Elevated levels of the polyadenylation factor CstF 64 enhance formation of the 1kB Testis brain RNA-binding protein (TB-RBP) mRNA in male germ cells.
The single copy mouse Testis Brain RNA-Binding Protein (TB-RBP) gene encodes three mRNAs of 3.0, 1.7, and 1.0 kb which only differ in their 3' UTRs. The 1 kb TB-RBP mRNA predominates in testis, while somatic cells preferentially express the 3.0 kb TB-RBP mRNA. Here we show that the 1 kb mRNA is translated several-fold more efficiently than the 3 kb TB-RBP in rabbit reticulocyte lysates and cells with elevated levels of the 1 kB TB-RBP mRNA express high levels of TB-RBP. To determine if the cleavage stimulatory factor CstF 64 can modulate the alternative splicing of the TB-RBP pre-mRNA and therefore TB-RBP expression, CstF 64 levels and binding to alternative polyadenylation sites were examined. CstF 64 is abundant in the testis and preferentially binds to a distal site in the TB-RBP pre-mRNA that produces the 3 kb TB-RBP. Moreover, upregulation or overexpression of CstF 64 increases the poly(A) site selection for the 1 kb TB-RBP mRNA. We propose that the level of the polyadenylation factor CstF 64 modulates the level of TB-RBP synthesis in male germ cells by an alternative processing of the TB-RBP pre-mRNA.
Center for Research on Reproduction and Women's Health and Department of Obstetrics and Gynecology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
V M
VM
Chennathukuzhi
S
S
Lefrancois
C R
CR
Morales
V
V
Syed
N B
NB
Hecht
D020413:3' Untranslated Regions / Q000378:metabolism*; D016475:3T3 Cells; D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry; D002467:Cell Nucleus / Q000378:metabolism; D002474:Cell-Free System; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D007124:Immunoenzyme Techniques; D008099:Liver / Q000378:metabolism; D008297:Male; D008540:Meiosis; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009206:Myocardium / Q000378:metabolism; D009928:Organ Specificity; D011061:Poly A / Q000378:metabolism*; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000502:physiology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000502:physiology; D012156:Reticulocytes / Q000378:metabolism; D012708:Sertoli Cells / Q000737:chemistry; D013087:Spermatids / Q000378:metabolism* / Q000648:ultrastructure; D013090:Spermatocytes / Q000378:metabolism* / Q000648:ultrastructure; D013154:Spleen / Q000378:metabolism; D013737:Testis / Q000737:chemistry; D014162:Transfection; D039221:mRNA Cleavage and Polyadenylation Factors
D020413:3' Untranslated Regions; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D008869:Microtubule-Associated Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C490551:STRBP protein, human; C095563:TSN protein, human; C498303:Tsn protein, mouse; D039221:mRNA Cleavage and Polyadenylation Factors; D011061:Poly A