Glutamine's protection against cellular injury is dependent on heat shock factor-1

2006 ◽  
Vol 290 (6) ◽  
pp. C1625-C1632 ◽  
Author(s):  
Angela L. Morrison ◽  
Martin Dinges ◽  
Kristen D. Singleton ◽  
Kelli Odoms ◽  
Hector R. Wong ◽  
...  
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Heat Shock Factor ◽  
Tetrazolium Salt ◽  
Heat Shock Factor 1 ◽  
Dependent Manner ◽  
Hsp 70 ◽  
Stress Injury ◽  
Cellular Protection ◽  
Dose Dependent

Glutamine (GLN) has been shown to protect cells, tissues, and whole organisms from stress and injury. Enhanced expression of heat shock protein (HSP) has been hypothesized to be responsible for this protection. To date, there are no clear mechanistic data confirming this relationship. This study tested the hypothesis that GLN-mediated activation of the HSP pathway via heat shock factor-1 (HSF-1) is responsible for cellular protection. Wild-type HSF-1 (HSF-1+/+) and knockout (HSF-1−/−) mouse fibroblasts were used in all experiments. Cells were treated with GLN concentrations ranging from 0 to 16 mM and exposed to heat stress injury in a concurrent treatment model. Cell viability was assayed with phenazine methosulfate plus tetrazolium salt, HSP-70, HSP-25, and nuclear HSF-1 expression via Western blot analysis, and HSF-1/heat shock element (HSE) binding via EMSA. GLN significantly attenuated heat-stress induced cell death in HSF-1+/+ cells in a dose-dependent manner; however, the survival benefit of GLN was lost in HSF-1−/− cells. GLN led to a dose-dependent increase in HSP-70 and HSP-25 expression after heat stress. No inducible HSP expression was observed in HSF-1−/− cells. GLN increased unphosphorylated HSF-1 in the nucleus before heat stress. This was accompanied by a GLN-mediated increase in HSF-1/HSE binding and nuclear content of phosphorylated HSF-1 after heat stress. This is the first demonstration that GLN-mediated cellular protection after heat-stress injury is related to HSF-1 expression and cellular capacity to activate an HSP response. Furthermore, the mechanism of GLN-mediated protection against injury appears to involve an increase in nuclear HSF-1 content before stress and increased HSF-1 promoter binding and phosphorylation.

GLUTAMINE ENHANCES HEAT SHOCK FACTOR-1 NUCLEAR TRANSLOCATION, PHOSHORYLATION AND PROMOTER BINDING FOLLOWING HEAT STRESS INJURY

Shock ◽  
2006 ◽  
Vol 25 (Supplement 1) ◽  
pp. 63
Author(s):  
P. E. Wischmeyer ◽  
A. Morrison ◽  
K. Singleton ◽  
K. Odoms ◽  
H. R. Wong
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Nuclear Translocation ◽  
Heat Shock Factor ◽  
Heat Shock Factor 1 ◽  
Stress Injury

Interaction of Plasminogen and Angiostatin with Heat Shock Proteins.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1622-1622
Author(s):  
Anil K. Dudani ◽  
Jelica Mehic ◽  
Anthony Martyres
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Dependent Manner ◽  
Hsp 70 ◽  
Molar Excess ◽  
Hsp 27 ◽  
Human Plasminogen ◽  
Shock Proteins ◽  
Dose Dependent ◽  
Mcf 7

Abstract Previous studies from this laboratory have demonstrated that plasminogen and angiostatin bind to endothelial cell (EC) surface-associated actin via their kringles in a specific manner. Heat shock proteins (hsps) like hsp 27 are constitutively expressed by vascular ECs and regulate actin polymerization, cell growth and migration. Since many hsps have also been found to be highly abundant on cell surfaces and there is evidence that bacterial surface hsps may interact with human plasminogen, the purpose of this study was to determine whether human plasminogen and angiostatin would interact with human hsps. ELISAs were developed in our laboratory to assess these interactions. It was observed that plasminogen bound to hsps 27, 60 and 70. In all cases, binding was inhibited (85–90%) by excess (50 mM) lysine indicating kringle involvement. Angiostatin predominantly bound to hsp 27 and to hsp 70 in a concentration- and kringle-dependent manner. As observed previously for actin, there was dose-dependent inhibition of angiostatin’s interaction with hsp 27 by plasminogen. In addition, thirty-fold molar excess actin inhibited (up to 50%), the interaction of plasminogen with all hsps. However, thirty-fold molar excess actin could only inhibit the interaction of angiostatin with hsp 27 by 15–20%. FACS analyses indicated the presence of hsps 27, 60 and 70 on the surface of MCF-7 breast cancer cells but not on human umbilical vein ECs. Polyclonal antibodies to hsp 27 significantly inhibited the interaction of plasminogen and angiostatin with MCF-7 surface-associated hsp27 in a dose-dependent manner. Collectively, these data indicate that while plasminogen interacts specifically with hsp 27, 60 and 70, angiostatin interacts predominantly with hsp 27 and to some extent with hsp 70; plasminogen only partially displaces angiostatins binding to hsp 27; actin only partially displaces plasminogen/angiostatin binding to hsps and surface-associated hsp 27 can mediate the binding of both plasminogen and angiostatin to MCF-7 cells.

scholarly journals Molecular Stress-inducing Compounds Increase Osteoclast Formation in a Heat Shock Factor 1 Protein-dependent Manner

2014 ◽  
Vol 289 (19) ◽  
pp. 13602-13614 ◽  
Author(s):  
Ryan C. Chai ◽  
Michelle M. Kouspou ◽  
Benjamin J. Lang ◽  
Chau H. Nguyen ◽  
A. Gabrielle J. van der Kraan ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Factor ◽  
Osteoclast Formation ◽  
Heat Shock Factor 1 ◽  
Dependent Manner

Human heat shock factor 1 is predominantly a nuclear protein before and after heat stress

1999 ◽  
Vol 112 (16) ◽  
pp. 2765-2774 ◽  
Author(s):  
P.A. Mercier ◽  
N.A. Winegarden ◽  
J.T. Westwood
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Nuclear Protein ◽  
Heat Shock Factor ◽  
Nuclear Fraction ◽  
Heat Shock Factor 1 ◽  
Heat Shock Genes ◽  
Before And After ◽  
Biochemical Fractionation ◽  
Multistep Process

The induction of the heat shock genes in eukaryotes by heat and other forms of stress is mediated by a transcription factor known as heat shock factor 1 (HSF1). HSF1 is present in unstressed metazoan cells as a monomer with low affinity for DNA, and upon exposure to stress it is converted to an ‘active’ homotrimer that binds the promoters of heat shock genes with high affinity and induces their transcription. The conversion of HSF1 to its active form is hypothesized to be a multistep process involving physical changes in the HSF1 molecule and the possible translocation of HSF1 from the cytoplasm to the nucleus. While all studies to date have found active HSF1 to be a nuclear protein, there have been conflicting reports on whether the inactive form of HSF is predominantly a cytoplasmic or nuclear protein. In this study, we have made antibodies against human HSF1 and have reexamined its localization in unstressed and heat-shocked human HeLa and A549 cells, and in green monkey Vero cells. Biochemical fractionation of heat-shocked HeLa cells followed by western blot analysis showed that HSF1 was mostly found in the nuclear fraction. In extracts made from unshocked cells, HSF1 was predominantly found in the cytoplasmic fraction using one fractionation procedure, but was distributed approximately equally between the cytoplasmic and nuclear fractions when a different procedure was used. Immunofluorescence microscopy revealed that HSF1 was predominantly a nuclear protein in both heat shocked and unstressed cells. Quantification of HSF1 staining showed that approximately 80% of HSF1 was present in the nucleus both before and after heat stress. These results suggest that HSF1 is predominantly a nuclear protein prior to being exposed to stress, but has low affinity for the nucleus and is easily extracted using most biochemical fractionation procedures. These results also imply that HSF1 translocation is probably not part of the multistep process in HSF1 activation for many cell types.

scholarly journals Neurospora crassa heat shock factor 1 Is an Essential Gene; a Second Heat Shock Factor-Like Gene, hsf2, Is Required for Asexual Spore Formation

2008 ◽  
Vol 7 (9) ◽  
pp. 1573-1581 ◽  
Author(s):  
Seona Thompson ◽  
Nirvana J. Croft ◽  
Antonis Sotiriou ◽  
Hugh D. Piggins ◽  
Susan K. Crosthwaite
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Neurospora Crassa ◽  
Essential Gene ◽  
Expression Profiles ◽  
Heat Shock Factor ◽  
Model Organisms ◽  
Heat Shock Factor 1 ◽  
Wild Type ◽  
Altered Expression

ABSTRACT Appropriate responses of organisms to heat stress are essential for their survival. In eukaryotes, adaptation to high temperatures is mediated by heat shock transcription factors (HSFs). HSFs regulate the expression of heat shock proteins, which function as molecular chaperones assisting in protein folding and stability. In many model organisms a great deal is known about the products of hsf genes. An important exception is the filamentous fungus and model eukaryote Neurospora crassa. Here we show that two Neurospora crassa genes whose protein products share similarity to known HSFs play different biological roles. We report that heat shock factor 1 (hsf1) is an essential gene and that hsf2 is required for asexual development. Conidiation may be blocked in the hsf2 knockout (hsf2 KO ) strain because HSF2 is an integral element of the conidiation pathway or because it affects the availability of protein chaperones. We report that genes expressed during conidiation, for example fluffy, conidiation-10, and repressor of conidiation-1 show wild-type levels of expression in a hsf2 KO strain. However, consistent with the lack of macroconidium development, levels of eas are much reduced. Cultures of the hsf2 KO strain along with two other aconidial strains, the fluffy and aconidial-2 strains, took longer than the wild type to recover from heat shock. Altered expression profiles of hsp90 and a putative hsp90-associated protein in the hsf2 KO strain after exposure to heat shock may in part account for its reduced ability to cope with heat stress.

scholarly journals Heat stress induces epithelial plasticity and cell migration independent of heat shock factor 1

2012 ◽  
Vol 17 (6) ◽  
pp. 765-778 ◽  
Author(s):  
B. J. Lang ◽  
L. Nguyen ◽  
H. C. Nguyen ◽  
J. L. Vieusseux ◽  
R. C. C. Chai ◽  
...  
Keyword(s):  
Cell Migration ◽  
Heat Stress ◽  
Heat Shock ◽  
Heat Shock Factor ◽  
Heat Shock Factor 1 ◽  
Epithelial Plasticity

Heat-shock factor 1 both positively and negatively affects cellular clonogenic growth depending on p53 status

2013 ◽  
Vol 452 (2) ◽  
pp. 321-329 ◽  
Author(s):  
Chau H. Nguyen ◽  
Benjamin J. Lang ◽  
Ryan C. C. Chai ◽  
Jessica L. Vieusseux ◽  
Michelle M. Kouspou ◽  
...  
Keyword(s):  
Heat Shock ◽  
Cancer Cell ◽  
Heat Shock Factor ◽  
Mutant P53 ◽  
Heat Shock Factor 1 ◽  
Dependent Manner ◽  
Wild Type ◽  
Clonogenic Growth ◽  
Wild Type P53

HSF1 (heat-shock factor 1) is the master regulator of the heat-shock response; however, it is also activated by cancer-associated stresses and supports cellular transformation and cancer progression. We examined the role of HSF1 in relation to cancer cell clonogenicity, an important attribute of cancer cells. Ectopic expression or HSF1 knockdown demonstrated that HSF1 positively regulated cancer cell clonogenic growth. Furthermore, knockdown of mutant p53 indicated that HSF1 actions were mediated via a mutant p53-dependent mechanism. To examine this relationship more specifically, we ectopically co-expressed mutant p53R273H and HSF1 in the human mammary epithelial cell line MCF10A. Surprisingly, within this cellular context, HSF1 inhibited clonogenicity. However, upon specific knockdown of endogenous wild-type p53, leaving mutant p53R273H expression intact, HSF1 was observed to greatly enhance clonogenic growth of the cells, indicating that HSF1 suppressed clonogenicity via wild-type p53. To confirm this we ectopically expressed HSF1 in non-transformed and H-RasV12-transformed MCF10A cells. As expected, HSF1 significantly reduced clonogenicity, altering wild-type p53 target gene expression levels consistent with a role of HSF1 increasing wild-type p53 activity. In support of this finding, knockdown of wild-type p53 negated the inhibitory effects of HSF1 expression. We thus show that HSF1 can affect clonogenic growth in a p53 context-dependent manner, and can act via both mutant and wild-type p53 to bring about divergent effects upon clonogenicity. These findings have important implications for our understanding of HSF1's divergent roles in cancer cell growth and survival as well as its disparate effect on mutant and wild-type p53.

scholarly journals The interactive association between heat shock factor 1 and heat shock proteins in primary myocardial cells subjected to heat stress

2015 ◽  
Vol 37 (1) ◽  
pp. 56-62 ◽  
Author(s):  
SHU TANG ◽  
HONGBO CHEN ◽  
YANFEN CHENG ◽  
MOHAMMAD ABDEL NASIR ◽  
NICOLE KEMPER ◽  
...  
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Heat Shock Factor ◽  
Heat Shock Factor 1 ◽  
Myocardial Cells ◽  
Shock Proteins
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