Translocation of hsp-70 and protein synthesis during continuous heating at mild temperatures in HeLa cells.

We have investigated intracellular translocation of hsp-70 and the synthesis of hsp-70 and total protein during heating at moderate temperatures in HeLa cells. When cells were heated at temperatures above 41 degrees C, hsp-70 translocated from the cytoplasm into the nuclei and apparently accumulated in nucleoli within 10 min. At temperatures above 42 degrees C, hsp-70 remained in the nuclei during heating. When cells were heated at 41 degrees C, the hsp-70 which had translocated into the nuclei returned gradually to the cytoplasm during heating. Synthesis of hsp-70 increased to three- to fourfold that in control cells, then decreased to the control level by 6-8 h. Total protein synthesis first decreased to 60% of the control level, then gradually recovered by 4 h. This indicates the acquisition of translational tolerance during heating at 41 degrees C. The return of hsp-70 to the cytoplasm is related to the recovery of total protein synthesis. Similar results were obtained at 42 degrees C heating in heat-induced thermotolerant cells. From these results, it is suggested that translocation of hsp-70 into the nuclei is very important for the recovery of protein synthesis (acquisition of translational tolerance) during heating at moderate temperatures. Also, cycloheximide and puromycin appeared to lower the temperature threshold about 1 degree C with respect to the translocation of hsp-70.

[1]  J. Garrels,et al.  Identification, characterization, and purification of two mammalian stress proteins present in mitochondria, grp 75, a member of the hsp 70 family and hsp 58, a homolog of the bacterial groEL protein. , 1989, The Journal of biological chemistry.

[2]  Y. Lee,et al.  Heat protectors and heat‐induced preferential redistribution of 26 and 70 kDa proteins in chinese hamster ovary cells , 1989, Journal of cellular physiology.

[3]  N. Brot,et al.  A member of the Hsp70 family is localized in mitochondria and resembles Escherichia coli DnaK. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[4]  A. Laszlo,et al.  The relationship of heat-shock proteins, thermotolerance, and protein synthesis. , 1988, Experimental cell research.

[5]  A. Laszlo.  Evidence for two states of thermotolerance in mammalian cells. , 1988, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[6]  G. Blobel,et al.  70K heat shock related proteins stimulate protein translocation into microsomes , 1988, Nature.

[7]  Elizabeth A. Craig,et al.  A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides , 1988, Nature.

[8]  W. Welch,et al.  Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein 70, intermediate filaments, and small nuclear ribonucleoprotein complexes , 1988, The Journal of cell biology.

[9]  W. Welch,et al.  Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heat-shock protein 70 expression , 1988, The Journal of cell biology.

[10]  W. Dewey,et al.  Effect of cycloheximide or puromycin on induction of thermotolerance by sodium arsenite in Chinese hamster ovary cells: Involvement of heat shock proteins , 1987, Journal of cellular physiology.

[11]  W. Welch,et al.  Cellular and biochemical events in mammalian cells during and after recovery from physiological stress , 1986, The Journal of cell biology.

[12]  K. Ohtsuka,et al.  Possible cytoskeletal association of 69,000- and 68,000-dalton heat shock proteins and structural relations among heat shock proteins in murine mastocytoma cells. , 1986, Radiation research.

[13]  H. Pelham.  Speculations on the functions of the major heat shock and glucose-regulated proteins , 1986, Cell.

[14]  S. Munro,et al.  An hsp70-like protein in the ER: Identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein , 1986, Cell.

[15]  K. Ohtsuka,et al.  Intracellular distribution of 73,000 and 72,000 dalton heat shock proteins in HeLa cells. , 1986, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[16]  W. Dewey,et al.  Protection of Chinese hamster ovary cells from hyperthermic killing by cycloheximide or puromycin. , 1986, Radiation research.

[17]  H. Pelham,et al.  Involvement of ATP in the nuclear and nucleolar functions of the 70 kd heat shock protein. , 1985, The EMBO journal.

[18]  W. Welch,et al.  Rapid purification of mammalian 70,000-dalton stress proteins: affinity of the proteins for nucleotides. , 1985, Molecular and cellular biology.

[19]  H. Pelham.  Hsp70 accelerates the recovery of nucleolar morphology after heat shock. , 1984, The EMBO journal.

[20]  J. Subjeck,et al.  Heat shock proteins and protection of proliferation and translation in mammalian cells. , 1984, Cancer research.

[21]  W. Welch,et al.  Nuclear and nucleolar localization of the 72,000-dalton heat shock protein in heat-shocked mammalian cells. , 1984, The Journal of biological chemistry.

[22]  J. M. Velazquez,et al.  hsp70: Nuclear concentration during environmental stress and cytoplasmic storage during recovery , 1984, Cell.

[23]  J. Garrels,et al.  Biochemical characterization of the mammalian stress proteins and identification of two stress proteins as glucose- and Ca2+-ionophore-regulated proteins. , 1983, The Journal of biological chemistry.

[24]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[25]  K. Henle,et al.  Effects of Hyperthermia (45°) on Macromolecular Synthesis in Chinese Hamster Ovary Cells , 1979 .

[26]  E. Gerner,et al.  Induced thermal resistance in HeLa cells , 1975, Nature.

[27]  S. Penman,et al.  Regulation of protein synthesis in HeLa cells: translation at elevated temperatures. , 1969, Journal of molecular biology.

[28]  Michael Ashburner,et al.  Heat shock, from bacteria to man , 1982 .