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.

[1]  J. Swedlow,et al.  Aurora B regulates MCAK at the mitotic centromere. , 2004, Developmental cell.

[2]  J. Shabanowitz,et al.  Aurora B Phosphorylates Centromeric MCAK and Regulates Its Localization and Microtubule Depolymerization Activity , 2004, Current Biology.

[3]  D. Compton,et al.  The kinesin-13 proteins Kif2a, Kif2b, and Kif2c/MCAK have distinct roles during mitosis in human cells. , 2007, Molecular biology of the cell.

[4]  N. Hirokawa,et al.  Kinesin superfamily proteins and their various functions and dynamics. , 2004, Experimental cell research.

[5]  C. Walczak,et al.  The microtubule-destabilizing kinesin XKCM1 regulates microtubule dynamic instability in cells. , 2002, Molecular biology of the cell.

[6]  Brian David Dynlacht,et al.  E2F mediates cell cycle-dependent transcriptional repression in vivo by recruitment of an HDAC1/mSin3B corepressor complex. , 2002, Genes & development.

[7]  Y. H. Kim,et al.  Cloning and expression of human mitotic centromere-associated kinesin gene. , 1997, Biochimica et biophysica acta.

[8]  E. E. Brooks,et al.  Functional Analysis of the Human Cyclin D2 and Cyclin D3 Promoters (*) , 1996, The Journal of Biological Chemistry.

[9]  E. Ziff,et al.  Nerve Growth Factor Induces Transcription of the p21 WAF1/CIP1 and Cyclin D1 Genes in PC12 Cells by Activating the Sp1 Transcription Factor , 1997, The Journal of Neuroscience.

[10]  M. Wagenbach,et al.  MUTATIONS IN THE ATP‐BINDING DOMAIN AFFECT THE SUBCELLULAR DISTRIBUTION OF MITOTIC CENTROMERE‐ASSOCIATED KINESIN (MCAK) , 1999, Cell biology international.

[11]  K. Hunt,et al.  E2F–HDAC complexes negatively regulate the tumor suppressor gene ARHI in breast cancer , 2006, Oncogene.

[12]  K. Mimori,et al.  Clinicopathological and biological significance of mitotic centromere-associated kinesin overexpression in human gastric cancer , 2007, British Journal of Cancer.

[13]  H. Niwa,et al.  Efficient selection for high-expression transfectants with a novel eukaryotic vector. , 1991, Gene.

[14]  S. Ems-McClung,et al.  The interplay of the N- and C-terminal domains of MCAK control microtubule depolymerization activity and spindle assembly. , 2006, Molecular biology of the cell.

[15]  Yamamura Ken-ichi,et al.  Efficient selection for high-expression transfectants with a novel eukaryotic vector , 1991 .

[16]  Jonathon Howard,et al.  The depolymerizing kinesin MCAK uses lattice diffusion to rapidly target microtubule ends , 2006, Nature.

[17]  J. Azizkhan,et al.  Transcriptional initiation is controlled by upstream GC-box interactions in a TATAA-less promoter , 1990, Molecular and cellular biology.

[18]  Russell L. Malmberg,et al.  A standardized kinesin nomenclature , 2004, The Journal of cell biology.

[19]  L. Lania,et al.  Transcriptional regulation by the Sp family proteins. , 1997, The international journal of biochemistry & cell biology.

[20]  K. Helin,et al.  The E2F family: specific functions and overlapping interests , 2004, The EMBO journal.

[21]  G. Suske.  The Sp-family of transcription factors. , 1999, Gene.

[22]  D. Compton,et al.  The KinI kinesin Kif2a is required for bipolar spindle assembly through a functional relationship with MCAK , 2004, The Journal of cell biology.

[23]  T. Mitchison,et al.  Identification and partial characterization of mitotic centromere- associated kinesin, a kinesin-related protein that associates with centromeres during mitosis , 1995, The Journal of cell biology.

[24]  A. Blais,et al.  Hitting their targets: an emerging picture of E2F and cell cycle control. , 2004, Current opinion in genetics & development.