Identification of target messenger RNA substrates for mouse RBMY.
Rbmy gene encodes a RNA-binding protein and its expression is limited to the nuclei of germ cells. Previous studies indicate that RBMY may function in pre-mRNA processing during spermatogenesis, although its precise target mRNAs remain unclear. By using specific nucleic acids associated with proteins and immunoprecipitation techniques, we have identified 12 potential target mRNAs bound by mouse RBMY protein from testis. We detect that both mRbmy-1 and mRbmy-2 transcripts co-exist in mouse testis and they differ mainly in the 5'UTR. Importantly, our result shows that mRBMY protein can bind to one of its own transcripts, mRbmy-2, suggesting that mRBMY may affect alternative splicing or regulate the expression of its own gene. Using electrophoretic mobility shift assay, we demonstrated that mRBMY protein can bind to the testis and sperm-specific spa17 mRNA and that the binding domain contains rich oligo(A), suggesting that mRBMY protein may have high affinity to oligo(A) rich sequences. In conclusion, the identification of RBMY target mRNAs will be helpful to further explore the biological function of RBMY in spermatogenesis.
Zeng
Mei
M
Department of Medical Genetics, West China Hospital, Sichuan University, Renmin Nanlu, Section 3 #17, Chengdu 610041, PR China.
Sun
Huaqin
H
Chen
Shu
S
Wang
Xinying
X
Yang
Yuan
Y
Liu
Yunqiang
Y
Tao
Dachang
D
Yang
Zhirong
Z
Zhang
Sizhong
S
Ma
Yongxin
Y
D017398:Alternative Splicing / Q000502:physiology; D000818:Animals; D001665:Binding Sites; D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D019411:Clinical Laboratory Techniques; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011061:Poly A / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012322:RNA Precursors / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013737:Testis / Q000196:embryology / Q000378:metabolism
D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C501500:Rbmy protein, mouse; C490357:Spa17 protein, mouse; D011061:Poly A