Negative Regulation of p21Waf1/Cip1 by Human INO80 Chromatin Remodeling Complex Is Implicated in Cell Cycle Phase G2/M Arrest and Abnormal Chromosome Stability.
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.
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Lingling
L
Cao
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Jian
J
Ding
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Liguo
L
Dong
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Jiayao
J
Zhao
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Jiaming
J
Su
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Lingyao
L
Wang
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Yi
Y
Sui
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Tong
T
Zhao
School of Life Sciences, Jilin University, Changchun, Jilin, China.
Fei
F
Wang
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.
Jingji
J
Jin
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.
Yong
Y
Cai
D000074183:ATPases Associated with Diverse Cellular Activities; D001665:Binding Sites; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation; D043171:Chromosomal Instability; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics* / Q000378:metabolism; D004265:DNA Helicases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins; D059565:G2 Phase Cell Cycle Checkpoints / Q000235:genetics*; D005786:Gene Expression Regulation; D045325:HCT116 Cells; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism
D002843:Chromatin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D004268:DNA-Binding Proteins; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D000074183:ATPases Associated with Diverse Cellular Activities; D004265:DNA Helicases; C524421:INO80 protein, human