title: Upregulation of Polo-like kinase 2 gene expression by GATA-1 acetylation in human osteosarcoma MG-63 cells.
abstract: Polo-like kinase 2 (Plk2) is a member of the serine/threonine protein kinase family involved in cell-cycle regulation and cellular response to stresses. It is of great interest to investigate the molecular mechanisms that control the expression of Plk2. Here, using real-time PCR and Western blot assays, we show that trichostatin A (TSA), a histone deacetylase inhibitor, upregulated Plk2 mRNA and protein expression in the human osteosarcoma MG-63 cell line. Luciferase activity analysis of the truncated Plk2 promoter indicated that the region from -1220 to -830 of the Plk2 promoter was sensitive to TSA. Moreover, using the electrophoresis mobility shift assay and chromatin immunoprecipitation assay, we identified two GATA-1 responsive elements at positions -1051 and -949, to which GATA-1 binding was enhanced by TSA under in vitro and in vivo conditions. Immunoprecipitation and Western blot showed that the levels of acetylated GATA-1 were increased with TSA in MG-63 cells, consistent with their binding affinities to the GATA-1 responsive elements. In summary, these data demonstrate that acetylation plays a crucial role in Plk2 expression and acetylation of GATA-1 by TSA treatment may upregulate their DNA-binding affinities, resulting in the activation of Plk2 promoter. These results may contribute to the understanding of the molecular mechanism of Plk2 regulation.
authors.affiliation: Department of Orthopedic Surgery, Shengjing Hospital, China Medical University, No. 36, Sanhao Street, Heping District, Shenyang 110004, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Shen
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Liqing
authors.initials: L
authors.lastname: Yang
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Xu
authors.forename: Feng
authors.initials: F
authors.lastname: Liang
authors.forename: Shuang
authors.initials: S
authors.lastname: Liang
authors.forename: Gen
authors.initials: G
authors.lastname: Ba
authors.forename: Feng
authors.initials: F
authors.lastname: Xue
authors.forename: Qin
authors.initials: Q
authors.lastname: Fu
mesh_terms: D000107:Acetylation / Q000187:drug effects; D045744:Cell Line, Tumor; D050982:GATA1 Transcription Factor / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D006877:Hydroxamic Acids / Q000008:administration & dosage / Q000494:pharmacology; D012516:Osteosarcoma; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; D006877:Hydroxamic Acids; C012589:trichostatin A; C496126:PLK2 protein, human; D017346:Protein Serine-Threonine Kinases