Heat-shock Treatment-mediated Increase in Transduction by Recombinant Adeno-associated Virus 2 Vectors Is Independent of the Cellular Heat-shock Protein 90

Here we further characterize a number of properties inherent to the thermotolerant cell. In the preceding paper, we showed that the acquisition of the thermotolerant state (by a prior induction of the heat-shock proteins) renders cells translationally tolerant to a subsequent severe heat-shock treatment and thereby results in faster kinetics of both the synthesis and subsequent repression of the stress proteins. Because of the apparent integral role of the 70-kD stress proteins in the acquisition of tolerance, we compared the intracellular distribution of these proteins in both tolerant and nontolerant cells before and after a severe 45 degrees C/30-min shock. In both HeLa and rat embryo fibroblasts, the synthesis and migration of the major stress-induced 72-kD protein into the nucleolus and its subsequent exit was markedly faster in the tolerant cells as compared with the nontolerant cells. Migration of preexisting 72-kD into the nucleolus was shown to be dependent upon heat-shock treatment and independent of active heat-shock protein synthesis. Using both microinjection and immunological techniques, we observed that the constitutive and abundant 73-kD stress protein similarly showed a redistribution from the cytoplasm and nucleus into the nucleolus as a function of heat-shock treatment. We show also that other lesions that occur in cells after heat shock can be prevented or at least minimized if the cells are first made tolerant. Specifically, the heat-induced collapse of the intermediate filament cytoskeleton did not occur in cells rendered thermotolerant. Similarly, the disruption of intranuclear staining patterns of the small nuclear ribonucleoprotein complexes after heat-shock treatment was less apparent in tolerant cells exposed to a subsequent heat-shock treatment.

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Dynamic changes in the structure and intracellular locale of the mammalian low-molecular-weight heat shock protein

Molecular and Cellular Biology ◽

10.1128/mcb.8.12.5059-5071.1988 ◽

1988 ◽

Vol 8 (12) ◽

pp. 5059-5071

Author(s):

A P Arrigo ◽

J P Suhan ◽

W J Welch

Keyword(s):

Molecular Weight ◽

Heat Shock ◽

Heat Shock Protein ◽

Elevated Temperatures ◽

Shock Treatment ◽

Low Molecular Weight ◽

C Cells ◽

Heat Shock Treatment ◽

Soluble Phase ◽

In Cells

Mammalian cells grown at 37 degrees C contain a single low-molecular-weight heat shock (or stress) protein with an apparent mass of 28 kilodaltons (kDa) whose synthesis increases in cells after exposure to elevated temperatures or other forms of physiologic stress. Herein we present data demonstrating that heat shock protein 28 exists in a number of dynamic states depending upon the physiologic state of the cell. Biochemical fractionation of 37 degrees C cells in the absence of nonionic detergent revealed that the 28-kDa protein partitioned approximately equally between the soluble and insoluble fractions. The addition of detergent in the fractionation procedure resulted in all of the protein distributed within the soluble phase. In contrast, in cells first heat shocked and then fractionated in the presence of detergent, most of the 28-kDa protein was found within the insoluble fraction. These biochemical results appeared entirely consistent with indirect immunofluorescence experiments, demonstrating that the 28-kDa protein resided within the perinuclear region of 37 degrees C cells in close proximity to the Golgi complex. After heat shock treatment, the 28-kDa protein relocalized within the nucleus and resisted detergent extraction. The extent of 28-kDa protein redistribution into the nucleus and its detergent insolubility increased as a function of the severity of the heat shock treatment. With time of recovery from the heat treatment there occurred a gradual return of the 28-kDa protein into the detergent-soluble phase. Concomitant with these changes in 28-kDa protein solubility was a corresponding change in the apparent size of the protein as determined by gel filtration. While at 37 degrees C cells the protein exhibited a mass of 200 to 800 kDa; after heat shock the protein assumed sizes of 2 MDa or greater. Using immunoelectron microscopy, we show an accumulation of these aggregates of 28-kDa protein within the nucleus. Finally, we show that the heat-dependent redistribution of the 28-kDa protein from the cytoplasm into the nucleus was greatly diminished when the cells were first rendered thermotolerant, and we suggest that this simple assay (i.e., 28-kDa protein detergent solubility) may prove useful in evaluating the thermotolerant status of a cell or tissue.

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32. Adeno-Associated Virus 2-Mediated Gene Transfer: Role of Cellular Heat-Shock Protein 90 in Transgene Expression

Molecular Therapy ◽

10.1016/s1525-0016(16)40474-0 ◽

2003 ◽

Vol 7 (5) ◽

pp. S13

Keyword(s):

Heat Shock ◽

Gene Transfer ◽

Heat Shock Protein ◽

Transgene Expression ◽

Heat Shock Protein 90 ◽

Adeno Associated Virus

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Dynamic changes in the structure and intracellular locale of the mammalian low-molecular-weight heat shock protein.

Molecular and Cellular Biology ◽

10.1128/mcb.8.12.5059 ◽

1988 ◽

Vol 8 (12) ◽

pp. 5059-5071 ◽

Cited By ~ 244

Author(s):

A P Arrigo ◽

J P Suhan ◽

W J Welch

Keyword(s):

Molecular Weight ◽

Heat Shock ◽

Heat Shock Protein ◽

Elevated Temperatures ◽

Shock Treatment ◽

Low Molecular Weight ◽

C Cells ◽

Heat Shock Treatment ◽

Soluble Phase ◽

In Cells

Mammalian cells grown at 37 degrees C contain a single low-molecular-weight heat shock (or stress) protein with an apparent mass of 28 kilodaltons (kDa) whose synthesis increases in cells after exposure to elevated temperatures or other forms of physiologic stress. Herein we present data demonstrating that heat shock protein 28 exists in a number of dynamic states depending upon the physiologic state of the cell. Biochemical fractionation of 37 degrees C cells in the absence of nonionic detergent revealed that the 28-kDa protein partitioned approximately equally between the soluble and insoluble fractions. The addition of detergent in the fractionation procedure resulted in all of the protein distributed within the soluble phase. In contrast, in cells first heat shocked and then fractionated in the presence of detergent, most of the 28-kDa protein was found within the insoluble fraction. These biochemical results appeared entirely consistent with indirect immunofluorescence experiments, demonstrating that the 28-kDa protein resided within the perinuclear region of 37 degrees C cells in close proximity to the Golgi complex. After heat shock treatment, the 28-kDa protein relocalized within the nucleus and resisted detergent extraction. The extent of 28-kDa protein redistribution into the nucleus and its detergent insolubility increased as a function of the severity of the heat shock treatment. With time of recovery from the heat treatment there occurred a gradual return of the 28-kDa protein into the detergent-soluble phase. Concomitant with these changes in 28-kDa protein solubility was a corresponding change in the apparent size of the protein as determined by gel filtration. While at 37 degrees C cells the protein exhibited a mass of 200 to 800 kDa; after heat shock the protein assumed sizes of 2 MDa or greater. Using immunoelectron microscopy, we show an accumulation of these aggregates of 28-kDa protein within the nucleus. Finally, we show that the heat-dependent redistribution of the 28-kDa protein from the cytoplasm into the nucleus was greatly diminished when the cells were first rendered thermotolerant, and we suggest that this simple assay (i.e., 28-kDa protein detergent solubility) may prove useful in evaluating the thermotolerant status of a cell or tissue.

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Stress Dose-dependent Suppression of Heat Shock Protein Gene Expression by Inhibiting Protein Synthesis during Heat Shock Treatment.

Cell Structure and Function ◽

10.1247/csf.22.7 ◽

1997 ◽

Vol 22 (1) ◽

pp. 7-13 ◽

Cited By ~ 13

Author(s):

Satoshi Mizuno ◽

Ayako Ishii ◽

Yuko Murakami ◽

Hisayoshi Akagawa

Keyword(s):

Gene Expression ◽

Protein Synthesis ◽

Heat Shock ◽

Heat Shock Protein ◽

Protein Gene ◽

Shock Treatment ◽

Heat Shock Treatment ◽

Heat Shock Protein Gene ◽

Dose Dependent ◽

Stress Dose

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Expression of heat shock protein genes Aaehsp26, Aaehsp83 and Aaehsc70 in response to thermal stress in Aedes aegypti larvae

Dhaka University Journal of Biological Sciences ◽

10.3329/dujbs.v30i2.54649 ◽

2021 ◽

Vol 30 (2) ◽

pp. 233-241

Author(s):

Hafisha Khatun Anee ◽

Ashfaqul Muid Khandaker ◽

Rowshan Ara Begum ◽

Reza Md Shahjahan

Keyword(s):

Climate Change ◽

Heat Shock ◽

Heat Shock Protein ◽

Abiotic Factors ◽

Shock Treatment ◽

Heat Shock Treatment ◽

Quantitative Real Time Pcr ◽

Physiological Processes ◽

Factors Influencing ◽

Heat Shock Protein Genes

Climate change is responsible to a certain extent for the occurrence and spread of arboviral pathogens worldwide. Temperature is one of the crucial abiotic factors influencing the physiological processes of mosquitoes. Several genes of heat shock protein (AaeHsp26, AaeHsp83, and AaeHsc70) families are known to be expressed in mosquitoes, which aid in overcoming stress induced by elevated temperature. In this study, the relative expression of heat shock protein genes has been examined using Quantitative Real-time PCR (qPCR). The temperatures used for heat shock treatment were 27(control), 37, and 42°C for 1 hour heat shock period and applied to 3rd instar larvae. Significant up-regulation has been seen at 37, and 42°C. The highest expression level, about 82.43 fold, was reported for the AaeHsc70 gene at 42°C followed by 78.36 fold for AaeHsp26 at 37°C and 4.79 fold for AaeHsp83 at 42°C. The current study has shown that HSPs are important markers of stress and may function as critical proteins to protect and enhance the survival of Ae. aegypti larvae and pupae. Biological implications of these findings could impact the vector competencies Dhaka Univ. J. Biol. Sci. 30(2): 233-241, 2021 (July)

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