[Identification of proteins interacting with androgen receptor- associated coregulator 267-alpha (ARA267-alpha) with the yeast two-hybrid system].
OBJECTIVE
To search for proteins interacting with ARA267-alpha with the yeast two-hybrid system in order to further investigate the function of ARA267-alpha.


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-alpha. 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-alpha-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-alpha-PHD-SET.


CONCLUSIONS
The ARA267-alpha-PHD-SET can interact with several distinct proteins. This suggests that ARA267-alpha is a protein having multiple functions. RanBPM might be a transcriptional factor.
Department of General Surgery, Peking University First Hospital, Beijing 100034, China. wangxin_guo@hotmail.com
Xin
X
Wang
Tie-jun
TJ
Mai
Yi-nong
YN
Niu
Jian-wei
JW
Wang
Ying-lu
YL
Guo
D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019295:Computational Biology; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D020798:Two-Hybrid System Techniques
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C113272:NSD1 protein, human