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A glycine-rich RNA-binding protein mediating cold-inducible suppression of mammalian cell growth.

Nishiyama H, Itoh K, Kaneko Y, Kishishita M, Yoshida O, Fujita J - J. Cell Biol. (1997)

Bottom Line: The cirp cDNA encoded an 18-kD protein consisting of an amino-terminal RNAbinding domain and a carboxyl-terminal glycine-rich domain and exhibited structural similarity to a class of stress-induced RNA-binding proteins found in plants.When the culture temperature was lowered from 37 to 32 degrees C, expression of CIRP was induced and growth of BALB/3T3 cells was impaired as compared with that at 37 degrees C.By suppressing the induction of CIRP with antisense oligodeoxynucleotides, this impairment was alleviated, while overexpression of CIRP resulted in impaired growth at 37 degrees C with prolongation of G1 phase of the cell cycle.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Molecular Biology, Faculty of Medicine, Kyoto University, Kyoto 606, Japan.

ABSTRACT
In response to low ambient temperature, mammalian cells as well as microorganisms change various physiological functions, but the molecular mechanisms underlying these adaptations are just beginning to be understood. We report here the isolation of a mouse cold-inducible RNA-binding protein (cirp) cDNA and investigation of its role in cold-stress response of mammalian cells. The cirp cDNA encoded an 18-kD protein consisting of an amino-terminal RNAbinding domain and a carboxyl-terminal glycine-rich domain and exhibited structural similarity to a class of stress-induced RNA-binding proteins found in plants. Immunofluorescence microscopy showed that CIRP was localized in the nucleoplasm of BALB/3T3 mouse fibroblasts. When the culture temperature was lowered from 37 to 32 degrees C, expression of CIRP was induced and growth of BALB/3T3 cells was impaired as compared with that at 37 degrees C. By suppressing the induction of CIRP with antisense oligodeoxynucleotides, this impairment was alleviated, while overexpression of CIRP resulted in impaired growth at 37 degrees C with prolongation of G1 phase of the cell cycle. These results indicate that CIRP plays an essential role in cold-induced growth suppression of mouse fibroblasts. Identification of CIRP may provide a clue to the regulatory mechanisms of cold responses in mammalian cells.

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(a) Comparison of amino acid sequences in the RNA-binding domains of mouse CIRP, human RBM3 (Derry et al., 1995), B.  napus BnGRP10 (Bergeron et al., 1993), A. thaliana Ccr1 and Ccr2 (Carpenter et al., 1994), and human hnRNP G (Soulard et al., 1993).  The consensus sequence for the RNA-binding domain, as determined by Burd and Dreyfuss (1994), is also shown. Dots indicate amino  acids identical to the CIRP sequence. The sequences, RNP1, RNP2, DRET, and MNGKXXDG, are boxed. (b) Structural comparison of  CIRP with its related proteins. The length of each bar reflects the actual length of the sequences. The numbers in the black boxes indicate percent identity of amino acid sequence to CIRP in the CS-RBD (RNP motif). Note that the glycine-rich domain of cyanobacterium Anabaena variabilis RbpA1 (Sato, 1995) is smaller than that of CIRP.
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Figure 2: (a) Comparison of amino acid sequences in the RNA-binding domains of mouse CIRP, human RBM3 (Derry et al., 1995), B. napus BnGRP10 (Bergeron et al., 1993), A. thaliana Ccr1 and Ccr2 (Carpenter et al., 1994), and human hnRNP G (Soulard et al., 1993). The consensus sequence for the RNA-binding domain, as determined by Burd and Dreyfuss (1994), is also shown. Dots indicate amino acids identical to the CIRP sequence. The sequences, RNP1, RNP2, DRET, and MNGKXXDG, are boxed. (b) Structural comparison of CIRP with its related proteins. The length of each bar reflects the actual length of the sequences. The numbers in the black boxes indicate percent identity of amino acid sequence to CIRP in the CS-RBD (RNP motif). Note that the glycine-rich domain of cyanobacterium Anabaena variabilis RbpA1 (Sato, 1995) is smaller than that of CIRP.

Mentions: RNP78 (cirp) contained a 1,264-bp insert with an open reading frame potentially encoding 172 amino acids (Fig. 1). The sequence around the first ATG codon at nucleotide position 81 provided a favorable context for translation initiation (Kozak, 1991), and the 5′ untranslated region contained an in-frame stop codon at position 10. The predicted amino acid sequence displayed two main features: the presence of an amino-terminal CS-RBD and a carboxyl-terminal glycine-rich domain. The CS-RBD of cirp contained consensus sequences of RNP1, RNP2, and a number of other, mostly hydrophobic conserved amino acids interspersed throughout the motif (Fig. 2 a). The carboxyl-terminal part was rich in glycine, serine, arginine, and tyrosine (38.8, 16.4, 19.4, and 10.4%, respectively).


A glycine-rich RNA-binding protein mediating cold-inducible suppression of mammalian cell growth.

Nishiyama H, Itoh K, Kaneko Y, Kishishita M, Yoshida O, Fujita J - J. Cell Biol. (1997)

(a) Comparison of amino acid sequences in the RNA-binding domains of mouse CIRP, human RBM3 (Derry et al., 1995), B.  napus BnGRP10 (Bergeron et al., 1993), A. thaliana Ccr1 and Ccr2 (Carpenter et al., 1994), and human hnRNP G (Soulard et al., 1993).  The consensus sequence for the RNA-binding domain, as determined by Burd and Dreyfuss (1994), is also shown. Dots indicate amino  acids identical to the CIRP sequence. The sequences, RNP1, RNP2, DRET, and MNGKXXDG, are boxed. (b) Structural comparison of  CIRP with its related proteins. The length of each bar reflects the actual length of the sequences. The numbers in the black boxes indicate percent identity of amino acid sequence to CIRP in the CS-RBD (RNP motif). Note that the glycine-rich domain of cyanobacterium Anabaena variabilis RbpA1 (Sato, 1995) is smaller than that of CIRP.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2139845&req=5

Figure 2: (a) Comparison of amino acid sequences in the RNA-binding domains of mouse CIRP, human RBM3 (Derry et al., 1995), B. napus BnGRP10 (Bergeron et al., 1993), A. thaliana Ccr1 and Ccr2 (Carpenter et al., 1994), and human hnRNP G (Soulard et al., 1993). The consensus sequence for the RNA-binding domain, as determined by Burd and Dreyfuss (1994), is also shown. Dots indicate amino acids identical to the CIRP sequence. The sequences, RNP1, RNP2, DRET, and MNGKXXDG, are boxed. (b) Structural comparison of CIRP with its related proteins. The length of each bar reflects the actual length of the sequences. The numbers in the black boxes indicate percent identity of amino acid sequence to CIRP in the CS-RBD (RNP motif). Note that the glycine-rich domain of cyanobacterium Anabaena variabilis RbpA1 (Sato, 1995) is smaller than that of CIRP.
Mentions: RNP78 (cirp) contained a 1,264-bp insert with an open reading frame potentially encoding 172 amino acids (Fig. 1). The sequence around the first ATG codon at nucleotide position 81 provided a favorable context for translation initiation (Kozak, 1991), and the 5′ untranslated region contained an in-frame stop codon at position 10. The predicted amino acid sequence displayed two main features: the presence of an amino-terminal CS-RBD and a carboxyl-terminal glycine-rich domain. The CS-RBD of cirp contained consensus sequences of RNP1, RNP2, and a number of other, mostly hydrophobic conserved amino acids interspersed throughout the motif (Fig. 2 a). The carboxyl-terminal part was rich in glycine, serine, arginine, and tyrosine (38.8, 16.4, 19.4, and 10.4%, respectively).

Bottom Line: The cirp cDNA encoded an 18-kD protein consisting of an amino-terminal RNAbinding domain and a carboxyl-terminal glycine-rich domain and exhibited structural similarity to a class of stress-induced RNA-binding proteins found in plants.When the culture temperature was lowered from 37 to 32 degrees C, expression of CIRP was induced and growth of BALB/3T3 cells was impaired as compared with that at 37 degrees C.By suppressing the induction of CIRP with antisense oligodeoxynucleotides, this impairment was alleviated, while overexpression of CIRP resulted in impaired growth at 37 degrees C with prolongation of G1 phase of the cell cycle.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Molecular Biology, Faculty of Medicine, Kyoto University, Kyoto 606, Japan.

ABSTRACT
In response to low ambient temperature, mammalian cells as well as microorganisms change various physiological functions, but the molecular mechanisms underlying these adaptations are just beginning to be understood. We report here the isolation of a mouse cold-inducible RNA-binding protein (cirp) cDNA and investigation of its role in cold-stress response of mammalian cells. The cirp cDNA encoded an 18-kD protein consisting of an amino-terminal RNAbinding domain and a carboxyl-terminal glycine-rich domain and exhibited structural similarity to a class of stress-induced RNA-binding proteins found in plants. Immunofluorescence microscopy showed that CIRP was localized in the nucleoplasm of BALB/3T3 mouse fibroblasts. When the culture temperature was lowered from 37 to 32 degrees C, expression of CIRP was induced and growth of BALB/3T3 cells was impaired as compared with that at 37 degrees C. By suppressing the induction of CIRP with antisense oligodeoxynucleotides, this impairment was alleviated, while overexpression of CIRP resulted in impaired growth at 37 degrees C with prolongation of G1 phase of the cell cycle. These results indicate that CIRP plays an essential role in cold-induced growth suppression of mouse fibroblasts. Identification of CIRP may provide a clue to the regulatory mechanisms of cold responses in mammalian cells.

Show MeSH
Related in: MedlinePlus