Hypothermic stress leads to activation of Ras-Erk signaling.

The small GTPase Ras is converted to the active, GTP-bound state during exposure of vertebrate cells to hypothermic stress. This activation occurs more rapidly than can be accounted for by spontaneous nucleotide exchange. Ras-guanyl nucleotide exchange factors and Ras GTPase-activating proteins have significant activity at 0 degrees C in vitro, leading to the hypothesis that normal Ras regulators influence the relative amounts of Ras-GTP and Ras-GDP at low temperatures in vivo. When hypothermic cells are warmed to 37 degrees C, the Raf-Mek-Erk protein kinase cascade is activated. After prolonged hypothermic stress, followed by warming to physiologic temperature, cultured fibroblasts assume a rounded morphology, detach from the substratum, and die. All of these biologic responses are attenuated by pharmacologic inhibition of Mek. Previously, it had been found that low temperature blocks acute growth factor signaling to Erk. In the present study, we found that this block occurs at the level of Raf activation. Temperature regulation of Ras signaling could help animal cells respond appropriately to hypothermic stress, and Ras-Erk signaling can be manipulated to improve the survival of cells in cold storage.

[1]  D. Clawson,et al.  Oncogenes, growth factors and phorbol esters regulate Raf-1 through common mechanisms , 1998, Oncogene.

[2]  F. Hobbs,et al.  Identification of a Novel Inhibitor of Mitogen-activated Protein Kinase Kinase* , 1998, The Journal of Biological Chemistry.

[3]  J. Stone,et al.  RasGRP, a Ras guanyl nucleotide- releasing protein with calcium- and diacylglycerol-binding motifs. , 1998, Science.

[4]  C. Y. Wang,et al.  Requirement of NF-kappaB activation to suppress p53-independent apoptosis induced by oncogenic Ras. , 1997, Science.

[5]  Asim Khwaja,et al.  Matrix adhesion and Ras transformation both activate a phosphoinositide 3‐OH kinase and protein kinase B/Akt cellular survival pathway , 1997, The EMBO journal.

[6]  P. Warne,et al.  Role of Phosphoinositide 3-OH Kinase in Cell Transformation and Control of the Actin Cytoskeleton by Ras , 1997, Cell.

[7]  M. Ewen,et al.  Ras signalling linked to the cell-cycle machinery by the retinoblastoma protein , 1997, Nature.

[8]  M. Schwartz,et al.  Suppression of Integrin Activation: A Novel Function of a Ras/Raf-Initiated MAP Kinase Pathway , 1997, Cell.

[9]  G. Superti-Furga,et al.  Opposite effects of the p52shc/p46shc and p66shc splicing isoforms on the EGF receptor–MAP kinase–fos signalling pathway , 1997, The EMBO journal.

[10]  G. Evan,et al.  Suppression of c-Myc-induced apoptosis by Ras signalling through PI(3)K and PKB , 1997, Nature.

[11]  D. Shalloway,et al.  Cell cycle-dependent activation of Ras , 1996, Current Biology.

[12]  D. Lowy,et al.  Rasp21 sequences opposite the nucleotide binding pocket are required for GRF-mediated nucleotide release. , 1996, Oncogene.

[13]  J. McAnulty,et al.  Effect of hypothermia on intracellular Ca2+ in rabbit renal tubules suspended in UW-gluconate preservation solution. , 1996, Cryobiology.

[14]  Philip R. Cohen,et al.  PD 098059 Is a Specific Inhibitor of the Activation of Mitogen-activated Protein Kinase Kinase in Vitro and in Vivo(*) , 1995, The Journal of Biological Chemistry.

[15]  J. Stone,et al.  RAS signalling is abnormal in a c-raf1 MEK1 double mutant , 1995, Molecular and cellular biology.

[16]  M. Karin,et al.  Cytoskeletal control of gene expression: depolymerization of microtubules activates NF-kappa B , 1995, The Journal of cell biology.

[17]  C. Marshall,et al.  Specificity of receptor tyrosine kinase signaling: Transient versus sustained extracellular signal-regulated kinase activation , 1995, Cell.

[18]  T. Hunter,et al.  Integrin-mediated signal transduction linked to Ras pathway by GRB2 binding to focal adhesion kinase , 1994, Nature.

[19]  C. Gregory,et al.  Regulation of cell survival in burkitt lymphoma: Implications from studies of apoptosis following cold‐shock treatment , 1994, International journal of cancer.

[20]  W. Xie,et al.  The cysteine-rich region of raf-1 kinase contains zinc, translocates to liposomes, and is adjacent to a segment that binds GTP-ras. , 1994, The Journal of biological chemistry.

[21]  R. Davis,et al.  The native structure of the activated Raf protein kinase is a membrane-bound multi-subunit complex. , 1994, The Journal of biological chemistry.

[22]  Mark S. Boguski,et al.  Proteins regulating Ras and its relatives , 1993, Nature.

[23]  J. Stone,et al.  Effector domain mutations dissociate p21ras effector function and GTPase-activating protein interaction , 1993, Molecular and cellular biology.

[24]  P. Warne,et al.  Direct interaction of Ras and the amino-terminal region of Raf-1 in vitro , 1993, Nature.

[25]  Julian Downward,et al.  Epidermal growth factor regulates p21 ras through the formation of a complex of receptor, Grb2 adapter protein, and Sos nucleotide exchange factor , 1993, Cell.

[26]  I. Beletsky,et al.  Apoptosis of murine BW 5147 thymoma cells induced by cold shock , 1992, Journal of cellular physiology.

[27]  M. Hallek,et al.  Granulocyte-macrophage colony-stimulating factor, interleukin-3, and steel factor induce rapid tyrosine phosphorylation of p42 and p44 MAP kinase. , 1992, Blood.

[28]  R. Campos-González,et al.  Temperature-dependent tyrosine phosphorylation of microtubule-associated protein kinase in epidermal growth factor-stimulated human fibroblasts. , 1991, Cell regulation.

[29]  H. Land,et al.  Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. , 1990, Nucleic acids research.

[30]  J H Southard,et al.  Principles of solid-organ preservation by cold storage. , 1988, Transplantation.

[31]  P. W. Hochachka.  Defense strategies against hypoxia and hypothermia. , 1986, Science.

[32]  E. Scolnick,et al.  Viral Harvey ras p21 expressed in Escherichia coli purifies as a binary one-to-one complex with GDP. , 1985, The Journal of biological chemistry.

[33]  W. Topp.  Normal rat cell lines deficient in nuclear thymidine kinase. , 1981, Virology.

[34]  D. Lowy,et al.  Function and regulation of ras. , 1993, Annual review of biochemistry.

[35]  G. Nolan,et al.  Production of high-titer helper-free retroviruses by transient transfection. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[36]  L. Wang,et al.  Effect of low temperature on the cytosolic free Ca2+ in rat ventricular myocytes. , 1991, Cell calcium.

[37]  S. Orrenius,et al.  Calcium‐dependent DNA fragmentation in human synovial cells exposed to cold shock , 1990, FEBS letters.

[38]  R. Tsien,et al.  Requirement for intrinsic protein tyrosine kinase in the immediate and late actions of the EGF receptor , 1987, Nature.