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
- Tufts Univ., Medford, MA (USA)
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.
- OSTI ID:
- 5761749
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 88:9; ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PROTAMINES
BIOCHEMICAL REACTION KINETICS
SPERMATOZOA
GENE REGULATION
CROSS-LINKING
CYTOPLASM
MESSENGER-RNA
MICE
POST-TRANSLATION MODIFICATION
SPERMATOGENESIS
ULTRAVIOLET RADIATION
ANIMALS
CELL CONSTITUENTS
CHEMICAL REACTIONS
COAGULANTS
DRUGS
ELECTROMAGNETIC RADIATION
GAMETES
GAMETOGENESIS
GERM CELLS
HEMATOLOGIC AGENTS
HEPARIN ANTAGONISTS
KINETICS
MAMMALS
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
POLYMERIZATION
PROTEINS
RADIATIONS
REACTION KINETICS
RNA
RODENTS
VERTEBRATES
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture