Specific binding of a 47-kilodalton protein to the 3' untranslated region of rat transition protein 2 messenger ribonucleic acid.
The synthesis of transition protein (TNP) 2, one of the predominant nuclear proteins of mammalian spermatids, was shown to be posttranscriptionally regulated, by storing the untranslated mRNA for about 3-5 days in the cytoplasm of differentiating spermatids. It has been proposed that binding of a cytoplasmic protein to a conserved motif of 8 nucleotides (nt) in the 3' untranslated region (3'UTR) of TNP2 mRNA is involved in this translational control mechanism. In this report, we show that deletion or variation of the conserved 8-nt motif (GCCAT-CAC) in rat TNP2-3'UTR abolishes the capacity of the in vitro-transcribed RNA to reconstitute specific RNA-protein complexes in RNA bandshift assays. Using Northwestern analysis, we identified specific binding to the TNP2-3'UTR of four proteins of 45, 47, 49, and 60 kDa, all of which are stage-specifically regulated in male germ cell differentiation. Deletion of the 8-nt motif in the 3'UTR specifically prevented binding of the 47-kDa protein, the interaction of which is thought to be mediated by the RNA secondary structure. Analysis of the RNA secondary structure revealed that the 8-nt motif is an essential element of a specific stem-loop structure that is predicted for rat wild-type TNP2-3'UTR. Therefore we assume that the 47-kDa protein plays an important role in specific RNA-protein complex formation of rat TNP2-3'UTR that may be a central event in the translational control of rat TNP2 mRNA.
Schlicker
M
M
Institute of Human Genetics, University of Göttingen, Germany. mschlic@gwdg.de
Reim
K
K
Schlüter
G
G
Engel
W
W
D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002868:Chromosomal Proteins, Non-Histone / Q000096:biosynthesis; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins; D005854:Germ Cells / Q000502:physiology; D008297:Male; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000096:biosynthesis*; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D014158:Transcription, Genetic / Q000502:physiology
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C489504:Tnp2 protein, rat; C053886:spermatid transition proteins; D000645:Ammonium Sulfate