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[Screening and identification of Polo-like kinase 1 as a 14-3-3Zeta-interacting protein]
Jian Du1, Li Cheng, Li-jian Chen
1Department of Biochemistry and Molecular Biology, School of Basic Medicine, Hefei, China. dujane@163.com
Aim:
To screen 14-3-3Zeta-interacting proteins by the yeast two-hybrid system and confirm the interaction between 14-3-3Zeta and Polo-like kinase 1 (Plk1).
Methods:
A bait vector pGBKT7-14-3-3Zeta was constructed to screen the 14-3-3Zeta-interacting proteins from the HeLa cDNA library by yeast two-hybrid system. The interaction between 14-3-3Zeta and Plk1 was further confirmed by immunoprecipitation assay. We also examined the distribution of endogenous 14-3-3Zeta and Plk1 in mitosis using immunofluorescent staining.
Results:
Plk1 was identified as a partner of 14-3-3Zeta protein by the yeast two-hybrid system. Immunoprecipitation assay demonstrated the interaction between 14-3-3Zeta and Plk1. Immunofluorescence indicated 14-3-3Zeta was colocalized with Plk1 at the midbody during cytokinesis.
Conclusion:
Plk1 is a serine/threonine protein kinase that plays multiple critical roles in centrosome maturation, mitotic chromosome segregation, cytokinesis, and the DNA damage response. The interaction between 14-3-3Zeta and Plk1 suggests 14-3-3 family emerges as a novel player in the mitotic regulation.
