Germ cell-specific proteins interact with the 3' untranslated regions of Prm-1 and Prm-2 mRNA.
The testis-specific mouse protamine genes (Prm-1 and Prm-2) are transcribed in haploid round spermatids, their mRNAs stored as cytoplasmic ribonucleoprotein particles and translated about 1 week later in elongating spermatids. We have compared the in vitro translational efficiencies of deproteinized Prm-1 mRNA isolated from purified populations of germ cells and found that Prm-1 mRNA from round spermatids translates as efficiently as Prm-1 mRNA from elongating spermatids, suggesting that translation of Prm-1 mRNA is normally repressed in round spermatids. Previous studies in transgenic mice have shown that the 3' UTR of Prm-1 mRNA is necessary and sufficient for its translational control (Braun et al., 1989). In this manuscript, we have used an RNA band shift assay to identify an activity, present in cytoplasmic fractions of meiotic spermatocytes and postmeiotic round spermatids, that binds the 3'UTRs of both Prm-1 and Prm-2 mRNA. We have used 3' UTR deletion variants to map the binding site to a 22-nt region within the Prm-1 3' UTR and to a 20-nt region within the Prm-2 3' UTR. uv cross-linking of the RNA band shift activities detected with the Prm-1 and Prm-2 3' UTRs generated the same two RNA/protein complexes of 53 and 55 kDa. The presence of the binding activity in the cell type and subcellular compartment associated with Prm-1 and Prm-2 mRNA storage suggest that the activity may be actively engaged in translational repression of these mRNAs.
Fajardo
M A
MA
Department of Genetics, University of Washington, Seattle 98195.
Butner
K A
KA
Lee
K
K
Braun
R E
RE
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D018507:Gene Expression Regulation, Developmental; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry; D011479:Protamines / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D017384:Sequence Deletion; D013087:Spermatids / Q000502:physiology*; D013737:Testis / Q000378:metabolism
D015345:Oligonucleotide Probes; D011479:Protamines; D012333:RNA, Messenger; D016601:RNA-Binding Proteins