scholarly journals Skeletal muscle interleukin-6 regulation in hyperthermia

2013 ◽  
Vol 305 (4) ◽  
pp. C406-C413 ◽  
Author(s):  
Steven S. Welc ◽  
Andrew R. Judge ◽  
Thomas L. Clanton
Keyword(s):  
Heat Shock ◽  
Promoter Activity ◽  
Transcriptional Control ◽  
Dominant Negative ◽  
Luciferase Reporter ◽  
C2c12 Myotubes ◽  
Heat Shock Factor 1 ◽  
Activator Protein 1 ◽  
Regulatory Pathways ◽  
Luciferase Reporter Plasmid

We previously reported that IL-6 production is acutely elevated in skeletal muscles exposed to ≥41°C, but the regulatory pathways are poorly understood. The present study characterizes the heat-induced transcriptional control of IL-6 in C2C12 muscle fibers. Hyperthermia exposure (42°C for 1 h) induced transcription from an IL-6 promoter-luciferase reporter plasmid. Heat shock factor-1 (HSF-1), a principal mediator of the heat shock response, was then tested for its role in IL-6 regulation. Overexpression of a constitutively active HSF-1 construct increased basal (37°C) promoter activity, whereas overexpression of a dominant negative HSF-1 reduced IL-6 promoter activity during basal and hyperthermia conditions. Since hyperthermia also induces stress-activated protein kinase (SAPK) signaling, we tested whether mutation of a transcription site downstream of SAPK, (i.e., activator protein-1, AP-1) influences IL-6 transcription in hyperthermia. The mutation had no effect on baseline reporter activity but completely inhibited heat-induced activity. We then tested whether pharmacologically induced states of protein stress, characteristic of cellular responses to hyperthermia and known to induce SAPKs and HSF-1, would induce IL-6 production in the absence of heat. The proteasome was inhibited with MG-132 in one set of experiments, and the unfolded protein response was stimulated with dithiothreitol, thapsigargin, tunicamycin, or castanospermine in other experiments. All treatments stimulated IL-6 protein secretion in the absence of hyperthermia. These studies demonstrate that IL-6 regulation in hyperthermia is directly controlled by HSF-1 and AP-1 signaling and that the IL-6 response in C2C12 myotubes is sensitive to categories of protein stress that reflect accumulation of damaged or unfolded proteins.

Transcriptional Control of Parturition: Insights from Gene Regulation Studies in the Myometrium

2021 ◽  
Author(s):  
Nawrah Khader ◽  
Virlana M Shchuka ◽  
Oksana Shynlova ◽  
Jennifer A Mitchell
Keyword(s):  
Transcription Factors ◽  
Regulatory Networks ◽  
Transcriptional Control ◽  
Progesterone Receptors ◽  
Activator Protein 1 ◽  
Transcriptional Regulatory Networks ◽  
Regulatory Pathways ◽  
Transcriptional Regulatory ◽  
Mechanistic Basis ◽  
Labour Onset

Abstract The onset of labour is a culmination of a series of highly coordinated and preparatory physiological events that take place throughout the gestational period. In order to produce the associated contractions needed for fetal delivery, smooth muscle cells in the muscular layer of the uterus (i.e. myometrium) undergo a transition from quiescent to contractile phenotypes. Here, we present the current understanding of the roles transcription factors play in critical labour-associated gene expression changes as part of the molecular mechanistic basis for this transition. Consideration is given to both transcription factors that have been well-studied in a myometrial context, i.e. activator protein 1 (AP-1), progesterone receptors (PRs), estrogen receptors (ERs), and nuclear factor kappa B (NF-κB), as well as additional transcription factors whose gestational event-driving contributions have been demonstrated more recently. These transcription factors may form pregnancy- and labour- associated transcriptional regulatory networks in the myometrium to modulate the timing of labour onset. A more thorough understanding of the transcription factor-mediated, labour-promoting regulatory pathways holds promise for the development of new therapeutic treatments that can be used for the prevention of preterm labour in at-risk women.

scholarly journals c-Myc cooperates with activated Ras to induce the cdc2 promoter.

1994 ◽  
Vol 14 (9) ◽  
pp. 5710-5718 ◽  
Author(s):  
T L Born ◽  
J A Frost ◽  
A Schönthal ◽  
G C Prendergast ◽  
J R Feramisco
Keyword(s):  
Cell Cycle ◽  
Promoter Activity ◽  
Cell Cycle Progression ◽  
Leucine Zipper ◽  
Luciferase Reporter ◽  
Distinct Region ◽  
Dimerization Domain ◽  
Ras Gene ◽  
Luciferase Reporter Plasmid ◽  
Constitutively Active Mutants

Expression of c-myc with constitutively active mutants of the ras gene results in the cooperative transformation of primary fibroblasts, although the precise mechanism by which these genes cooperate is unknown. Since c-Myc has been shown to function as a transcriptional activator, we have examined the ability of c-Myc and activated Ras (H-RasV-12) to cooperatively induce the promoter activity of cdc2, a gene which is critical for cell cycle progression. Microinjection of expression constructs encoding H-RasV-12 and c-Myc along with a cdc2 promoter-luciferase reporter plasmid into quiescent cells led to an increase in cdc2 promoter activity approximately 30 h after injection, a period which coincides with the S-to-G2/M transition in these cells. Expression of H-RasV-12 alone weakly activated the cdc2 promoter, while expression of c-Myc alone had no effect. Mutants of c-Myc lacking either the leucine zipper dimerization domain or the phosphoacceptor site Ser-62 could not cooperate with H-RasV-12 to induce the cdc2 promoter. These mutants also lacked the ability to cooperate with H-RasV-12 to stimulate DNA synthesis. Deletion analysis identified a distinct region of the cdc2 promoter which was required for c-Myc responsiveness. Taken together, these observations suggest a mechanistic link between the molecular activities of c-Myc and Ras and induction of the cell cycle regulator Cdc2.

scholarly journals Twelve hours of heat stress induces inflammatory signaling in porcine skeletal muscle

2016 ◽  
Vol 310 (11) ◽  
pp. R1288-R1296 ◽  
Author(s):  
Shanthi Ganesan ◽  
Carmen Reynolds ◽  
Katrin Hollinger ◽  
Sarah C. Pearce ◽  
Nicholas K. Gabler ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Heat Stress ◽  
Heat Shock ◽  
Relative Abundance ◽  
Protein Abundance ◽  
Heat Shock Factor 1 ◽  
Activator Protein 1 ◽  
Inflammatory Signaling ◽  
Skeletal Muscle Dysfunction ◽  
Livestock Productivity

Heat stress causes morbidity and mortality in humans and animals and threatens food security by limiting livestock productivity. Inflammatory signaling may contribute to heat stress-mediated skeletal muscle dysfunction. Previously, we discovered increased circulating endotoxin and intramuscular oxidative stress and TNF-α protein abundance, but not inflammatory signaling following 24 and 72 h of heat stress. Thus the purpose of this investigation was to clarify the role of inflammatory signaling in heat-stressed skeletal muscle. Crossbred gilts ( n = 8/group) were assigned to either thermal neutral (24°C), heat stress (37°C), or pair-fed thermal neutral (24°C) conditions for 12 h. Following treatment, animals were euthanized, and the semitendinosus red (STR) and white (STW) were recovered. Heat stress did not alter inflammatory signaling in STW. In STR, relative heat shock protein abundance was similar between groups, as was nuclear content of heat shock factor 1. In whole homogenate, relative abundance of the NF-κB activator inhibitory κB kinase-α was increased by heat stress, although abundance of NF-κB was similar between groups. Relative abundance of phosphorylated NF-κB was increased by heat stress in nuclear fractions. Activator protein-1 (AP-1) signaling was similar between groups. While there were few differences in transcript expression between thermal neutral and heat stress, 80 and 56% of measured transcripts driven by NF-κB or AP-1, respectively, were increased by heat stress compared with pair-fed thermal neutral. Heat stress also caused a reduction in IL-6 transcript and relative protein abundance. These data demonstrate that short-term heat stress causes inflammatory signaling through NF-κB in oxidative, but not glycolytic, skeletal muscle.

scholarly journals Sp1-Mediated Transcription of the Werner Helicase Gene Is Modulated by Rb and p53

1998 ◽  
Vol 18 (11) ◽  
pp. 6191-6200 ◽  
Author(s):  
Yukako Yamabe ◽  
Akira Shimamoto ◽  
Makoto Goto ◽  
Jun Yokota ◽  
Minoru Sugawara ◽  
...  
Keyword(s):  
Transcription Initiation ◽  
Promoter Activity ◽  
Transcriptional Control ◽  
Housekeeping Genes ◽  
Regulatory Elements ◽  
Initiation Site ◽  
Luciferase Reporter ◽  
Upstream Region ◽  
Control Element ◽  
Mrna Levels

ABSTRACT The regulation of Werner’s syndrome gene (WRN) expression was studied by characterizing the cis-regulatory elements in the promoter region and the trans-activating factors that bind to them. First, we defined the transcription initiation sites and the sequence of the 5′ upstream region (2.8 kb) ofWRN that contains a number of cis-regulatory elements, including 7 Sp1, 9 retinoblastoma control element (RCE), and 14 AP2 motifs. A region consisting of nucleotides −67 to +160 was identified as the principal promoter of WRN by reporter gene assays in HeLa cells, using a series of WRNpromoter-luciferase reporter (WRN-Luc) plasmids that contained the 5′-truncated or mutated WRN upstream regions. In particular, two Sp1 elements proximal to the transcription initiation site are indispensable for WRN promoter activity and bind specifically to Sp1 proteins. The RCE enhances WRN promoter activity. Coexpression of the WRN-Luc plasmids with various dosages of plasmids expressing Rb or p53 in Saos2 cells lacking active Rb and p53 proteins showed that the introduced Rb upregulates WRN promoter activity a maximum of 2.5-fold, while p53 downregulates it a maximum of 7-fold, both dose dependently. Consistently, the overexpressed Rb and p53 proteins also affected the endogenous WRN mRNA levels in Saos2 cells, resulting in an increase with Rb and a decrease with p53. These findings suggest that WRN expression, like that of other housekeeping genes, is directed mainly by the Sp1 transcriptional control system but is also further modulated by transcription factors, including Rb and p53, that are implicated in the cell cycle, cell senescence, and genomic instability.

scholarly journals Expression of a Dominant Negative Heat Shock Factor-1 Construct Inhibits Aneuploidy in Prostate Carcinoma Cells*

2004 ◽  
Vol 279 (31) ◽  
pp. 32651-32659 ◽  
Author(s):  
Yiqun Wang ◽  
Jimmy R. Theriault ◽  
Haiying He ◽  
Jianlin Gong ◽  
Stuart K. Calderwood
Keyword(s):  
Heat Shock ◽  
Prostate Carcinoma ◽  
Heat Shock Factor ◽  
Carcinoma Cells ◽  
Dominant Negative ◽  
Heat Shock Factor 1

scholarly journals Heat shock factor‐1 regulates IL‐6 promoter activity in C2C12 muscle myotubes

2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Steven S Welc ◽  
Daniel L Chen ◽  
Andrew R Judge ◽  
Thomas L Clanton
Keyword(s):  
Heat Shock ◽  
Promoter Activity ◽  
Heat Shock Factor ◽  
Heat Shock Factor 1

scholarly journals Isoproterenol and cAMP regulation of the human brain natriuretic peptide gene involves Src and Rac

2000 ◽  
Vol 278 (6) ◽  
pp. E1115-E1123 ◽  
Author(s):  
Quan He ◽  
Guiyun Wu ◽  
Margot C. Lapointe
Keyword(s):  
Brain Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Promoter Activity ◽  
Kinase Inhibitor ◽  
Regulatory Elements ◽  
Dominant Negative ◽  
Luciferase Reporter ◽  
Dominant Negative Mutant ◽  
Luciferase Reporter Gene ◽  
Src Tyrosine Kinase

Brain natriuretic peptide (BNP) gene expression and chronic activation of the sympathetic nervous system are characteristics of the development of heart failure. We studied the role of the β-adrenergic signaling pathway in regulation of the human BNP (hBNP) promoter. An hBNP promoter (−1818 to +100) coupled to a luciferase reporter gene was transferred into neonatal cardiac myocytes, and luciferase activity was measured as an index of promoter activity. Isoproterenol (ISO), forskolin, and cAMP stimulated the promoter, and the β2-antagonist ICI 118,551 abrogated the effect of ISO. In contrast, the protein kinase A (PKA) inhibitor H-89 failed to block the action of cAMP and ISO. Pertussis toxin (PT), which inactivates Gαi, inhibited ISO- and cAMP-stimulated hBNP promoter activity. The Src tyrosine kinase inhibitor PP1 and a dominant-negative mutant of the small G protein Rac also abolished the effect of ISO and cAMP. Finally, we studied the involvement of M-CAT-like binding sites in basal and inducible regulation of the hBNP promoter. Mutation of these elements decreased basal and cAMP-induced activity. These data suggest that β-adrenergic regulation of hBNP is PKA independent, involves a Gαi-activated pathway, and targets regulatory elements in the proximal BNP promoter.

HSF1 alleviates microthrombosis and multiple organ dysfunction in mice with sepsis

2020 ◽  
Author(s):  
Tao Li ◽  
Huan Chen ◽  
Xueyan Shi ◽  
Leijing Yin ◽  
Chuyi Tan ◽  
...  
Keyword(s):  
Heat Shock ◽  
Organ Dysfunction ◽  
Knockout Mice ◽  
Multiple Organ ◽  
Tissue Type ◽  
Luciferase Reporter ◽  
Transcriptional Level ◽  
Heat Shock Factor 1 ◽  
Multiple Organ Dysfunction ◽  
Coagulation Activity

Sepsis is a life-threatening complication of infection and is closely associated with coagulation abnormalities. Heat shock factor 1 (HSF1) is an important transcription factor involved in heat shock response and other biological processes. However, whether HSF1 plays regulatory roles in blood coagulation is still unclear. In this study, a sepsis model was generated in HSF1-knockout mice using caecal ligation and puncture, and the role of HSF1 in microthrombosis and multiple organ dysfunction was evaluated. Notably, lung, liver, and kidney tissues were significantly damaged, fibrin/fibrinogen deposition in the lungs and kidneys was increased, and coagulation activity was gradually increased over time in mice with sepsis; these changes were more obvious in HSF-/- mice than in HSF1+/+ mice. RNA-seq analysis of lung tissues showed that tissue-type plasminogen activator (t-PA) was upregulated in septic mice and was significantly lower in HSF1-knockout mice than in wild-type mice. The effects of HSF1 on t-PA expression were further validated in HSF1-knockout mice with sepsis and in bEnd.3 mouse brain microvascular endothelial cells in vitro using HSF1 RNA interference or overexpression under lipopolysaccharide stimulation. Bioinformatics analysis of the t-PA promoter sequence, combined with electromobility shift and luciferase reporter assays, showed that HSF1 directly up-regulated t-PA at the transcriptional level. Therefore, our results revealed, for the first time, that HSF1 suppressed coagulation activity and microthrombosis by directly up-regulating t-PA, thereby playing a protective role against multiple organ dysfunction in sepsis.

scholarly journals Heat Shock Factor 1 Is a Substrate for p38 Mitogen-Activated Protein Kinases

2016 ◽  
Vol 36 (18) ◽  
pp. 2403-2417 ◽  
Author(s):  
Sharadha Dayalan Naidu ◽  
Calum Sutherland ◽  
Ying Zhang ◽  
Ana Risco ◽  
Laureano de la Vega ◽  
...  
Keyword(s):  
Heat Shock ◽  
P38 Mapk ◽  
Nuclear Translocation ◽  
Heat Shock Factor ◽  
Negative Regulator ◽  
Mitogen Activated Protein Kinase ◽  
Luciferase Reporter ◽  
Heat Shock Factor 1 ◽  
Mitogen Activated Protein ◽  
P38 Mapks

Heat shock factor 1 (HSF1) monitors the structural integrity of the proteome. Phosphorylation at S326 is a hallmark for HSF1 activation, but the identity of the kinase(s) phosphorylating this site has remained elusive. We show here that the dietary agent phenethyl isothiocyanate (PEITC) inhibits heat shock protein 90 (Hsp90), the main negative regulator of HSF1; activates p38 mitogen-activated protein kinase (MAPK); and increases S326 phosphorylation, trimerization, and nuclear translocation of HSF1, and the transcription of a luciferase reporter, as well as the endogenous prototypic HSF1 target Hsp70.In vitro, all members of the p38 MAPK family rapidly and stoichiometrically catalyze the S326 phosphorylation. The use of stable knockdown cell lines and inhibitors indicated that among the p38 MAPKs, p38γ is the principal isoform responsible for the phosphorylation of HSF1 at S326 in cells. A protease-mass spectrometry approach confirmed S326 phosphorylation and unexpectedly revealed that p38 MAPK also catalyzes the phosphorylation of HSF1 at S303/307, previously known repressive posttranslational modifications. Thus, we have identified p38 MAPKs as highly efficient catalysts for the phosphorylation of HSF1. Furthermore, our findings suggest that the magnitude and persistence of activation of p38 MAPK are important determinants of the extent and duration of the heat shock response.

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