Identification of FANCA as a protein interacting with centromere-associated protein E.

This study sought to isolate and identify proteins that interact with centromere-associated protein E (CENPE), provide new clues for exploring the function of CENP-E in cell cycle control and the pathogenesis of tumor. Yeast two-hybrid screen and regular molecular biologic techniques were undertaken to screen human HeLa cDNA library with ...
the kinetochore binding domain of CENP-E. The bait from the C-terminus of CENP-E was created by subcloning methods to find out optimal candidate proteins that interact with the kinetochore binding domain of CENP-E. Eight novel CENP-E interacting proteins including Homo sapiens Fanconi anemia complementation group A (FANCA) were obtained. In yeast two-hybrid assay, the N-terminal 260 amino acids of FANCA were found to be necessary and sufficient for the interaction with the C-terminus of CENP-E. The interaction was confirmed by in vitro glutathione S-transferase pull-down assay and in vivo coimmunoprecipitation assay. Our finding of the interaction of CENP-E with FANCA demonstrates that CENP-E and FANCA may play important roles in the functional regulation of the mitotic checkpoint signal pathway.
Mesh Terms:
Chromosomal Proteins, Non-Histone, Fanconi Anemia, Genetic Predisposition to Disease, Glutathione Transferase, Hela Cells, Humans, Protein Interaction Domains and Motifs, Tumor Cells, Cultured, Two-Hybrid System Techniques
Acta Biochim. Biophys. Sin. (Shanghai)
Date: Oct. 01, 2009
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