Possible involvement of heat shock protein 25 in the angiotensin II-induced glomerular mesangial cell contraction via p38 MAP kinase

We investigated the effect of prostaglandin E2 (PGE2) on the induction of heat shock protein 27 (HSP27) and HSP70, and the mechanism behind the induction in osteoblast-like MC3T3-E1 cells. PGE2 time-dependently increased the level of HSP27 without affecting the level of HSP70. PGE2 stimulated the accumulation of HSP27 dose-dependently in the range between 10 nM and 10 microM. PGE2 stimulated the increase in the level of the mRNA for HSP27. Staurosporine and calphostin C, inhibitors of protein kinase C (PKC), suppressed the PGE2-induced HSP27 accumulation. The effect of PGE2 on HSP27 accumulation was reduced in the PKC down-regulated cells. BAPTA/AM, a chelator of intracellular Ca2+, or TMB-8, an inhibitor of intracellular Ca2+ mobilization, reduced the accumulation of HSP27 induced by PGE2. Dibutyryl cAMP had little effect on the basal level of HSP27. PGE2 induced the phosphorylation of both p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase. PD98059 and U-0126, inhibitors of the upstream kinase of p44/p42 MAP kinase, reduced the accumulation of HSP27 induced by PGE2. SB203580, a specific inhibitor of p38 MAP kinase, suppressed the HSP27 accumulation induced by PGE2. U-73122, an inhibitor of phospholipase C, and calphostin C reduced the PGE2-induced phosphorylation of both p44/p42 MAP kinase and p38 MAP kinase. These results indicate that PGE2 stimulates the induction of HSP27 through PKC-dependent activations of both p44/p42 MAP kinase and p38 MAP kinase in osteoblasts.

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Short-chain fatty acid mediated phosphorylation of heat shock protein 25: effects on camptothecin-induced apoptosis

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00304.2005 ◽

2006 ◽

Vol 291 (2) ◽

pp. G178-G188 ◽

Cited By ~ 9

Author(s):

Kuljit Parhar ◽

Kathy A. Baer ◽

Kristy Parker ◽

Mark J. Ropeleski

Keyword(s):

Fatty Acid ◽

Heat Shock ◽

Heat Shock Protein ◽

Map Kinase ◽

Short Chain Fatty Acid ◽

Trichostatin A ◽

P38 Map Kinase ◽

Chain Fatty Acid ◽

Intestinal Epithelial ◽

Induced Apoptosis

Although short-chain fatty acid (SCFA)-induced heat shock protein 25 (Hsp25) is associated with increased cellular resistance to injury, withdrawal of lumenal butyrate in vivo is associated with intestinal epithelial injury and apoptosis. Recognizing that SCFA-dependent posttranslational modification of Hsp25 may involve altered Hsp25 phosphorylation, we hypothesized that butyrate regulates Hsp25 phosphorylation and secondarily affects cellular responses to apoptosis-inducing agents. Intestinal epithelial crypt IEC-18 cells were treated with butyrate, propionate, or the histone deacetylase inhibitor trichostatin A for 6–24 h. Immunolocalization of Hsp25 was examined by confocal laser microscopy. Hsp25 phosphorylation was characterized using two-dimensional isoelectric focusing gel electrophoresis. Hsp25 accumulation in cytoskeletal- and mitochondrial-enriched fractions was examined by immunoblotting. The activation of p38 MAP kinase was determined using phospho-specific antibodies and MAPKAPK 2 kinase assays. The effects of SCFA on apoptosis were studied by ELISA detection of cleaved DNA and using antibodies recognizing cleaved caspase-3. Five-millimolar butyrate induced no significant injury to IEC-18 cells. Hsp25 did not accumulate in Triton X-100-insoluble cytoskeletal fractions with butyrate treatment but did localize to mitochondria in a p38 MAP kinase-dependent manner. Hsp25 phosphorylation was induced by butyrate, propionate, and trichostatin A. Butyrate-mediated changes in Hsp25 phosphorylation coincide with the activation of the p38 MAP kinase and MAPKAPK 2. Butyrate, propionate, and low-dose trichostatin A confer significant protection from camptothecin-induced apoptosis, which was not reversed by the p38 inhibitor SB203580. We conclude that butyrate-mediated phosphorylation of Hsp25 is associated with significant resistance to apoptosis, which appears to be independent of p38-mediated targeting of Hsp25 to mitochondria.

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Phosphorylation of the 27-kDa heat shock protein via p38 MAP kinase and MAPKAP kinase in smooth muscle

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1997.273.5.l930 ◽

1997 ◽

Vol 273 (5) ◽

pp. L930-L940 ◽

Cited By ~ 37

Author(s):

Janice K. Larsen ◽

Ilia A. Yamboliev ◽

Lee A. Weber ◽

William T. Gerthoffer

Keyword(s):

Smooth Muscle ◽

Stress Response ◽

Heat Shock ◽

Heat Shock Protein ◽

Map Kinase ◽

Tyrosine Phosphorylation ◽

Airway Smooth Muscle ◽

Muscarinic Receptors ◽

P38 Map Kinase ◽

Mitogen Activated Protein

The 27-kDa heat shock protein (HSP27) is expressed in a variety of tissues in the absence of stress and is thought to regulate actin filament dynamics, possibly by a phosphorylation/dephosphorylation mechanism. HSP27 has also been suggested to be involved in contraction of intestinal smooth muscle. We have investigated phosphorylation of HSP27 in airway smooth muscle in response to the muscarinic agonist carbachol. Carbachol increased32P incorporation into canine tracheal HSP27 and induced a shift in the distribution of charge isoforms on two-dimensional gels to more acidic, phosphorylated forms. The canine HSP27 amino acid sequence includes three serine residues corresponding to sites in human HSP27 known to be phosphorylated by mitogen-activated protein kinase-activated protein (MAPKAP) kinase-2. To determine whether muscarinic receptors are coupled to a “stress response” pathway in smooth muscle culminating in phosphorylation of HSP27, we assayed MAPKAP kinase-2 activity and tyrosine phosphorylation of p38 mitogen-activated protein (MAP) kinase, the enzyme thought to activate MAPKAP kinase-2. Recombinant canine HSP27 expressed in Escherichia coli was a substrate for MAPKAP kinase-2 in vitro as well as a substrate for endogenous smooth muscle HSP27 kinase, which was activated by carbachol. Carbachol also increased tyrosine phosphorylation of p38 MAP kinase. SB-203580, an inhibitor of p38 MAP kinases, reduced activation of endogenous HSP27 kinase activity and blocked the shift in HSP27 charge isoforms to acidic forms. We suggest that HSP27 in airway smooth muscle, in addition to being a stress response protein, is phosphorylated by a receptor-initiated signaling cascade involving muscarinic receptors, tyrosine phosphorylation of p38 MAP kinase, and activation of MAPKAP kinase-2.

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Modulation of actin dynamics during stress and physiological stimulation by a signaling pathway involving p38 MAP kinase and heat-shock protein 27

Biochemistry and Cell Biology ◽

10.1139/o95-078 ◽

1995 ◽

Vol 73 (9-10) ◽

pp. 703-707 ◽

Cited By ~ 199

Author(s):

Jacques Landry ◽

Jacques Huot

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Actin Cytoskeleton ◽

Map Kinase ◽

Signaling Pathway ◽

Actin Polymerization ◽

P38 Map Kinase ◽

Actin Dynamics ◽

Protective Function ◽

Short Period

HSP27, like other proteins of the heat-shock protein family, accumulates to high levels after exposure of cells to a short period of hyperthermia and contributes to the development of a transient state of thermoresistance. In vitro, HSP27 behaves as an actin cap-binding protein and can inhibit actin polymerization. In vivo, the protective function of HSP27 is exerted mainly at the level of the microfilaments and appears as an extension of a normal function of the protein. This function is regulated by phosphorylation in a mitogen- and stress-sensitive signaling pathway involving the newly characterized p38 MAP kinase. The phosphorylation-modulated function of HSP27 can contribute to agonist-induced reorganization of the actin cytoskeleton and, in the case of stress activation, provides an actin-based adaptive response of cells to the new environmental conditions.Key words: actin cytoskeleton, heat-shock protein, HSP27, signal transduction, stress response, mitogen activated protein kinases.

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Endothelin-1 stimulates heat shock protein 27 induction in osteoblasts: involvement of p38 MAP kinase

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1999.277.6.e1046 ◽

1999 ◽

Vol 277 (6) ◽

pp. E1046-E1054 ◽

Cited By ~ 4

Author(s):

Hidenori Kawamura ◽

Takanobu Otsuka ◽

Hiroyuki Matsuno ◽

Masayuki Niwa ◽

Nobuo Matsui ◽

...

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Map Kinase ◽

Heat Shock Protein 27 ◽

P38 Map Kinase ◽

Mrna Levels ◽

Endothelin 1 ◽

Calphostin C ◽

Hsp 27 ◽

Sb 203580

We previously reported that endothelin-1 (ET-1) activates p42/p44 mitogen-activated protein (MAP) kinase in osteoblast-like MC3T3-E1 cells and consequently induces synthesis of interleukin-6. In the present study, we investigated the effect of ET-1 on the induction of heat shock protein 27 (HSP 27) in MC3T3-E1 cells. ET-1 time and dose dependently stimulated HSP 27 accumulation. ET-1 induced an increase in the levels of mRNA for HSP 27. Both staurosporine and calphostin C, inhibitors of protein kinase C (PKC), suppressed the ET-1-induced HSP 27 accumulation. 12- O-tetradecanoylphorbol 13-acetate (TPA), a PKC activator, induced the HSP 27 accumulation and the expression of mRNA for HSP 27. The ET-1-stimulated HSP 27 accumulation was reduced in PKC-downregulated MC3T3-E1 cells. The HSP 27 accumulation by ET-1 was not suppressed by PD-98059, an inhibitor of the upstream kinase that activates p42/p44 MAP kinase. ET-1 or TPA induced the phosphorylation of p38 MAP kinase. SB-203580, an inhibitor of p38 MAP kinase, reduced the ET-1-stimulated HSP 27 accumulation. Calphostin C and U-73122, a phospholipase C inhibitor, suppressed the ET-1-induced phosphorylation of p38 MAP kinase. U-73122 and propranolol, a phosphatidic acid phosphohydrolase inhibitor, reduced the ET-1-stimulated HSP 27 accumulation. SB-203580 suppressed the ET-1-stimulated increase in the mRNA levels for HSP 27. These results strongly suggest that ET-1 stimulates HSP 27 induction in osteoblasts and that p38 MAP kinase activation is involved in the HSP 27 induction.

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JlpA of Campylobacter jejuni interacts with surface-exposed heat shock protein 90alpha and triggers signalling pathways leading to the activation of NF-kappaB and p38 MAP kinase in epithelial cells

Cellular Microbiology ◽

10.1046/j.1462-5822.2003.00265.x ◽

2003 ◽

Vol 5 (3) ◽

pp. 165-174 ◽

Cited By ~ 88

Author(s):

Songmu Jin ◽

Young C. Song ◽

Andrew Emili ◽

Philip M. Sherman ◽

Voon Loong Chan

Keyword(s):

Heat Shock ◽

Epithelial Cells ◽

Heat Shock Protein ◽

Map Kinase ◽

Campylobacter Jejuni ◽

P38 Map Kinase ◽

Signalling Pathways ◽

Nf Kappab

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Involvement of stress-activated protein kinase/c-Jun N-terminal kinase in endothelin-1-induced heat shock protein 27 in osteoblasts

Acta Endocrinologica ◽

10.1530/eje.0.1490239 ◽

2003 ◽

pp. 239-245 ◽

Cited By ~ 8

Author(s):

H Tokuda ◽

M Niwa ◽

H Ito ◽

Y Oiso ◽

K Kato ◽

...

Keyword(s):

Protein Kinase C ◽

Heat Shock ◽

Protein Kinase ◽

Heat Shock Protein ◽

Map Kinase ◽

Blot Analysis ◽

Heat Shock Protein 27 ◽

P38 Map Kinase ◽

Endothelin 1 ◽

Kinase C

OBJECTIVE: We have reported that endothelin-1 (ET-1) activates p38 mitogen-activated protein (MAP) kinase through protein kinase C in osteoblast-like MC3T3-E1 cells, and that p38 MAP kinase plays a role in the ET-1-induced heat shock protein 27 (HSP27). Recently, we found that stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) is activated by ET-1 in these cells. In the present study, we have investigated the involvement of SAPK/JNK in ET-1-induced HSP27 in MC3T3-E1 cells. METHODS: The concentration of HSP27 in soluble extracts of the cells, the expression of mRNA for HSP27, and the phosphorylation of SAPK/JNK were determined by an enzyme immunoassay, Northern blot analysis, and Western blot analysis respectively. RESULTS: SP600125, a specific inhibitor of SAPK/JNK, markedly reduced ET-1-stimulated HSP27 accumulation. The inhibitory effect of SP600125 was dose dependent in the range between 1 and 50 microM. SP600125 reduced the ET-1-increased level of HSP27 mRNA. Calphostin C and Go 6976, inhibitors of protein kinase C, reduced the ET-1-induced phosphorylation of SAPK/JNK. 12-O-Tetradecanoylphorbol-13-acetate, a direct activator of protein kinase C, induced SAPK/JNK phosphorylation, which was suppressed by SP600125. A combination of SP600125 and p38 MAP kinase inhibitor such as SB203580 and PD169316 additively reduced the ET-1-stimulated accumulation of HSP27. CONCLUSIONS: These results strongly suggest that JNK plays a part in ET-1-induced HSP27 in addition to p38 MAP kinase in osteoblasts.

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