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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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Effects of antisense (As) ODN on the cold-induction of  p18cirp. BALB/3T3 cells were incubated at the indicated temperatures for 12 h in the presence of vehicle alone, As, or sense (Sn)  ODNs (0.5 μM). Note partial suppression of the cold-induced  p18cirp expression in the presence of antisense ODN.
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Figure 9: Effects of antisense (As) ODN on the cold-induction of p18cirp. BALB/3T3 cells were incubated at the indicated temperatures for 12 h in the presence of vehicle alone, As, or sense (Sn) ODNs (0.5 μM). Note partial suppression of the cold-induced p18cirp expression in the presence of antisense ODN.

Mentions: After a temperature shift from 37 to 32°C, the growth of BALB/3T3 fibroblasts was impaired (Table I). To assess whether the induction of the cirp expression is related to the observed impairment of growth, we examined the effects of ODN antisense to cirp mRNA. The presence of antisense ODN partially inhibited the induction of p18cirp by temperature downshift (Fig. 9). Concomitantly, the growth impairment at 32°C was partially alleviated (Fig. 10), suggesting that the induction of p18cirp is necessary for growth suppression by cold stress.


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)

Effects of antisense (As) ODN on the cold-induction of  p18cirp. BALB/3T3 cells were incubated at the indicated temperatures for 12 h in the presence of vehicle alone, As, or sense (Sn)  ODNs (0.5 μM). Note partial suppression of the cold-induced  p18cirp expression in the presence of antisense ODN.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2139845&req=5

Figure 9: Effects of antisense (As) ODN on the cold-induction of p18cirp. BALB/3T3 cells were incubated at the indicated temperatures for 12 h in the presence of vehicle alone, As, or sense (Sn) ODNs (0.5 μM). Note partial suppression of the cold-induced p18cirp expression in the presence of antisense ODN.
Mentions: After a temperature shift from 37 to 32°C, the growth of BALB/3T3 fibroblasts was impaired (Table I). To assess whether the induction of the cirp expression is related to the observed impairment of growth, we examined the effects of ODN antisense to cirp mRNA. The presence of antisense ODN partially inhibited the induction of p18cirp by temperature downshift (Fig. 9). Concomitantly, the growth impairment at 32°C was partially alleviated (Fig. 10), suggesting that the induction of p18cirp is necessary for growth suppression by cold stress.

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