Identification of proteins interacting with androgen receptor- associated coregulator 267-?(ARA267-?) with the yeast two-hybrid system / 北京大学学报(医学版)

ABSTRACT

Objective:To search for proteins interacting with ARA267-? with the yeast two-hybrid system in order to further investigate the function of ARA267-?. Methods:We screened a pretransformed human brain cDNA library with the pGBKT7-PHD-SET recombinant plasmid as a bait which express four PHD(plant homeodomain) and one SET[Su(var)3-9, Enhancer-of-zeste, Trithorax] conserved domains in ARA267-?.The plasmids in positive yeast clones were selectively identified by restriction analysis and DNA sequencing. The interactions were retested by yeast two-hybrid assay. Results: There were about six hundreds positive yeast clones on SD/-Ade/-His/-Leu/-Trp/2.5 mmol/L 3-AT/ X-?-Gal high-stringency selection plates. The pACT2-cDNA plasmids in sixty-five yeast clones were isolated and thirty-five cDNA inserts were sequenced. Sixteen different genes,including DR6(death receptor-6), PIAS3 (protein inhibitor of activated STAT3)and RanBPM(Ran-binding protein in the microtubule-organizing center), were identified after BLAST in GenBank. The yeast two-hybrid retest showed that all but RanBPM were true interactors of ARA267-?-PHD-SET. Conclusion: The ARA267-?-PHD-SET can interact with several distinct proteins. This suggests that ARA267-? is a protein having multiple functions. RanBPM might be a transcriptional factor.