CIRP2, a major cytoplasmic RNA-binding protein in Xenopus oocytes.
In an attempt to isolate mRNA-binding proteins we fractionated Xenopus oocyte lysate by oligo(dT)-cellulose chromatography. A 20 kDa protein was the major component of the eluate. cDNA cloning revealed that this protein is a Xenopus homolog of the cold-inducible RNA-binding protein (CIRP) which was originally identified in mammalian cells as a protein that is overexpressed upon a temperature downshift. This Xenopus protein, termed here xCIRP2, is highly expressed in ovary, testis and brain in adult Xenopus tissues. In oocytes it is predominantly localized in the cytoplasm. By biochemical fractionation we provide evidence that xCIRP2 is associated with ribosomes, suggesting that it participates in translational regulation in oocytes. Microinjection of labeled mRNA into oocytes followed by UV cross-linking of the oocyte lysate led to identification of two major RNA-binding activities. Immunoprecipitation of the RNA-binding proteins demonstrated that one is xCIRP2 and that the other contains FRGY2. FRGY2, which is one of the principal constituents of mRNA storage particles involved in translational masking of maternal mRNA, has an RNA-binding domain conserved to those of bacterial cold shock proteins. Possible implications of the highly abundant expression in oocytes of cold shock RNA-binding proteins of both eukaryotic and prokaryotic types are discussed.
Matsumoto
K
K
RIKEN (The Institute of Physical and Chemical Research), Wako, Saitama 351-0198, Japan. matsumok@postman.riken.go.jp
Aoki
K
K
Dohmae
N
N
Takio
K
K
Tsujimoto
M
M
D000595:Amino Acid Sequence; D000818:Animals; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D008297:Male; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D011485:Protein Binding; D012313:RNA / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014981:Xenopus / Q000235:genetics*; D029867:Xenopus Proteins
C491389:CIRP2 protein, Xenopus; D018076:DNA, Complementary; D016601:RNA-Binding Proteins; D029867:Xenopus Proteins; D012313:RNA