PU.1 can regulate the ZNF300 promoter in APL-derived promyelocytes HL-60.
ZNF300, which plays the role in human embryonic development and some diseases, is a typical KRAB/C2H2 zinc finger gene expressed only in higher mammalians. Our data showed that expression of ZNF300 changed significantly in various leukemia blasts in the bone marrow aspirates of newly diagnosed leukemia patients. To investigate the potential relationship between expression of ZNF300 and the progression of leukemia development and hematopoietic differentiation, we cloned and characterized the putative human ZNF300 gene promoter and identified its transcription start sites (TSSs). Deletion and mutagenesis analysis demonstrated that a myeloid-specific transcription factor PU.1 binding site was responsible for myeloid-specific regulation of ZNF300 promoter activity. Furthermore, electrophoretic mobility shift and chromatin immunoprecipitation assays revealed that PU.1 bound to the PU.1 binding site within ZNF300 promoter region in vitro and in vivo. Overexpression of PU.1 elevated ZNF300 promoter activity, whereas silencing of PU.1 expression significantly reduced the activity in myeloid-derived HL-60 cell but not in T-cell Jurkat. In vitro induced HL-60 cells into CD11b expressing cells by DMSO demonstrated that ZNF300 was upregulated along with upregulation of PU.1 expression. These results demonstrated that ZNF300 was activated by PU.1 and suggested that the regulation may be involved in the progression of leukemia development and hematopoietic differentiation.
Xu
Jun-Hua
JH
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Luojia Hill, Wuchang, Wuhan, PR China.
Wang
Tao
T
Wang
Xian-Guo
XG
Wu
Xiang-Peng
XP
Zhao
Zhou-Zhou
ZZ
Zhu
Chen-Gang
CG
Qiu
Hong-Ling
HL
Xue
Lu
L
Shao
Huan-Jie
HJ
Guo
Ming-Xiong
MX
Li
Wen-Xin
WX
D039481:CD11b Antigen / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D015973:Gene Expression Regulation, Leukemic; D018922:HL-60 Cells; D006801:Humans; D019169:Jurkat Cells; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D020218:Response Elements; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D000099232:Proto-Oncogene Protein Spi-1
D039481:CD11b Antigen; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1; C509295:ITGAM protein, human; C485905:ZNF300 protein, human