Cell-cycle-dependent regulation of human aurora A transcription is mediated by periodic repression of E4TF1.
Human aurora A is a serine-threonine kinase that controls various mitotic events. The transcription of aurora A mRNA varies throughout the cell cycle and peaks during G(2)/M. To clarify the transcriptional mechanism, we first cloned the 1.8-kb 5'-flanking region of aurora A including the first exon. Transient expression of aurora A promoter-luciferase constructs containing a series of 5'-truncated sequences or site-directed mutations identified a 7-bp sequence (CTTCCGG) from -85 to -79 as a positive regulatory element. Electromobility shift assays identified the binding of positive regulatory proteins to the CTTCCGG element. Anti-E4TF1-60 antibody generated a supershifted complex. Furthermore, coexpression of E4TF1-60 and E4TF1-53 markedly increased aurora A promoter activity. Synchronized cells transfected with the aurora A promoter-luciferase constructs revealed that the promoter activity of aurora A increased in the S phase and peaked at G(2)/M. In addition, we identified a tandem repressor element, CDE/CHR, just downstream of the CTTCCGG element, and mutation within this element led to a loss of cell cycle regulation. We conclude that the transcription of aurora A is positively regulated by E4TF1, a ubiquitously expressed ETS family protein, and that the CDE/CHR element was essential for the G(2)/M-specific transcription of aurora A.
Tanaka
Masatsugu
M
First Department of Internal Medicine, Yokohama City University School of Medicine, Yokohama 236-0004, Japan.
Ueda
Atsuhisa
A
Kanamori
Heiwa
H
Ideguchi
Haruko
H
Yang
Jun
J
Kitajima
Shigetaka
S
Ishigatsubo
Yoshiaki
Y
D016475:3T3 Cells; D000818:Animals; D064096:Aurora Kinase A; D064127:Aurora Kinases; D001483:Base Sequence; D015152:Blotting, Northern; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D005091:Exons; D051268:GA-Binding Protein Transcription Factor; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D007834:Lasers; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011494:Protein Kinases / Q000096:biosynthesis* / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D029867:Xenopus Proteins
D018797:Cell Cycle Proteins; C079860:E4TF1 transcription factor; D051268:GA-Binding Protein Transcription Factor; D014157:Transcription Factors; D029867:Xenopus Proteins; D008156:Luciferases; D011494:Protein Kinases; C112134:AURKA protein, Xenopus; C576918:Aurka protein, mouse; D064096:Aurora Kinase A; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases