Negative Regulation of p21Waf1/Cip1 by Human INO80 Chromatin Remodeling Complex Is Implicated in Cell Cycle Phase G2/M Arrest and Abnormal Chromosome Stability

Lingling Cao; Jian Ding; Liguo Dong; Jiayao Zhao; Jiaming Su; Lingyao Wang; Yi Sui; Tong Zhao; Fei Wang; Jingji Jin; Yong Cai

doi:10.1371/journal.pone.0137411

Negative Regulation of p21Waf1/Cip1 by Human INO80 Chromatin Remodeling Complex Is Implicated in Cell Cycle Phase G2/M Arrest and Abnormal Chromosome Stability

PLoS One. 2015 Sep 4;10(9):e0137411. doi: 10.1371/journal.pone.0137411. eCollection 2015.

Authors

Lingling Cao¹, Jian Ding¹, Liguo Dong¹, Jiayao Zhao¹, Jiaming Su¹, Lingyao Wang¹, Yi Sui¹, Tong Zhao¹, Fei Wang¹, Jingji Jin², Yong Cai²

Affiliations

¹ School of Life Sciences, Jilin University, Changchun, Jilin, China.
² School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun City, Jilin, China; Key Laboratory for Molecular Enzymology and Engineering, the Ministry of Education, Jilin University, Changchun City, Jilin, China.

Abstract

We previously identified an ATP-dependent human Ino80 (INO80) chromatin remodeling complex which shares a set of core subunits with yeast Ino80 complex. Although research evidence has suggested that INO80 complex functions in gene transcription and genome stability, the precise mechanism remains unclear. Herein, based on gene expression profiles from the INO80 complex-knockdown in HeLa cells, we first demonstrate that INO80 complex negatively regulates the p21Waf1/Cip1 (p21) expression in a p53-mediated mechanism. In chromatin immunoprecipitation (ChIP) and a sequential ChIP (Re-ChIP) assays, we determined that the INO80 complex and p53 can bind to the same promoter region of p21 gene (-2.2 kb and -1.0 kb upstream of the p21 promoter region), and p53 is required for the recruitment of the INO80 complex to the p21 promoter. RNAi knockdown strategies of INO80 not only led to prolonged progression of cell cycle phase G2/M to G1, but it also resulted in abnormal chromosome stability. Interestingly, high expression of p21 was observed in most morphologically-changed cells, suggesting that negative regulation of p21 by INO80 complex might be implicated in maintaining the cell cycle process and chromosome stability. Together, our findings will provide a theoretical basis to further elucidate the cellular mechanisms of the INO80 complex.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATPases Associated with Diverse Cellular Activities
Binding Sites
Chromatin / chemistry
Chromatin / metabolism
Chromatin Assembly and Disassembly*
Chromatin Immunoprecipitation
Chromosomal Instability
Cyclin-Dependent Kinase Inhibitor p21 / genetics*
Cyclin-Dependent Kinase Inhibitor p21 / metabolism
DNA Helicases / antagonists & inhibitors
DNA Helicases / genetics*
DNA Helicases / metabolism
DNA-Binding Proteins
G2 Phase Cell Cycle Checkpoints / genetics*
Gene Expression Regulation
HCT116 Cells
HEK293 Cells
HeLa Cells
Humans
Oligonucleotide Array Sequence Analysis
Promoter Regions, Genetic
Protein Binding
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Signal Transduction
Tumor Suppressor Protein p53 / genetics*
Tumor Suppressor Protein p53 / metabolism

Substances

Chromatin
Cyclin-Dependent Kinase Inhibitor p21
DNA-Binding Proteins
RNA, Small Interfering
Tumor Suppressor Protein p53
ATPases Associated with Diverse Cellular Activities
DNA Helicases
INO80 protein, human

Grants and funding

This work was supported by National Natural Science Foundation of China (No. 31071131&31171245, YC), National Laboratory of Biomacromolecules, Institue of Biophysics, Chinese Academy of Sciences (No. O5SY02110A, YC and 2012kf04, JJ) and the Project of Jilin Province Science and Technology Development Program (20130413002GH, YC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.