Zinc finger protein HZF1 promotes K562 cell proliferation by interacting with and inhibiting INCA1.

Previously, we characterized a zinc finger protein gene HZF1 (ZNF16) and demonstrated that it played a significant role in the erythroid and megakaryocytic differentiation of K562 cells by knockdown of the gene. In this study, we examined the effect of HZF1 on the proliferation and apoptosis of K562 cells and identified the possible mechanism for this effect. By lentivirus-mediated gene transfer, we obtained stable K562 transductants with HZF1 overexpression (K562/WPXL-HZF1) and stable control transductants (K562/WPXL). Significantly rapid cell amplification was observed in K562/WPXL-HZF1 cells compared to K562/WPXL cells. The cell cycles of the two transductants were analyzed and the results demonstrated that HZF1 overexpression promoted the S to G2/M phase transition. Additionally, we found that the overexpression of HZF1 slightly inhibits the apoptosis of K562 cells induced by sodium arsenate. Furthermore, using a yeast two-hybrid (Y2H) system we identified the HZF1-interacting proteins and screened 29 potential binding partners of HZF1. Using a co-immunoprecipitation (Co-IP) assay, we confirmed the interaction between HZF1 and the inhibitor of cyclin-dependent kinase (CDK) interacting with cyclin A1 (INCA1), and proved that this interaction leads to the inhibition of INCA1 function, which rescued the activity of CDK2 inhibited by INCA1. In conclusion, our results identified novel functions of the HZF1 gene and revealed a possible mechanism through which HZF1 affects K562 cell proliferation.

[1]  Junwu Zhang,et al.  Identification of the Trans-Activation Domain and the Nuclear Location Signals of Human Zinc Finger Protein HZF1 (ZNF16) , 2010, Molecular biotechnology.

[2]  David J. Segal,et al.  Keep Your Fingers Off My DNA: Protein–Protein Interactions Mediated by C2H2 Zinc Finger Domains , 2008, Cell Biochemistry and Biophysics.

[3]  H. Peng,et al.  Identification and characterization of a novel zinc finger protein (HZF1) gene and its function in erythroid and megakaryocytic differentiation of K562 cells , 2006, Leukemia.

[4]  M. Wiznerowicz,et al.  A versatile tool for conditional gene expression and knockdown , 2006, Nature Methods.

[5]  P. Ji,et al.  Identification of Interaction Partners and Substrates of the Cyclin A1-CDK2 Complex* , 2004, Journal of Biological Chemistry.

[6]  L. Khachigian,et al.  Protein-Protein Interaction between Fli-1 and GATA-1 Mediates Synergistic Expression of Megakaryocyte-Specific Genes through Cooperative DNA Binding , 2004, Molecular and Cellular Biology.

[7]  M. Bhat,et al.  Identification of the Drosophila progenitor of mammalian Krüppel-like factors 6 and 7 and a determinant of fly development. , 2003, Gene.

[8]  Levon M. Khachigian,et al.  Protein-Protein Interaction between Fli-1 and GATA-1 Mediates Synergistic Expression of Megakaryocyte-Specific Genes through Cooperative DNA Binding , 2003, Molecular and Cellular Biology.

[9]  L. Zon,et al.  neptune, a Krüppel-like transcription factor that participates in primitive erythropoiesis in Xenopus , 2001, Current Biology.

[10]  Stephen L. Johnson,et al.  The zebrafish klf gene family. , 2001, Blood.

[11]  J. Bieker.  Krüppel-like Factors: Three Fingers in Many Pies* , 2001, The Journal of Biological Chemistry.

[12]  D. Dang,et al.  The biology of the mammalian Krüppel-like family of transcription factors. , 2000, The international journal of biochemistry & cell biology.

[13]  M. Crossley,et al.  Mammalian Krüppel-like transcription factors: more than just a pretty finger. , 1999, Trends in biochemical sciences.

[14]  M. Lübbert,et al.  Cyclin A1 expression in leukemia and normal hematopoietic cells. , 1999, Blood.

[15]  H P Koeffler,et al.  Characterization of a second human cyclin A that is highly expressed in testis and in several leukemic cell lines. , 1997, Cancer research.

[16]  David O. Morgan,et al.  Principles of CDK regulation , 1995, Nature.

[17]  L. Tsai,et al.  The cdk2 kinase is required for the G1-to-S transition in mammalian cells. , 1993, Oncogene.

[18]  R. Pepperkok,et al.  Cyclin A is required at two points in the human cell cycle. , 1992, The EMBO journal.

[19]  M. Meyerson,et al.  Isolation of the human cdk2 gene that encodes the cyclin A- and adenovirus E1A-associated p33 kinase , 1991, Nature.