Abstract
Nuclear accumulation of heat shock protein (HSP) 72 occurs after cardiac ischemia. This nuclear accumulation of HSP72 with stress occurs in other tissues and species. We postulated that nuclear accumulation of HSP72 was important for the protective effect of HSP72 and that phosphorylation of a single tyrosine (Y524) regulated nuclear accumulation of HSP72. Western blots of immunoprecipitated HSP72 from Cos-1 cells demonstrated that tyrosine becomes phosphorylated after heat shock. Treatment with the tyrosine kinase inhibitor geldanamycin blocked nuclear accumulation of HSP72 with heat shock. Two epitope-tagged constructs were made: M17 converting Y524 to aspartic acid (pseudophosphorylation) and M18 converting Y524 to phenylalanine. When transfected into Cos-1 cells, M17 accumulates more rapidly and M18 less rapidly than wild-type (WT) HSP72 in the nucleus following heat shock. Cells expressing M18 had less viability after heat shock at 43.5°C than other constructs. After heat shock at 45°C, cells expressing M17 had superior survival compared with WT and M18. These data suggest that phosphorylation at Y524 facilitates nuclear accumulation of HSP72 following heat stress, and substitution of aspartic acid at Y524 enhances resistance to heat-shock injury.
| Original language | English (US) |
|---|---|
| Journal | American Journal of Physiology - Heart and Circulatory Physiology |
| Volume | 278 |
| Issue number | 6 47-6 |
| State | Published - 2000 |
| Externally published | Yes |
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Keywords
- Heat shock protein 70
- Heat shock proteins
- Ischemia
- Nuclear localization
ASJC Scopus subject areas
- Physiology
- Physiology (medical)
Cite this
Phosphorylation at tyrosine-524 influences nuclear accumulation of HSP72 with heat stress. / Knowlton, Anne A; Grenier, M.; Kirchhoff, S. R.; Salfity, M.
In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 278, No. 6 47-6, 2000.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Phosphorylation at tyrosine-524 influences nuclear accumulation of HSP72 with heat stress
AU - Knowlton, Anne A
AU - Grenier, M.
AU - Kirchhoff, S. R.
AU - Salfity, M.
PY - 2000
Y1 - 2000
N2 - Nuclear accumulation of heat shock protein (HSP) 72 occurs after cardiac ischemia. This nuclear accumulation of HSP72 with stress occurs in other tissues and species. We postulated that nuclear accumulation of HSP72 was important for the protective effect of HSP72 and that phosphorylation of a single tyrosine (Y524) regulated nuclear accumulation of HSP72. Western blots of immunoprecipitated HSP72 from Cos-1 cells demonstrated that tyrosine becomes phosphorylated after heat shock. Treatment with the tyrosine kinase inhibitor geldanamycin blocked nuclear accumulation of HSP72 with heat shock. Two epitope-tagged constructs were made: M17 converting Y524 to aspartic acid (pseudophosphorylation) and M18 converting Y524 to phenylalanine. When transfected into Cos-1 cells, M17 accumulates more rapidly and M18 less rapidly than wild-type (WT) HSP72 in the nucleus following heat shock. Cells expressing M18 had less viability after heat shock at 43.5°C than other constructs. After heat shock at 45°C, cells expressing M17 had superior survival compared with WT and M18. These data suggest that phosphorylation at Y524 facilitates nuclear accumulation of HSP72 following heat stress, and substitution of aspartic acid at Y524 enhances resistance to heat-shock injury.
AB - Nuclear accumulation of heat shock protein (HSP) 72 occurs after cardiac ischemia. This nuclear accumulation of HSP72 with stress occurs in other tissues and species. We postulated that nuclear accumulation of HSP72 was important for the protective effect of HSP72 and that phosphorylation of a single tyrosine (Y524) regulated nuclear accumulation of HSP72. Western blots of immunoprecipitated HSP72 from Cos-1 cells demonstrated that tyrosine becomes phosphorylated after heat shock. Treatment with the tyrosine kinase inhibitor geldanamycin blocked nuclear accumulation of HSP72 with heat shock. Two epitope-tagged constructs were made: M17 converting Y524 to aspartic acid (pseudophosphorylation) and M18 converting Y524 to phenylalanine. When transfected into Cos-1 cells, M17 accumulates more rapidly and M18 less rapidly than wild-type (WT) HSP72 in the nucleus following heat shock. Cells expressing M18 had less viability after heat shock at 43.5°C than other constructs. After heat shock at 45°C, cells expressing M17 had superior survival compared with WT and M18. These data suggest that phosphorylation at Y524 facilitates nuclear accumulation of HSP72 following heat stress, and substitution of aspartic acid at Y524 enhances resistance to heat-shock injury.
KW - Heat shock protein 70
KW - Heat shock proteins
KW - Ischemia
KW - Nuclear localization
UR - http://www.scopus.com/inward/record.url?scp=0033921550&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033921550&partnerID=8YFLogxK
M3 - Article
C2 - 10843914
AN - SCOPUS:0033921550
VL - 278
JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology
JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology
SN - 1931-857X
IS - 6 47-6
ER -