Hsp27 protects the cytoskeleton and nucleus from the effects of 42 degrees C at pH 6.7 in CHO cells adapted to growth at pH 6.7.
CHO cells are normally sensitized to hyperthermia by acidic pH. However, CHO cells adapted to growth in pH 6.7 medium become less sensitive to heat killing at the reduced pH. The adapted cells maintain their ability to develop thermotolerance at pH 6.7 and their steady state intracellular pH is elevated. Furthermore, the small molecular weight stress chaperone, hsp27, is elevated in unheated cells maintained at pH 6.7. This report documents that the cytoskeletal and nuclear components of the low pH adapted CHO cells are resistant to 42 degrees C-induced collapse and protein accretion, respectively. Hyperthermia induced a perinuclear collapse of the microtubular cytoskeleton and an increase in the amount of insoluble protein associated with the nuclei and nuclear matrix fractions in the control cells heated at pH 7.3 or heated after acute acidification to pH 6.7. Protection from these effects was observed in the low pH adapted cells heated at pH 6.7. Hsp70 does not appear to play a dominant role in the response of the adapted cells to 42 degrees C. The induction of hsp70 during heating is abrogated by pH 6.7 in cells cultured at either pH 7.3 or pH 6.7. The resistance of the microtubular cytoskeleton to perinuclear collapse and the absence of protein aggregation in the nucleus during 42 degrees C may be due to the elevated levels of hsp27 both before heating and during the heat treatment. In summary, the phenotype of CHO cells adapted to growth at low pH includes resistance of the cytoskeleton to 42 degrees C-induced perinuclear collapse and resistance to 42 degrees C-induced aggregation of nuclear proteins, in addition to the reduction in heat cytotoxicity, upregulation of intracellular pH and upregulation of hsp27.
Coss
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RA
Department of Radiation Oncology, Kimmel Cancer Center, Thomas Jefferson University, 111 S. 11th Street, Philadelphia, PA 19107-5097, USA. ronald.coss@mail.tju.edu
Sedar
A W
AW
Sistrun
S S
SS
Storck
C W
CW
Wang
P H
PH
Wachsberger
P R
PR
D000222:Adaptation, Physiological; D000818:Animals; D016466:CHO Cells; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D006224:Cricetinae; D003599:Cytoskeleton / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis; D006358:Hot Temperature / Q000009:adverse effects*; D006863:Hydrogen-Ion Concentration; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D015530:Nuclear Matrix / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism
D018840:HSP70 Heat-Shock Proteins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins