ren, Xin-Sheng Yao, Krzysztof Palczewski, Jingna Wang, Philip D. Kiser, Xiangzi Han, Paul S.-H. Park, Jinshan Tang, Jinhua Zheng, Megan Gragg, Youwei Zhang |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Conformational change animal structures Protein Conformation DNA damage genetic processes DNA and Chromosomes Biology medicine.disease_cause environment and public health Biochemistry Cell Line 03 medical and health sciences Protein structure Fluorescence Resonance Energy Transfer medicine Humans Protein Interaction Domains and Motifs CHEK1 Phosphorylation Molecular Biology Genetics Mutation Cell Biology enzymes and coenzymes (carbohydrates) HEK293 Cells 030104 developmental biology Förster resonance energy transfer Protein kinase domain Checkpoint Kinase 1 Biophysics biological phenomena cell phenomena and immunity Protein Kinases DNA Damage |
| Zdroj: | Journal of Biological Chemistry. 291:12951-12959 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m115.713248 |
| Popis: | Phosphorylation of Chk1 by ataxia telangiectasia-mutated and Rad3-related (ATR) is critical for checkpoint activation upon DNA damage. However, how phosphorylation activates Chk1 remains unclear. Many studies suggest a conformational change model of Chk1 activation in which phosphorylation shifts Chk1 from a closed inactive conformation to an open active conformation during the DNA damage response. However, no structural study has been reported to support this Chk1 activation model. Here we used FRET and bimolecular fluorescence complementary techniques to show that Chk1 indeed maintains a closed conformation in the absence of DNA damage through an intramolecular interaction between a region (residues 31–87) at the N-terminal kinase domain and the distal C terminus. A highly conserved Leu-449 at the C terminus is important for this intramolecular interaction. We further showed that abolishing the intramolecular interaction by a Leu-449 to Arg mutation or inducing ATR-dependent Chk1 phosphorylation by DNA damage disrupts the closed conformation, leading to an open and activated conformation of Chk1. These data provide significant insight into the mechanisms of Chk1 activation during the DNA damage response. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|