[Determination of the binding site of testis-specific nucleoporin BS-63 to transportin (karopherin beta 2) and the proof of their combination in vitro].
OBJECTIVE
To locate the binding site of testis-specific nucleoporin BS-63 to transportin (karopherin beta 2) and confirm their combination in vitro.


METHODS
Constructed different fragments of C terminal BS-63 was employed to localize the binding site of the testis-specific nucleoporin BS-63 to transportin by yeast two-hybrid system technique pull-down test was used to identify the interaction between the purified expressed fragments of BS-63 0.6 K and transportin in vitro.


RESULTS
BS-63 binding site to transportin was shortened from 1.6 kb to 0.6 kb which included a Ran binding domain (RanBD). SDS-PAGE and Western blot tests confirmed the recombinant purified protein coded by 0.6 kb fragment of BS-63 cDNA could interact with transportin in vitro.


CONCLUSIONS
In germ cells, the function of the testis-specific nucleoporin BS-63 localized at cytoplasmic side of NPC importing cargoes into nuclear may be accomplished by transportin cooperation.
Cai
Y
Y
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Miao
S Y
SY
Wang
L F
LF
D001665:Binding Sites; D006801:Humans; D066298:In Vitro Techniques; D028884:Karyopherins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism*; D013737:Testis / Q000737:chemistry*; D020798:Two-Hybrid System Techniques
D028884:Karyopherins; D028861:Nuclear Pore Complex Proteins; D011994:Recombinant Proteins