Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis.
Variable Charge X/Y (VCX/Y) is a human testis-specific gene family that localized on X and Y chromosomes. In this study, VCY protein was expressed in E. coli in the form of glutathione-S-transferase (GST) fusion protein. With the purified fusion protein as antigen, the anti-GST-VCY antibody was generated and the localization of VCY protein in human testis was determined by immunohistochemistry. In the testis seminiferous epithelium, VCY proteins were highly expressed in nuclei of germ cells. Using propidium iodide staining and green fluorescent protein (GFP) tag technologies, VCY and VCX-8r proteins were mainly localized in the nucleoli of COS7 cells. In addition, the colocalization for VCY and VCX-8r in COS7 cells was also observed. With VCY cDNA as bait, a cDNA fragment of acidic ribosomal protein PO was obtained using yeast two-hybrid system. All the information above indicates that VCX/Y protein family might be involved in the regulation of ribosome assembly during spermatogenesis.
Zou
Sheng Wei
SW
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, 5 Dong Dan 3-tiao, Beijing 100005, China.
Zhang
Jian Chao
JC
Zhang
Xiao Dong
XD
Miao
Shi Ying
SY
Zong
Shu Dong
SD
Sheng
Qi
Q
Wang
Lin Fang
LF
D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D018613:Microscopy, Confocal; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D013091:Spermatogenesis / Q000235:genetics*; D013737:Testis / Q000737:chemistry; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D008164:Luminescent Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; C404411:VCX protein, human; C404412:VCY protein, human; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase