Differential p38-dependent signalling in response to cellular stress and mitogenic stimulation in fibroblasts
© Faust et al; licensee BioMed Central Ltd. 2012
Received: 14 September 2011
Accepted: 9 March 2012
Published: 9 March 2012
p38 MAP kinase is known to be activated by cellular stress finally leading to cell cycle arrest or apoptosis. Furthermore, a tumour suppressor role of p38 MAPK has been proposed. In contrast, a requirement of p38 for proliferation has also been described. To clarify this paradox, we investigated stress- and mitogen-induced p38 signalling in the same cell type using fibroblasts. We demonstrate that - in the same cell line - p38 is activated by mitogens or cellular stress, but p38-dependent signalling is different. Exposure to cellular stress, such as anisomycin, leads to a strong and persistent p38 activation independent of GTPases. As a result, MK2 and downstream the transcription factor CREB are phosphorylated. In contrast, mitogenic stimulation results in a weaker and transient p38 activation, which upstream involves small GTPases and is required for cyclin D1 induction. Consequently, the retinoblastoma protein is phosphorylated and allows G1/S transition. Our data suggest a dual role of p38 and indicate that the level and/or duration of p38 activation determines the cellular response, i.e either proliferation or cell cycle arrest.
Keywordsp38 MAPK Signalling Cellular stress Mitogens Fibroblasts
The family of mitogen-activated protein kinases (MAPKs) involves ERKs (extracellular signal-regulated kinases), JNKs (c-Jun-N-terminal kinases) and p38 MAPKs. They are proline-directed Ser/Thr kinases mediating a variety of cellular responses due to numerous extracellular stimuli. While the ERK pathway is preferentially induced by mitogens, p38 and JNK are generally activated by inflammatory cytokines and cellular stress, such as hyperosmolarity, heat shock, genotoxic compounds, UV light, γ-irradiation, metabolic stress, and protein synthesis inhibition [for review see [1–4]]. Mammalian p38 was first cloned by Han and coworkers and revealed close homology to the yeast osmosensing HOG1 . A common response of p38 activation by cellular stress is cell cycle arrest or apoptosis. Moreover and consistent with its role in cell cycle regulation, p38 is involved in oncogene-induced senescence, replicative senescence, differentiation, and DNA-damage response [for review see ]. For instance, several reports indicate an inhibitory role of p38 in proliferation in the mouse fibroblast cell line NIH3T3. Tunicamycin-treatment causes activation of p38 resulting in phosphorylation of GADD153 finally leading to cell cycle block . G1-arrest is also detected in response to arsenite, which is mediated by a p38-dependent increase in p21 . Constitutive expression of oncogenic ras results in sustained activation of p38 and inhibition of proliferation . Consistent with a negative role in cell cycle progression, expression of active MKK3/6 induces G1 arrest in the same cell line . Similar data has been obtained in rhabdomyosarcoma cells, in which p38 activation results in G1-arrest and differentiation . We recently revealed that sustained activation of p38 in response to high cell density is involved in the signalling cascade of contact-inhibition in murine and in human FH109 fibroblasts by regulating levels of the Cdk-inhibitor p27 .
However, several reports suggest that p38 is also activated in mitogenic pathways. Maher described a requirement of p38 for bFGF-induced (but not for PDGF-stimulated) cell proliferation in Swiss3T3 fibroblasts . A similar role has been proposed in hepatocytes after EGF- and insulin-treatment , and ERK cooperates with p38 in G-CSF-stimulated hematopoietic cell proliferation . A proliferative role of p38 has also been described in breast cancer-, chondrosarcoma-, and melanoma cells [16–18]. These reports suggest a proliferative function of p38 in contrast to the above mentioned role of p38 in stress response and cell cycle arrest.
Some simple explanations for these discrepancies could be dependence on stimulus, cellular context or cell-type specificity. For example, p38 activation by one type of stimulus might lead to different biological outcomes, i.e. proliferation or growth arrest, dependent on cell type. Alternatively, distinct stimuli could induce p38 signalling via different routes in the very same cell type thus triggering diverging responses. Unfortunately, the proliferative and growth inhibitory role of p38 has been investigated so far only in different cell lines and thus the possibility of divergent p38 signalling downstream of different stimuli in simply the same cell line is poorly understood. Therefore we studied p38 signalling in human and murine fibroblast cell lines and compared the signalling cascades up- and downstream of p38 in response to mitogens and cellular stress. Here we show that mitogen-induced proliferation in NIH3T3 and FH109 fibroblasts can be substantially blocked by the compound SB203580, which is a selective inhibitor of p38  again demonstrating that p38 mediates cellular proliferation. Since p38 also regulates cell cycle arrest in the same cell lines as mentioned above, we analysed mitogen- and stress-induced activation of p38 in NIH3T3 and FH109 fibroblasts. We provide evidence for a dual role of p38 in cell cycle control and suggest that the level and/or the duration of p38 activation might determine the cell's decision to proliferate or to induce cell cycle arrest. We show that in NIH3T3 and FH109 fibroblast cell lines mitogenic stimuli lead to a weak and transient phosphorylation of p38, which is absolutely required for G1/S-transition whereas anisomycin induces a strong and sustained activation of p38. We further revealed that the signalling cascades involving p38 activation after serum- and stress-treatment differ. While mitogenic activation of p38 upstream involves small GTPases and downstream leads to cyclin D1 expression, anisomycin-dependent activation of p38 is independent of GTPases and leads to phosphorylation of MK2 and finally CREB. In conclusion, our data provide an example of differentially wired p38 signalling in response to distinct stimuli, which results in specific outputs such as stress-induced cell cycle arrest and mitogen-induced proliferation.
Involvement of p38 in mitogen-induced proliferation
To investigate whether p38 is also required for continuous proliferation, cells were seeded and cultured in 10% FCS up to 72 h in the absence or presence of SB203580. Figure 1C clearly shows that continuous proliferation of NIH3T3 cells is impaired in the presence of SB203580.
Activation of p38 after mitogenic stimuli and cellular stress
Cell cycle proteins are differentially affected by p38 signalling after mitogenic stimuli or cellular stress
Phosphorylation of pRB is mediated by cyclin-dependent kinases which are regulated by cyclins, namely by the Cdk4/cyclin D1 complex and the Cdk2/cyclin E complex. Hence, downregulation of cyclin D1 will lead to impaired phosphorylation of pRB. Indeed, serum-induced expression of cyclin D1 is strongly attenuated in the presence of SB203580 (Figure 3C). Downregulation of cyclin D1 expression by SB203580 was still detected 24 h after mitogenic stimulation (Additional file 4A). Thus, inhibition of p38 does not merely delay cyclin D1 expression, but prevents cyclin D1 induction. Similar findings were obtained in FH109 cells (Additional file 4B).
In contrast, anisomycin and sorbitol treatment led to a complete loss of cyclin D1 expression. However, this effect could not be reversed by SB203580 (Figure 3D). Hence, downregulation of cyclin D1 in response to cellular stress is probably mediated in addition by other stress kinases or pathways in NIH3T3 cells.
Transcription factors are differentially targeted by p38 after mitogenic stimuli or cellular stress
FCS-, but not anisomycin-induced phosphorylation of p38 is dependent on GTPases
In the present work we show that p38 is activated by mitogens and cellular stress in the same cell line, but that the signalling pathways differ. We suggest that p38 plays a dual role in cell cycle control in fibroblasts mediating cell cycle progression or cell cycle arrest depending on the extracellular stimulus. In NIH3T3 cells, p38-dependent cell cycle arrest either due to cellular stress or constitutive activation by overexpression of the kinase itself or an upstream activating kinase has been demonstrated in various publications [7–10]. The fact that in the present study in the same cell line stimulation with serum or growth factors results in phosphorylation and activation of p38, and - vice versa - FCS- and growth factor-induced DNA-synthesis is blocked by the p38-specific inhibitor SB203580 or siRNA-mediated knock-down of p38 clearly indicates that p38 is also required for proliferation and points to a dual role of p38 in cell cycle regulation. Our data suggest that a key element might be the duration and/or amount of activation which then leads to different downstream signalling. While anisomycin-exposure leads to a strong and sustained activation of the p38 and MK2 kinases, thereby increasing the phosphorylation of CREB [27, 28], mitogen-induced activation of p38 is weaker and transient, but is required for cyclin D1 expression. In addition, we demonstrate that p38 is differentially regulated in response to anisomycin and mitogens with respect to the involvement of small GTPases (Figure 6).
Dual regulation of a kinase depending on the extracellular stimulus has been proposed in the neuronal cell line PC12 for ERK [for review see ]. In these cells, EGF leads to a transient activation of ERK with cytoplasmic retention resulting in a proliferative cellular response. In contrast, NGF-induced ERK activation in the same cells is sustained and accompanied by a nuclear translocation causing cell cycle arrest and neuronal differentiation.
To better understand the role of p38 in mitogen-induced proliferation, we studied cell cycle proteins, i.e. phosphorylation of pRB and expression of the cyclins D1 and A. It is generally accepted that during G1-phase, cyclin D1/Cdk4 and downstream cyclin E/Cdk2 phosphorylate pRB which then dissociates from the transcription factor E2F allowing transcription of S-phase specific genes, such as cyclin A, and thereby entry into S-phase . Since the mitogen-induced expression of cyclin D1 was strongly reduced in the presence of SB203580 we conclude that expression of cyclin D1 requires the activity of p38. An expected consequence of a decrease in cyclin D1 is less activity of the cyclin D1/Cdk4 complex and in turn less phosphorylation of pRB. Hence, the observed attenuation of pRB phosphorylation in the presence of SB203580 is very likely due to decreased cyclin D1 expression. Downregulation of cyclin A by SB203580 was observed at a later time point, i.e. 14 h after mitogenic stimulation. According to the kinetics of pRB phosphorylation we assume that this time point correlates with early S-phase. We therefore conclude that the decrease in cyclin A is not directly mediated by p38, but rather a consequence of inhibition of pRB phosphorylation by SB203580.
The mechanism of p38-dependent expression of cyclin D1 in our cell system is not known so far. In melanoma cells, p38-ATF-2-dependent expression of cyclin D1 in response to hepatocyte growth factor/scatter factor has been described . Induction of cyclin D1 by pp60v-srcis also mediated via the p38/JNK-ATF-2/CREB pathway in human breast cancer cells . Since we did not detect p38-dependent phosphorylation of ATF-2 nor CREB, an involvement of ATF-2 or CREB in cyclin D1 expression in our cell system is unlikely. One possible explanation comes from the observation, that ERK1/2 phosphorylation is blocked from 6 h on after FCS-stimulation, very likely as a secondary effect of p38 inhibition. Hence, p38 activity seems to be required for sustained ERK1/2 phosphorylation. In fibroblasts, sustained ERK1/2 activity is required for cyclin D1 expression, especially during mid-G1-phase . The underlying mechanism of this cross-talk between p38 and ERK1/2 remains to be elucidated.
To our surprise, inhibition of p38 function by SB203580 did not only block mitogen-induced G0/G1-S transition, but also attenuated continuous proliferation of NIH3T3 cells. This observation is in contrast to data obtained in BJ primary fibroblasts and WI-38 fibroblasts. In these cells, SB203580 does not alter proliferation in exponentially growing cultures, which show doubling rates comparable to our NIH3T3 [36, 37]. However, cell type specific differences might explain different functions of p38 in NIH3T3 cells. Very recently, it was shown that the transcription factor FoxM1 acts downstream from the Ras-MKK3-p38 pathway in NIH3T3 cells . Importantly, FoxM1 is also known to regulate a number of proliferative genes [38; manuscript in preparation]. Although we have not tested, another explanation for the discrepancies could be p53 function. While BJ and WI-38 fibroblast express wild-type p53 [39, 40], the NIH3T3 cells we used are p53-deficient (unpublished observations).
Our observation of a sustained p38 activation after anisomycin- or sorbitol-treatment is in accordance with other reports showing persistent activation of p38 after cellular stress, e.g. in C3H10T1/2 cells in response to anisomycin or UV  or in several cell lines in response to γ-irradiation, genotoxic compounds or during premature senescence [[42, 43], reviewed in ].
We have also shown that sustained activation of p38 is required for contact-inhibition in murine and human fibroblasts . Several mechanisms explaining p38-dependent cell cycle arrest have been described. For instance, p27KIP1, a well-known inhibitor of Cdk2 and Cdk4, is one important downstream target of p38 upon contact-inhibition [12, 44]. The protein p27KIP1 is also upregulated in response to genotoxic agents and here is supposed to be crucial for maintenance of cell cycle arrest . However, we did not observe accumulation of p27KIP1 in response to anisomycin or sorbitol (unpublished oberservation). It is also known that the Cdk inhibitors p21WAF1/CIP1 and p16INK4a mediate p38-dependent senescence, for instance in response to DNA-damaging agents and reactive oxygen species, which might be related to the role of p38 as a tumour suppressor [42, 45–51]. Very recently, p38-dependent induction of p21 due to sorbitol-treatment has been described in nucleus pulposus intervertebral disc cells . Whether p21WAF1/CIP1 or p16INK4a are upregulated in response to anisomycin or sorbitol in fibroblasts needs to be determined.
Moreover, we observed sustained phosphorylation of CREB. Two kinases are known to phosphorylate this transcription factor: MK2 [27, 28] and MSK1 . Since MK2 was also persistently activated in response to anisomycin, we conclude that MK2 at least partially contributes to phosphorylation of CREB. A possible involvement of MSK1 remains to be elucidated. Interestingly, cell cycle arrest due to sustained activation of the cAMP/CREB-pathway was also detected in prostate carcinoma cells, which were chronically exposed to pituitary adenylate-cyclase-activating polypeptide. In contrast, transient stimulation of cAMP/CREB induces proliferation . Constitutive activation of CREB by bacterial toxins leads to G1-arrest in a murine macrophage cell line by induction of p27 and downregulation of cyclin D1 . In accordance, cholera toxin, a potent inducer of cellular accumulation of cAMP and thereby phosphorylation of CREB, is able to cause G1 arrest by upregulation of p27, p21 and downregulation of cyclin D1 in rat and primary human glioma cells . However, we could not reverse downregulation of cyclin D1 in response to sorbitol-exposure by SB203580 arguing that p38 is not the sole entity responsible for the decrease in cyclin D1. This is in line with the observation that arsenite-induced downregulation of cyclin D1 in NIH3T3 cells cannot be restored by SB203580  and that cyclin D1 decrease can also be mediated by JNK . Hence, the precise function of the p38-MK2-CREB axis in anisomycin- or sorbitol-induced cell cycle arrest remains to be determined.
Phosphorylation of CREB in response to mitogenic stimulation could not be blocked by SB203580, which is in line with previous observations that CREB phosphorylation in response to growth factors is mediated by the ERK pathway .
Interestingly, phosphorylation of ATF-2 was not inhibited by SB203580, although it has been identified to be an excellent substrate for p38 in vitro . This observation is in perfect accordance with the work of Hazzalin and coworkers  and the work by Maher  ruling out involvement of p38 in phosphorylating endogenous ATF-2 in fibroblasts. Since phosphorylation of ATF-2 could be blocked by pharmacological inhibition of JNK, phosphorylation of ATF-2 is very likely dependent on JNK .
Persistent activation after anisomycin is consistent with the supposed mechanism of action: anisomycin inhibits protein synthesis hence blocking transcription of phosphatases. As a result, p38 dephosphorylation does not occur in the presence of anisomycin. Furthermore, dephosphorylation of the upstream acting MKK3/6 is inhibited thereby allowing prolonged activation of p38. A similar mechanism has been described for arsenite-induced JNK activation [58, 59]. To gain more insight into upstream events regulating p38 activity in response to mitogens, we identified the involvement of small GTPases and made use of selective bacterial toxins. Clostridium sordellii Lethal Toxin (LT) inhibits Ras, Rac, and Rap function, Clostridium difficile Toxin B abolishes Rac, Cdc42, and Rho function, and Botulinus C3 exoenzyme, in our study used as C2IN-C3 fusion toxin displaying high cell permeability , selectively inhibits Rho function [29, 30]. The observation, that ERK activation was selectively abolished only in the presence of Lethal Toxin indicates selectivity of the toxins. In control experiments with Botulinus C2 toxin, which ADP-ribosylates actin , we ruled out that the observed inhibitory effects of the toxins occurred unspecifically due to degradation of the actin cytoskeleton (unpublished observation). If anisomycin-induced p38 activation is due to its suppression of phosphatases (see above) it should be independent of Rho proteins. Indeed, blocking the activity of the small GTPases Rho, Rac, and Cdc42 by preincubation with Lethal Toxin or Toxin B, had no effect on anisomycin-induced p38 phosphorylation.
On the contrary, serum-induced p38 activation could be blocked by preincubation with Toxin B and Lethal Toxin. In view of the fact that C2IN-C3 had no effect on serum-induced p38 phosphorylation, the results strongly argue against an involvement of Rho and point to a potential role of Rac and/or Ras and Cdc42. Indeed, in overexpression studies, Rac and Cdc42 have been identified to mediate p38 activation [10, 61, 62]. More detailed analysis is required to identify which of the small GTPases is involved in p38-mediated control of proliferation.
We present a novel hypothetical model for p38 function and propose a dual role of p38 activation in cell cycle control. Cellular stress by anisomycin leads to a sustained phosphorylation and activation of p38. One downstream substrate of p38 is MK2 which phosphorylates CREB. In contrast, after mitogenic stimulation p38 is only transiently phosphorylated, but promotes cyclin D1 expression. The cellular response is proliferation. Differential target activation by p38 downstream of mitogenic and stress signals might be related to the respective strength or duration of p38 activation or, alternatively, to additional cross-talk with parallel pathways - an issue which warrants further investigations.
Materials and methods
FH109 human embryonal lung fibroblasts  and NIH3T3 murine fibroblasts were routinely cultured in Dulbecco's Modified Eagle's Medium (DMEM) (PAA), supplemented with 10% fetal calf serum (FCS) (PAA), 4 mM glutamine, penicillin and streptomycin (each 100 U/ml). For experiments, FH109 or NIH3T3 cells were cultured in DMEM/0.2% FCS for 72 h and then treated with anisomycin (10 μg/ml) (Sigma) or FCS to a final concentration of 10% and harvested at different time points as described in the figure legends. Cells were pretreated with SB203580 (10 μΜ) (Calbiochem), or SP600125 (25 μM) (Enzo Life Sciences), control cells were exposed to 0.1% DMSO, the solvent used for the inhibitors.
Measurement of DNA-synthesis
FH109 or NIH3T3 cells were seeded into microtiter plates at a density of 8 × 103/well or 5 × 103/well, respectively, and cultured for 72 h in DMEM/0.2% FCS. Cells were stimulated for 24 h by the addition of 10% FCS, PDGF- or bFGF (each 50 ng/ml, Cell Signaling). SB203580 was added 1 h before stimulation. DNA-synthesis was measured by [3H]thymidine-incorporation as described .
Determination of cell number
Cells were washed, trypsinised and counted in a hemocytometer.
Total cell extracts were prepared by lysing the cells in hot Laemmli sample buffer and protein concentration was determined according to . Equal amounts of protein (20 - 50 μg protein/lane) were separated by SDS-PAGE (7.5 - 10%) and electroblotted overnight onto Immobilon membrane (Millipore). The membranes were blocked for 1 h with 5% low-fat milk-powder or 5% bovine serum albumin in TBS (50 mM Tris-HCl, pH 7.5, 150 mM NaCl) containing 0.05% Tween 20 and then incubated either for 1.5 h at room temperature with anti-cyclin D1-, anti-cyclin A- (1:1000, Santa Cruz), or overnight at 4°C with anti-phospho-pRb- (1:1000, Cell Signaling), anti-phospho-p38-, anti-CREB, anti-phospho-ATF-2-, anti-phospho-c-Jun-, or anti-phospho-MK2-antibody (1:1000-2000, Cell Signaling) followed by incubation with horseradish-peroxidase-conjugated secondary antibody and ECL-detection according to the manufacturer's instructions. The blots were stripped and reprobed with anti-p38α-, anti-phospho-CREB-, (1:1000, Cell Signaling), anti-ATF-2-, anti-PCNA-, anti-ERK-, or anti-c-Jun-antibody (1:1000, Santa Cruz) or anti-gelsolin-antibody (1:200, Santa Cruz) followed by ECL-detection.
Transfection of siRNA
For transient transfection of p38α or control siRNA, 5 × 103 cells/well (96 well plate) were seeded and cultured for 24 h to reach 80-90% confluence. Transfection was performed in a total volume of 120 μl containing 8 pmol siRNA and 0.2 μl of Lipofectamine 2000 according to the manufacturer's instructions. After 24 h, medium was changed to DMEM/0.2% FCS and the cells cultured for another 24 h. Cells were then exposed for 24 h to PDGFβ or bFGF (see above) and incorporation of [3H]thymidine was determined. p38α siRNA (directed against murine p38α mRNA sequence [MGI:1346865]): 5'-GGAAUUCAAUGACGUGU AC-3'; control siRNA (directed against mRNA encoding the red fluorescence protein DsRed from the coral Discosoma) has been published previously .
We are indebted to H. Barth, F. Hofmann and K. Aktories for the generous gifts of the bacterial toxins. The work was supported by the grant Di793/1-1 from the Deutsche Forschungsgemeinschaft and by ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework program, Priority 5: 'Food Quality and Safety' (Contract no. 513943) and is part of the M.D. thesis of C.S.
- Kyriakis J, Avruch J: Protein kinase cascades activated by stress and inflammatory cytokines. Bioessays. 1996, 18: 567-577. 10.1002/bies.950180708.View ArticlePubMedGoogle Scholar
- Junttila MR, Li S-P, Westermarck J: Phosphatase-mediated crosstalk between MAPK signaling pathways in the regulation of cell survival. FASEB J. 2008, 22: 954-964.View ArticlePubMedGoogle Scholar
- Herrlich P, Karin M, Weiss C: Supreme EnLIGHTenment: damage recognition and signaling in the mammalian UV response. Mol Cell. 2008, 29: 279-290. 10.1016/j.molcel.2008.01.001.PubMed CentralView ArticlePubMedGoogle Scholar
- Cuadrado A, Nebreda AR: Mechanisms and functions of p38 MAPK signalling. Biochem J. 2010, 429: 403-417. 10.1042/BJ20100323.View ArticlePubMedGoogle Scholar
- Han J, Lee J-D, Bibbs L, Ulevitch RJ: A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. Science. 1994, 265: 808-811. 10.1126/science.7914033.View ArticlePubMedGoogle Scholar
- Han J, Sun P: The pathways to tumor suppression via route p38. Trends Biochem Sci. 2007, 32: 364-371. 10.1016/j.tibs.2007.06.007.View ArticlePubMedGoogle Scholar
- Wang XZ, Ron D: Stress-induced phosphorylation and activation of the transcription factor CHOP (GADD153) by p38 MAP kinase. Science. 1996, 272: 1347-1349. 10.1126/science.272.5266.1347.View ArticlePubMedGoogle Scholar
- Kim JY, Choi JA, Kim TH, Yoo YD, Kim JI, Lee YJ, Yoo SY, Cho CK, Lee YS, Lee SJ: Involvement of p38 mitogen-activated protein kinase in the cell growth inhibition by sodium arsenite. J Cell Physiol. 2002, 190: 29-37. 10.1002/jcp.10049.View ArticlePubMedGoogle Scholar
- Chen G, Hitomi M, Han J, Stacey DW: The p38 pathway provides negative feedback for Ras proliferative signaling. J Biol Chem. 2000, 275: 38973-38980. 10.1074/jbc.M002856200.View ArticlePubMedGoogle Scholar
- Molnar A, Theodoras A, Zon L, Kyriakis J: Cdc42Hs, but not rac1, inhibits serum- stimulated cell cycle progression at G1/S through a mechanism requiring p38/RK. J Biol Chem. 1997, 272: 13229-13235. 10.1074/jbc.272.20.13229.View ArticlePubMedGoogle Scholar
- Puri PL, Wu Z, Zhang P, Wood LD, Bhakta KS, Han J, Feramisco JR, Karin M, Wang JYJ: Induction of terminal differentiation by constitutive activation of p38 MAP kinase in human rhabdomyosarcoma cells. Genes Dev. 2011, 14: 574-584.Google Scholar
- Faust D, Dolado I, Cuadrado A, Oesch F, Weiss C, Nebreda AR, Dietrich C: p38alpha MAPK is required for contact inhibition. Oncogene. 2005, 24: 7941-7945. 10.1038/sj.onc.1208948.View ArticlePubMedGoogle Scholar
- Maher P: p38 mitogen-activated protein kinase activation is required for fibroblast growth factor-2-stimulated cell proliferation but not differentiation. J Biol Chem. 1999, 274: 17491-17498. 10.1074/jbc.274.25.17491.View ArticlePubMedGoogle Scholar
- Dixon M, Agius L, Yeaman SJ, Day CP: Inhibition of rat hepatocyte proliferation by transforming growth factor beta and glucagon is associated with inhibition of ERK2 and p70 S6 kinase. Hepatology. 1999, 29: 1418-1424. 10.1002/hep.510290516.View ArticlePubMedGoogle Scholar
- Rausch O, Marshall C: Cooperation of p38 and extracellular signal-regulated kinase mitogen-activated protein kinase pathways during granulocyte colony-stimulating factor-induced hemopoietic cell proliferation. J Biol Chem. 1999, 274: 4096-4105. 10.1074/jbc.274.7.4096.View ArticlePubMedGoogle Scholar
- Chen L, Mayer JA, Krisko TI, Speers CW, Wang T, Hilsenbeck SG, Brown PH: Inhibition of the p38 kinase suppresses the proliferation of human ER-negative breast cancer cells. Cancer Res. 2009, 69: 8853-8861. 10.1158/0008-5472.CAN-09-1636.PubMed CentralView ArticlePubMedGoogle Scholar
- Halawani D, Mondeh R, Stanton LA, Beier F: p38 MAP kinase signaling is necessary for rat chondrosarcoma cell proliferation. Oncogene. 2004, 23: 3726-3731. 10.1038/sj.onc.1207422.View ArticlePubMedGoogle Scholar
- Recio JA, Merlino G: Hepatocyte growth factor/scatter factor activates proliferation in melanoma cells through p38 MAPK, ATF-2 and cyclin D1. Oncogene. 2002, 21: 1000-1008. 10.1038/sj.onc.1205150.View ArticlePubMedGoogle Scholar
- Cuenda A, Rouse J, Doza Y, Meier R, Cohen P, Gallagher T, Young P, Lee J: SB203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett. 1995, 364: 229-233. 10.1016/0014-5793(95)00357-F.View ArticlePubMedGoogle Scholar
- Behren A, Binder K, Vucelic G, Herberhold S, Hirt B, Loewenheim H, Preyer S, Zenner HP, Simon C: The p38 SAPK pathway is required for Ha-ras induced in vitro invasion of NIH3T3 cells. Exp Cell Res. 2005, 303: 321-330. 10.1016/j.yexcr.2004.10.004.View ArticlePubMedGoogle Scholar
- Bain J, Plater L, Elliot M, Shpiro N, Hastie J, McLaughlan H, Klevernic I, Arthur JSC, Alessi DR, Cohen P: The selectivity of protein kinase inhibitors: a further update. Biochem J. 2007, 408: 297-315. 10.1042/BJ20070797.PubMed CentralView ArticlePubMedGoogle Scholar
- Raingeaud J, Whitmarsh A, Barrett T, Derijard B, Davis R: MKK3- and MKK6- regulated gene expression is mediated by the p38 mitogen-activated protein kinase signal transduction pathway. Mol Cell Biol. 1996, 16: 1247-1255.PubMed CentralView ArticlePubMedGoogle Scholar
- Ono K, Han J: The p38 signal transduction pathway Activation and function. Cell Signal. 2000, 12: 1-13. 10.1016/S0898-6568(99)00071-6.View ArticlePubMedGoogle Scholar
- Malumbres M, Barbacid M: Mammalian cyclin-dependent kinases. Trends Biochem Sci. 2005, 30: 630-641. 10.1016/j.tibs.2005.09.005.View ArticlePubMedGoogle Scholar
- Raingeaud J, Gupta S, Rogers J, Dickens M, Han J, Ulevitch RJ, Davis R: Pro- inflammatory cytokines and environmental stress cause p38 mitogen-activated protein kinase activation by dual phosphorylation on tyrosine and threonine. J Biol Chem. 1995, 270: 7420-7426. 10.1074/jbc.270.13.7420.View ArticlePubMedGoogle Scholar
- Deak M, Clifton A, Lucocq J, Alessi D: Mitogen- and stress-activated protein kinase-1 (MSK1) is directly activated by MAPK and SAPK2/p38, and may mediate activation of CREB. EMBO J. 1998, 17: 4426-4441. 10.1093/emboj/17.15.4426.PubMed CentralView ArticlePubMedGoogle Scholar
- Tan Y, Rouse J, Zhang A, Cariati S, Cohen P, Comb M: FGF and stress regulate CREB and ATF-1 via a pathway involving p38 MAP kinase and MAPKAP-K2. EMBO J. 1996, 15: 4629-4642.PubMed CentralPubMedGoogle Scholar
- Iordanov M, Bender K, Ade T, Schmid W, Sachsenmaier C, Engel K, Gaestel M, Rahmsdorf HJ, Herrlich P: CREB is activated by UVC through a p38/HOG-1- dependent protein kinase. EMBO J. 1997, 16: 1009-1022. 10.1093/emboj/16.5.1009.PubMed CentralView ArticlePubMedGoogle Scholar
- Aktories K: Rho proteins: targets for bacterial toxins. TiMB. 1997, 5: 282-288.Google Scholar
- Aktories K, Barbieri JT: Bacterial cytotoxins: targeting eukaryotic switches. Nat Rev Microbiol. 2005, 3: 397-410. 10.1038/nrmicro1150.View ArticlePubMedGoogle Scholar
- Barbieri JT, Riese MJ, Aktories K: Bacterial toxins that modify the actin cytoskeleton. Annu Rev Cell Dev Biol. 2002, 18: 315-344. 10.1146/annurev.cellbio.18.012502.134748.View ArticlePubMedGoogle Scholar
- Popoff MR, Chaves-Olarte E, Lemichez E, von Eichel-Streiber C, Thelestam M, Chardin P, Cussac D, Antonny B, Chavrier P, Flatau G, Giry M, de Gunzburg J, Boquet P: Ras, Rap, and Rac small GTP-binding proteins are targets for Clostridium sordellii lethal toxin glucosylation. J Biol Chem. 1996, 271: 10217-10224. 10.1074/jbc.271.17.10217.View ArticlePubMedGoogle Scholar
- Marshall C: Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell. 1995, 80: 179-185. 10.1016/0092-8674(95)90401-8.View ArticlePubMedGoogle Scholar
- Lee RJ, Albanese C, Stenger RJ, Watanabe G, Inghirami G, Haines GK, Webster M, Muller WJ, Brugge JS, Davis RJ, Pestell RG: pp 60(v-src) induction of cyclin D1 requires collaborative interactions between the extracellular signal-regulated kinase, p38, and Jun kinase pathways. A role for cAMP response element-binding protein and activating transcription factor-2 in pp60(v-src) signaling in breast cancer cells. J Biol Chem. 1999, 274: 7341-7350. 10.1074/jbc.274.11.7341.View ArticlePubMedGoogle Scholar
- Villanueva J, Yung Y, Walker JL, Assoian RK: ERK activity and G1 phase progression: identifying dispensable versus essential activities and primary versus secondary targets. Mol Biol Cell. 2007, 18: 1457-1463. 10.1091/mbc.E06-10-0908.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang W, Chen JX, Liao R, Deng Q, Zhou J, Huang S, Sun P: Sequential activation of the MEK-extracellular signal-related kinase and MKK3/6-p38 mitogen-activated protein kinase pathways mediates oncogenic ras-induced premature senescence. Mol Cell Biol. 2002, 22: 2289-3403.Google Scholar
- Iwasa H, Han J, Ishikawa F: Mitogen-activated protein kinase p38 defines the common senescence-signalling pathway. Genes Cells. 2003, 8: 131-144. 10.1046/j.1365-2443.2003.00620.x.View ArticlePubMedGoogle Scholar
- Behren A, Mühlen S, Acuna Sanhueza GA, Schwager C, Plinkert PK, Huber PE, Abdollahi A, Simon C: Phenotype-assisted transcriptome analysis identifies foxm1 downstream from ras-MKK3-p38 to regulate in vitro cellular invasion. Oncogene. 2010, 29: 1519-1530. 10.1038/onc.2009.436.View ArticlePubMedGoogle Scholar
- Halasi M, Schraufnagel DP, Gartel AL: Wild-type p53 protects normal cells against apoptosis induced by thiostrepton. Cell Cycle. 2009, 8: 2850-2851. 10.4161/cc.8.17.9414.View ArticlePubMedGoogle Scholar
- Bhana S, Hewer A, Phillips DH, Lloyd DR: p53-dependent global nucleotide excision repair of cisplatin-induced intrastrand cross links in human cells. Mutagenesis. 2008, 23: 131-136. 10.1093/mutage/gen001.View ArticlePubMedGoogle Scholar
- Hazzalin C, Cano E, Cuenda A, Barratt M, Cohen P, Mahadevan L: p38/RK is essential for stress-induced nuclear responses: JNK/SAPKs and c-jun/ATF-2 phosphorylation are insufficient. Curr Biol. 1996, 6: 1028-1031. 10.1016/S0960-9822(02)00649-8.View ArticlePubMedGoogle Scholar
- Lafarga V, Cuadrado A, Lopez de Silanes I, Bengoechea R, Fernandez-Capetillo O, Nebreda AR: p38 Mitogen-activated protein kinase- and HuR-dependent stabilization of p21(Cip1) mRNA mediates the G(1)/S checkpoint. Mol Cell Biol. 2009, 29: 4341-4351. 10.1128/MCB.00210-09.PubMed CentralView ArticlePubMedGoogle Scholar
- Cuadrado M, Gutierrez-Martinez P, Swat A, Nebreda AR, Fernandez-Capetillo O: p27Kip1 stabilization is essential for the maintenance of cell cycle arrest in response to DNA damage. Cancer Res. 2009, 69: 8726-8732. 10.1158/0008-5472.CAN-09-0729.PubMed CentralView ArticlePubMedGoogle Scholar
- Swat A, Dolado I, Rojas JM, Nebreda AR: Cell density-dependent inhibition of epidermal growth factor receptor signaling by p38alpha mitogen-activated protein kinase via Sprouty2 downregulation. Mol Cell Biol. 2009, 29: 3332-3343. 10.1128/MCB.01955-08.PubMed CentralView ArticlePubMedGoogle Scholar
- Luo Y, Zou P, Zou J, Wang J, Zhou D, Liu L: Autophagy regulates ROS-induced cellular senescence via p21 in a p38 MAPKα dependent manner. Exp Gerontol. 2011, 46: 860-867. 10.1016/j.exger.2011.07.005.PubMed CentralView ArticlePubMedGoogle Scholar
- Rudolf E, Cervinka M: Stress responses of human dermal fibroblasts exposed to zinc pyrithione. Toxicol Lett. 2011, 204: 164-173. 10.1016/j.toxlet.2011.04.028.View ArticlePubMedGoogle Scholar
- Nakagawa H, Hirata Y, Takeda K, Hayakawa Y, Sato T, Kinoshita H, Sakamoto K, Nakata W, Hikiba Y, Omata M, Yoshida H, Koike K, Ichijo H, Maeda S: Apoptosis signal-regulating kinase 1 inhibits hepatocarcinogenesis by controlling the tumor- suppressing function of stress-activated mitogen-activated protein kinase. Hepatology. 2011, 54: 185-195. 10.1002/hep.24357.View ArticlePubMedGoogle Scholar
- Shao L, Li H, Pazhanisamy SK, Meng A, Wang Y, Zhou D: Reactive oxygen species and hematopoietic stem cell senescence. Int J Hematol. 2011, 94: 24-32. 10.1007/s12185-011-0872-1.PubMed CentralView ArticlePubMedGoogle Scholar
- Li H, Wang W, Liu X, Paulson KE, Yee AS, Zhang X: Transcriptional factor HBP1 targets P16(INK4A), upregulating its expression and consequently is involved in Ras- induced premature senescence. Oncogene. 2010, 29: 5083-5094. 10.1038/onc.2010.252.View ArticlePubMedGoogle Scholar
- Kwong J, Hong L, Liao R, Deng Q, Han J, Sun P: p38alpha and p38gamma mediate oncogenic ras-induced senescence through differential mechanisms. J Biol Chem. 2009, 284: 11237-11246.PubMed CentralView ArticlePubMedGoogle Scholar
- Bulavin DV, Phillips C, Nannenga B, Timofeev O, Donehower LA, Anderson CW, Appella E, Fornace AJ: Inactivation of the Wip1 phosphatase inhibits mammary tumorigenesis through p38 MAPK-mediated activation of the p16(Ink4a)-p19(Arf) pathway. Nat Genet. 2004, 36: 343-350. 10.1038/ng1317.View ArticlePubMedGoogle Scholar
- Mavrogonatou E, Kletsas D: Differential response of nucleus pulposus intervertebral disc cells to high salt, sorbitol and urea. J Cell Physiol. 2011, doi:10.1002/jcp.22840Google Scholar
- Farini D, Puglianiello A, Mammi C, Siracusa G, Moretti C: Dual effect of pituitary adenylate cyclase activating polypeptide on prostate tumor LNCaP cells: short- and long-term exposure affect proliferation and neuroendocrine differentiation. Endocrinology. 2003, 144: 1631-1643. 10.1210/en.2002-221009.View ArticlePubMedGoogle Scholar
- Gray MC, Hewlett EL: Cell cycle arrest induced by the bacterial adenylate cyclase toxins from Bacillus anthracis and Bordetella pertussis. Cell Microbiol. 2011, 131: 123-134.View ArticleGoogle Scholar
- Li Y, Yin W, Wang X, Zhu W, Huang Y, Yan G: Cholera toxin induces malignant glioma cell differentiation via the PKA/CREB pathway. Proc Natl Acad Sci USA. 2007, 104: 13438-13443. 10.1073/pnas.0701990104.PubMed CentralView ArticlePubMedGoogle Scholar
- Shen G, Xu C, Chen C, Hebbar V, Kong AN: p53-independent G1 cell cycle arrest of human colon carcinoma cells HT-29 by sulforaphane is associated with induction of p21CIP1 and inhibition of expression of cyclin D1. Cancer Chemother Pharmacol. 2006, 57: 317-327. 10.1007/s00280-005-0050-3.View ArticlePubMedGoogle Scholar
- van Dam H, Wilhelm D, Herr I, Steffen A, Herrlich P, Angel P: ATF-2 is preferentially activated by stress-activated protein kinases to mediate c-jun induction in response to genotoxic agents. EMBO J. 1995, 14: 1798-1811.PubMed CentralPubMedGoogle Scholar
- Cavigelli M, Li W, Lin A, Su B, Yoshioka K, Karin M: The tumor promoter arsenite stimulates AP-1 activity by inhibiting a JNK phosphatase. EMBO J. 1996, 15: 6269-6279.PubMed CentralPubMedGoogle Scholar
- Coso O, Teramoto H, Simonds W, Gutkind JS: Signaling from G protein-coupled receptors to c-jun kinase involves bg subunits or heterotrimeric G proteins acting on a ras and rac1-dependent pathway. J Biol Chem. 1995, 271: 3963-3966.Google Scholar
- Barth H, Olenik C, Sehr P, Schmidt G, Aktories K, Meyer DK: Neosynthesis and activation of rho by Escherichia coli cytotoxic necrotizing factor (CNF1) reverse cytopathic effects of ADP-ribosylated rho. J Biol Chem. 1999, 274: 27407-27414. 10.1074/jbc.274.39.27407.View ArticlePubMedGoogle Scholar
- Bagrodia S, Derijard B, Davis R, Cerione R: Cdc42 and PAK-mediated signaling leads to jun kinase and p38 mitogen-activated protein kinase activation. J Biol Chem. 1995, 270: 27995-27998. 10.1074/jbc.270.47.27995.View ArticlePubMedGoogle Scholar
- Minden A, Lin A, Claret F-X, Abo A, Karin M: Selective activation of the JNK signaling cascade and c-jun transcriptional activity by the small GTPases rac and cdc42Hs. Cell. 1995, 81: 1147-1157. 10.1016/S0092-8674(05)80019-4.View ArticlePubMedGoogle Scholar
- Dietrich C, Wallenfang K, Oesch F, Wieser R: Differences in the mechanisms of growth control in contact-inhibited and serum-deprived human fibroblasts. Oncogene. 1997, 15: 2743-2747. 10.1038/sj.onc.1201439.View ArticlePubMedGoogle Scholar
- Dietrich C, Bartsch T, Schanz F, Oesch F, Wieser R: p53-dependent cell cycle arrest induced by N-acetyl-L-leucinyl-L-norleucinal in platelet-derived growth factor stimulated human fibroblasts. Proc Natl Acad Sci USA. 1996, 93: 10815-10819. 10.1073/pnas.93.20.10815.PubMed CentralView ArticlePubMedGoogle Scholar
- Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC: Measurement of protein using bicinchoninic acid. Anal Biochem. 1985, 150: 76-78. 10.1016/0003-2697(85)90442-7.View ArticlePubMedGoogle Scholar
- Weiss C, Faust D, Dürk H, Kolluri SK, Pelzer A, Schneider S, Dietrich C, Oesch F, Göttlicher M: TCDD induces c-jun expression via a novel Ah (dioxin) receptor-mediated p38-MAPK-dependent pathway. Oncogene. 2005, 24: 4975-4983. 10.1038/sj.onc.1208679.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.