Molecular cloning of sequences encoding the human heat-shock proteins and their expression during hyperthermia (original) (raw)
Related papers
Hyperthermia and the heat-shock proteins of HeLa cells
British journal of cancer, 1982
When HeLa cells are subject to hyperthermia, the synthesis of specific heat-shock proteins (HSP) is induced under a variety of thermal conditions. HSP synthesis does not occur at temperatures above 43 degrees C but requires return to a culture temperature of 37 degrees C. Maximal induction appears to be achieved if a brief hyperthermia treatment (10 min, 45 degrees -46 degrees C) is followed by 2 h "development" at 37 degrees C. The induction process requires transcription but not DNA replication, and general cell metabolism is probably also required, as induction does not occur if the heat-treated cells are returned to 4 degrees (rather than 37 degrees C) for development. A small proportion of the HSPs of 72-74 Kd are found in nuclei, but do not appear to bind to DNA. The bulk of these proteins, as well as those at 100 Kd, are cytoplasmic, but none are preferentially associated with mitochondria. Increased synthesis of the 100Kd and 72-74 Kd HSPs was also triggered by pre...
Constitutively expressed rat mRNA encoding a 70-kilodalton heat-shock-like protein
Molecular and cellular biology, 1985
A nearly full-length cDNA clone isolated from the rat pheochromocytoma cell line, PC12, revealed extensive nucleotide sequence similarity between the rat cDNA and the Drosophila melanogaster hsp70 gene. The rat recombinant clone encodes a 71,000-dalton protein that is 70% identical with the dipteran hsp70 protein. Remarkably, a truncated segment of this cDNA clone was originally isolated by immunoreactivity with antisera raised to catecholamine-synthesizing enzymes, suggesting that this heat shock protein and these catecholamine enzymes shared antigenic determinants. The rat hsp70-related mRNA is responsible for the production of a constitutive hsp70 protein, because it is present in abundant amounts in various tissues at normal growth temperatures and is only minimally induced by hyperthermia. The rat hsp70-related sequence is part of a multigene family that extends across species to mice and humans.
Isolation and functional analysis of a human 70,000-dalton heat shock protein gene segment
Proceedings of the National Academy of Sciences, 1985
A human 70-kDa heat shock protein (hsp7O) gene segment has been isolated. The segment contains 3.15 kilobase pairs (kbp) of 5' nontranscribed sequence, an RNA leader of 119 bp, and a protein-coding region of 741 bp. The human protein sequence shows a high degree of homology to hsp7O sequences from other species. Expression experiments in Xenopus oocytes and mammalian cells indicate that a region that includes only 105 bp of 5' nontranscribed sequence contains all elements required for the efficient heat-controlled expression of the human gene. Two adjacent identical sequence elements, which are partly homologous to the Drosophila "heat shock consensus" sequence, are located 57 to 76 bp upstream from the capping site. Interestingly, the capping site itself is flanked by inverted repeat sequences.
Differential expression of heat shock protein (HSP70) mRNAs in rat cells
Experimental cell …, 1996
Heat shock treatment in PC12 cells induces the synthesis of two HSP70 transcripts of 2.55 and 3.05 kb in size, while only a 70-kDa protein is shown by SDS???PAGE. Using reverse transcription???polymerase chain reaction strategy, two mRNA species were amplified from ...
International Journal of Sciences: Basic and Applied Research, 2018
Heat shock response is rapidly induced to protect cells from irreversible injury by stabilizing cellular synthetic and metabolic activities. Particularly, the response differentially protects against stresses, infections and inflammations. While it could be time- and tissue-dependent, it is not established as to how it exhibits. This study explored the expression of heat shock protein (Hsp) 27, 70 and 90 in intestines, kidneys, livers and lungs of mice undergoing hyperthermia at 42 ° C for 1 hr and recovering for 1-72 hr. The expression of these Hsps was determined by immunohistochemical staining of formalin-fixed paraffin-embedded tissue sections. There was an increase in the expression of Hsp27, 70 and 90 that peaked at 6-12 hr and differentially declined at 48-72 hr. The peak expression was reached earlier in the lung and dropped sharply in the intestine while sustained for a longer time in the liver and kidney. At 72 hr only Hsp90 exhibited moderate expression in the lung and ki...
Heat shock proteins: functions and role in adaptation to hyperthermia
Russian Journal of …, 2005
The results are generalized of many-year studies into the adaptive role of heat shock proteins in different animals, including the representatives of cold-and warm-blooded species that inhabit regions with different thermal conditions. Adaptive evolution of the response to hyperthermia can lead to different results depending on the species. The thermal threshold of induction of the heat shock proteins in desert thermophylic species is, as a rule, higher than in the moderate climate species. In addition, thermoresistant species are often characterized by a certain level of heat shock proteins in cells even at a physiologically normal temperature. Although adaptation to hyperthermia is achieved in most cases without changes in the number of heat shock genes, they can be amplified in some cases in termophylic species. The role of mobile elements in evolution of the heat shock genes was shown and approach was developed for directional introduction of mutations in the promoter regions of these genes.
Molecular and cellular biology, 1993
Transcriptional activity of heat shock (hsp) genes is controlled by a heat-activated, group-specific transcription factor(s) recognizing arrays of inverted repeats of the element NGAAN. To date genes for two human factors, HSF1 and HSF2, have been isolated. To define their properties as well as the changes they undergo during heat stress activation, we prepared polyclonal antibodies to these factors. Using these tools, we have shown that human HeLa cells constitutively synthesize HSF1, but we were unable to detect HSF2. In unstressed cells HSF1 is present mainly in complexes with an apparent molecular mass of about 200 kDa, unable to bind to DNA. Heat treatment induces a shift in the apparent molecular mass of HSF1 to about 700 kDa, concomitant with the acquisition of DNA-binding ability. Cross-linking experiments suggest that this change in complex size may reflect the trimerization of monomeric HSF1. Human HSF1 expressed in Xenopus oocytes does not bind DNA, but derepression of DN...
Induction of a subgroup of acute phase protein genes in mouse liver by hyperthermia
Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1998
We have demonstrated that two members of the acute phase reactant family of positively regulated genes, a -acid 1 Ž . Ž . glycoprotein AGP-1 and AGP-2 and C-reactive protein CRP are induced by hyperthermia, while two others, the serum Ž . Ž . Ž . amyloid A SAA and a -antitrypsin AT genes, are not. Albumin ALB , a negative acute phase reactant gene, is also 1 induced by hyperthermia. The AGP-1, AGP-2, and CRP genes require glucocorticoids, but not IL-6, IL-1b or TNFa in response to hyperthermia. As with LPS, the CrEBPb mRNA levels increased, while the CrEBPa mRNA levels decreased in response to LPS. In contrast to the LPS response, CrEBPd was unchanged. Protein pool levels and DNA-binding activities of the 35 and 20 kDa CrEBPb isoforms increase, whereas protein pool levels of the 42 kDa CrEBPa decrease and the 30 kDa remained high. These studies suggest that the synthesis of specific CrEBPa and CrEBPb isoforms is induced by hyperthermia, and that the regulation of the AGP-1 and AGP-2 genes during heat stress may involve one of these isoforms. The difference between the responses to hyperthermia and LPS is that the former, may not involve the Ž . participation of cytokines. Furthermore, since cis-acting heat shock elements HSE are located in the promoter regions of the ALB, CRP, and CrEBPb genes, these regulatory sequences may be involved in the in vivo activation of these genes by hyperthermia. q 1998 Elsevier Science B.V.
Thermotolerance and heat shock protein induction by slow rates of heating
International Journal of Radiation Oncology Biology Physics, 1988
The magnitude of thermotolerance and the level of heat shock protein (HSP) expression have heen measured in Chinese hamster ovary cells after gradual temperature transients from 37" or 39" to 42" or 43°C. When the level of thermotolerance was measured by clonogenic survival after challenging temperatures between 42" and 43", substantial thermotolerance was observed. However, when the challenging temperature was raised to 45"C, proportionally less thermotolerance was apparent. Heat shock proteins were quantitated by scanning densitometry of radiographs and, in the case of HSP 70, by immunoassay. Scanning densitometry revealed that low levels of heat shock proteins were synthesized during the heating gradients, but less than after a heat shock at 45°C that delivered an equivalent heat dose. The immunoassay of HSP 70 levels measures both pre-existing and newly synthesized protein, and showed that there was no net increase in HSP 70 during two of the heating gradients tested, despite the increase in synthesis noted on the gels. Higher turnover of HSP 70 at the elevated temperatures possibly accounted for the failure to detect a net gain in total protein. In contrast, the total amount of HSP 70 doubled during the 6 hr following a heat shock of 45" for 10 min, an equivalent heat dose to one of the gradients where no net increase in HSP 70 was measured by immunoassay. It appears, then, that tolerance to hyperthemia at 43°C or below may occur under some conditions in the absence of elevated levels of HSP 70, but tolerance to higher temperatures is more closely correlated with increased levels of heat shock proteins. However, even at higher temperatures, our data show disparities between the levels of HSP measured and the thermotolerance expressed. Slow heating, Thermotolerance, Heat shock proteins, Chinese hamster ovary cells.