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°C, expression of CIRP was induced and growth of BALB/3T3 cells was impaired as compared with that at 37°C. By suppressing the induction of CIRP with antisense oligodeoxynucleotides, this impairment was alleviated, while overexpression of CIRP resulted in impaired growth at 37°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.

This finding provides new evidence that bFGF plays an important role in mediating collateral growth in humans.

Our previous molecular epidemiologic study with gene probes (H. Shirai, H. Ito, T. Hirayama, Y. Nakamoto, N. Nakabayashi, K. Kumagai, Y. Takeda, and M. Nishibuchi, Infect. Immun. 58:3568-3573, 1990) demonstrated that the gene (trh) encoding a thermostable direct hemolysin-related hemolysin was strongly associated with clinical strains of Vibrio parahaemolyticus. Strain-to-strain variation in the intensities of the hybridization signals observed in the above study also suggested that the trh genes in different strains may have significantly divergent nucleotide sequences. To assess the public health significance of the rare environmental strains which exhibited very weak hybridization signals with the trh gene-specific DNA probe, the trh-like sequence was cloned from one of the environmental strains and the nucleotide sequence was determined in this study. A hemolysin gene (trh2) which was 84% homologous to the trh gene (newly named trh1) and 54.8 to 68.8% homologous to the genes (tdh) encoding thermostable direct hemolysins was detected in the cloned sequence. The trh2 gene product showed a profile of hemolytic activities against various animal erythrocytes different from that of the trh1 gene product. The trh2 gene product was antigenically related (partially identical) to the trh1 and tdh gene products. DNA colony blot and Southern blot hybridization analyses with trh1- and trh2-specific DNA probes showed that the trh1 probe-positive strains exhibiting hybridization signals with varying intensities could be clustered into trh1 and trh2 subgroups. In addition, hybridization analysis with oligonucleotide probes demonstrated significant strain-to-strain variation in the trh1 and trh2 gene sequences.(ABSTRACT TRUNCATED AT 250 WORDS)

We isolated a novel hsp110-related gene, apg-1, from a testis cDNA library. The apg-1 transcripts were constitutively expressed in the testicular germ cells and, in some degree, most tissues examined. In a mouse TAMA26 Sertoli cell line, apg-1 transcripts were induced in 2 h by a temperature shift from 32 to 39°C, but not by a shift from 37 to 42°C, the traditional heat stress, or a shift from 32 to 42°C. The heat response pattern of hsp110 expression was similar to that of apg-1. Although induction of a hsp70 transcript was observed in 2 h by a shift from 32 to 39°C, the induction was more apparent by a shift from 37 to 42°C or from 32 to 42°C. Essentially similar differential response patterns were observed among these genes in NIH/3T3 fibroblasts as well. The nuclear run-on assay and the native gel mobility shift assay demonstrated that, by the 32 to 39°C temperature shift, the apg-1 gene was transcriptionally activated, and heat shock factor 1 bound to the heat shock elements in the 5 -flanking region of the apg-1 gene. These results demonstrated that expressions of apg-1, hsp110, and hsp70 could be heat-induced at a temperature lower than the traditional elevated temperatures in somatic cells of both testis and nontestis origin and suggest that the mechanisms regulating the transcript levels of apg-1 and hsp110 are different from those of hsp70. Furthermore, the constitutive expression in germ cells suggests that APG-1 plays a specific role in spermatogenesis as well as in stress response.Prokaryotic and eukaryotic organisms respond to elevated temperatures by synthesizing a distinct set of proteins termed heat shock proteins (HSPs) 1 (1). Anoxia, ethanol, radiation, inflammation, and certain heavy metal ions also induce HSPs in the cells. An early and long-standing assumption regarding the heat shock response was that the HSPs protected cells from the toxic effects of heat and other stresses. Subsequently, a series of studies revealed that HSPs are also present in cells at normal temperatures. Now members of the HSP family are established as molecular chaperones, assisting in the folding and unfolding, assembly and disassembly, and transport of various proteins (2-5). HSPs are also shown to interact with mutant p53 and p60 v-src (6 -8), suggesting their involvement in cell cycle regulation.Spermatogenesis begins at puberty and consists of three steps: the mitotic proliferation of the spermatogonia, meiosis at the spermatocyte stage, and the distinct cellular structural changes of the spermatids. Unlike somatic cells, the male germ cells are easily damaged at the body cavity temperature (9), indicating the presence of a differential heat sensitivity between somatic cells and germ cells. Sarge (10) recently reported that the temperature threshold for induction of HSP72 encoded by the hsp70 gene was lower in male germ cells than in somatic cells. To date, several HSPs have been found to be constitutively expressed in germ cells at specific stages of development. Two HSP70-related genes, hsp70.2 and hsc70t, ar...

The byline should read as shown above. In addition, permission to cite the unpublished data by Adduci et al. (reference 1) was not obtained.