Identification of a Protein Interacting with Apoptin from Human Leucocyte cDNA Library by Using Yeast Two-hybrid Screening

SUN Guo-Jing, TONG Xin, DONG Yan, MEI Zhu-Zhong,
SUN Zhi-Xian^*
( Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850,China )

Abstract    To screen the protein interacting with apoptin from human leucocyte cDNA library by using yeast two-hybrid system, four clones interacting with apoptin were identified.One of them was homologous with Nmi (N-Myc interaction protein).Cell co-immunoprecipitation showed that apoptin could bind to Nmi in mammalian cells. Apoptin mutants T1,T2 and T3 lacked the C-terminal 11 AA,33—46 AA and both,respectively.Apoptin mutants T2 and T3 failed to interact with Nmi,suggesting that its 33—46 AA was pivotal for the interaction. Apoptin mutant T1 still interacted with Nmi,suggesting that its C-terminal 11 AA was not essential for the interaction.
Key words    apoptin;yeast two-hybrid;Nmi;co-immunoprecipitation

^*Corresponding author:Tel,86-10-66931218;Fax,86-10-68214653;e-mail,[email protected]