Cytoplasmic Protein Binding to Highly Conserved Sequences in the 3' Untranslated Region of Mouse Protamine 2 mRNA, a Translationally Regulated Transcript of Male Germ Cells

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 88; no. 9; pp. 3584 - 3588
• Main Authors: KWON, Y. K, HECHT, N. B
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.05.1991; National Acad Sciences
• Subjects: 560120 - Radiation Effects on Biochemicals, Cells, & Tissue Culture; ANIMALS; Antisense oligonucleotides; Base Sequence; Binding Sites; BIOCHEMICAL REACTION KINETICS; Biological and medical sciences; CELL CONSTITUENTS; CHEMICAL REACTIONS; COAGULANTS; Conserved sequences; CROSS-LINKING; CYTOPLASM; Cytoplasm - metabolism; DRUGS; ELECTROMAGNETIC RADIATION; Ferritins; Fundamental and applied biological sciences. Psychology; GAMETES; GAMETOGENESIS; Gels; Gene Expression Regulation; GENE REGULATION; GERM CELLS; HEMATOLOGIC AGENTS; HEPARIN ANTAGONISTS; KINETICS; Male; MAMMALS; MESSENGER-RNA; MICE; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; NUCLEIC ACIDS; NUCLEOPROTEINS; Oligonucleotides; ORGANIC COMPOUNDS; POLYMERIZATION; POST-TRANSLATION MODIFICATION; PROTAMINES; Protamines - genetics; Protein Binding; Protein Biosynthesis; PROTEINS; RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT; RADIATIONS; REACTION KINETICS; Regulatory Sequences, Nucleic Acid; Ribonucleoproteins - physiology; RNA; RNA, Messenger - genetics; RNA, Messenger - metabolism; RODENTS; SPERMATOGENESIS; SPERMATOZOA; Testis - physiology; Translation. Translation factors. Protein processing; ULTRAVIOLET RADIATION; Ultraviolet Rays; Untranslated regions; VERTEBRATES; Ligand binding; Translation; Spermiogenesis; 3' terminal-Sequence; Rodentia; Gel retardation technique; Testicle; Vertebrata; Regulation(control); Mammalia; Messenger RNA; Mouse; Protamine; Molecular complex; Nuclear protein
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.88.9.3584

The expression of the protamines, the predominant nuclear proteins of mammalian spermatozoa, is regulated translationally during male germ-cell development. The 3 ' untranslated region (UTR) of protamine 1 mRNA has been reported to control its time of translation. To understand the mechanisms controlling translation of the protamine mRNAs, we have sought to identify cis elements of the 3 ' UTR of protamine 2 mRNA that are recognized by cytoplasmic factors. From gel retardation assays, two sequence elements are shown to form specific RNA-protein complexes. Protein binding sites of the two complexes were determined by RNase T1 mapping, by blocking the putative binding sites with antisense oligonucleotides, and by competition assays. The sequences of these elements, located between nucleotides +537 and +572 in protamine 2 mRNA, are highly conserved among postmeiotic translationally regulated nuclear proteins of the mammalian testis. Two closely linked protein binding sites were detected. UV-crosslinking studies revealed that a protein of about 18 kDa binds to one of the conserved sequences. These data demonstrate specific protein binding to a highly conserved 3' UTR of translationally regulated testicular mRNA.