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Dynamic changes in the localization of thermally unfolded nuclear proteins associated with chaperone-dependent protection
- Source :
- Proceedings of the National Academy of Sciences of the United States of America, 98(21), 12038-12043. NATL ACAD SCIENCES
- Publication Year :
- 2001
-
Abstract
- Molecular chaperones are involved in the protection of cells against protein damage through their ability to hold, disaggregate, and refold damaged proteins or their ability to facilitate degradation of damaged proteins. Little is known about how these processes are spatially coordinated in cells. Using a heat-sensitive nuclear model protein luciferase fused to the traceable, heat-stable enhanced green fluorescent protein (N-luc-EGFP), we now show that heat inactivation and insolubilization of luciferase were associated with accumulation of N-luc-EGFP at multiple foci throughout the nucleus. Coexpression of Hsp70, one of the major mammalian chaperones, reduced the formation of these small foci during heat shock. Instead, the heat-unfolded N-luc-EGFP accumulated in large, insoluble foci. Immunofluorescence analysis revealed that these foci colocalized with the nucleoli. Time-lapse analysis demonstrated that protein translocation to the nucleolus, in contrast to the accumulation at small foci, was fully reversible upon return to the normal growth temperature. This reversibility was associated with an increase in the level of active and soluble luciferase. Expression of a carboxyl-terminal deletion mutant of Hsp70(1–543), which lacked chaperone activity, had no effect on the localization of N-luc-EGFP, which suggests that the Hsp70 chaperone activity is required for the translocation events. Our data show that Hsp70 not only is involved in holding and refolding of heat-unfolded nuclear proteins but also drives them to the nucleolus during stress. This might prevent random aggregation of thermolabile proteins within the nucleus, thereby allowing their refolding at the permissive conditions and preventing indirect damage to other nuclear components.
- Subjects :
- Protein Folding
STRESS
Octoxynol
Nucleolus
Recombinant Fusion Proteins
Detergents
Green Fluorescent Proteins
Cell Line
Heating
MAMMALIAN-CELLS
Cricetinae
Heat shock protein
Animals
HEAT-SHOCK PROTEINS
CENTROSOME
HSP70 Heat-Shock Proteins
Luciferase
Nuclear protein
Thermolabile
Luciferases
IN-VIVO
HSP70
Cell Nucleus
Multidisciplinary
biology
Nuclear Proteins
Biological Sciences
RECOVERY
Cell biology
Hsp70
Luminescent Proteins
Solubility
MOLECULAR CHAPERONES
Chaperone (protein)
biology.protein
Protein folding
MACHINERY
ATPASE ACTIVITY
Subjects
Details
- Language :
- English
- ISSN :
- 00278424
- Volume :
- 98
- Issue :
- 21
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Accession number :
- edsair.doi.dedup.....53c267c419433b4cc361cc56c446cfd8