Regulation of the human mitotic centromere-associated kinesin (MCAK) promoter by the transcription factors Sp1 and E2F1.
To understand transcriptional regulation of the human mitotic centromere-associated kinesin (MCAK) promoter, the 1,151-bp promoter region of the human MCAK gene in Jurkat T cells was cloned by polymerase chain reaction (PCR). Although a bioinformatic analysis of the promoter sequence predicted several putative transcription factor binding sites for E2F, Sp1, c-Myb, p53, p300, NF-1, AML-1a, Ap-1, E-box factor, and C/EBPalpha/beta with no consensus TATA-box motif, deletion constructs of the promoter region revealed that the core positive promoter activity resided at -266/-66, containing three GC-motifs for binding Sp1. Site-directed disruption and chromatin immunoprecipitation analysis indicated that Sp1-binding to the GC-motifs was crucial for promoter activation, but the E2F1-binding to the E2F-motif (-57/-50) was crucial for promoter repression. Cotransfection of the luciferase reporter with either Sp1- or E2F1-expression plasmid further verified the role of Sp1 as a transcriptional activator and E2F1 as a transcriptional repressor in the human MCAK promoter.
Jun
Do Youn
DY
Laboratory of Immunobiology, School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
Park
Hae-Sun
HS
Lee
Ji-Young
JY
Kim
Young Ho
YH
D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D050687:E2F1 Transcription Factor / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D019169:Jurkat Cells; D016547:Kinesins / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D015533:Transcriptional Activation
D050687:E2F1 Transcription Factor; C091489:KIF2C protein, human; D012097:Repressor Proteins; D016329:Sp1 Transcription Factor; D016547:Kinesins