MLCK‐independent phosphorylation of MLC20 and its regulation by MAP kinase pathway in human bladder smooth muscle cells

Deng, Maoxian; Ding, Wei; Min, Xuewen; Xia, Ying

doi:10.1002/cm.20471

Cited by 20 publications

(26 citation statements)

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“…For example, inhibition of ERK by inhibitor and antisense oligonucleotide blocks smooth muscle migration (Graf et al, 1997;Gerthoffer, 2007). Our study confirmed role of ERK1/2 in smooth muscle cell migration by blocking movement by PD-098059 (Deng et al, 2011). In addition, Raf-1 kinase, the upstream molecule of MEK1/2 that activates ERK1/2, may be involved in regulation of smooth muscle contraction (Sathishkumar et al, 2010).…”

Section: Erk Pathwaysupporting

confidence: 76%

“…In the studies, the hBSM cells were exposed to different concentrations of the MAP kinase inhibitors for 40 minutes, followed by Western blotting assays for the phosphor MLC20. The results show that phosphor MLC20 level increases with the doses of inhibitors ( Figure 6C and D, ERK inhibitor not shown) (Deng et al, 2011). A similar method was used to examine the effects of the MEKK1 at different infection doses (i.e., pfu/cell) on the MLC20 phosphorylation.…”

Section: Activation Of Map Kinase Pathways Inhibits Mlc20 Phosphorylamentioning

confidence: 90%

“…Cells for ERK inhibition were previously permeabilized (Deng et al, 2011). Proteins of cell lysates were separated by SDS-PAGE electrophoresis and then transferred to PVDF membranes.…”

Section: Inhibition Of Map Kinase Pathways Increases Phosphorylation mentioning

confidence: 99%

“…Then, the MEKK1 adenovirus was used for the evaluation of the roles of the MAP kinase on MLC20 phosphorylation. When the cell density was between 60-70%, the hBSM cells in 6-well plates were infected with the MEKK1 and the control viruses at different doses (Deng et al, 2011). Three days after infection, the cells were harvested for Western blotting.…”

Section: Activation Of Map Kinase Pathways Inhibits Mlc20 Phosphorylamentioning

confidence: 99%

“…The viral expressive efficiency of MEKK1 was assayed in an independent experiment by Western blotting using an antibody against the HA-Tag (Deng et al, 2011). The exogenous MEKK1 activity expressed by the adenovirus was estimated by its capacity to activate the downstream molecules, including the JNK, ERK, and p38.…”

Section: Activation Of Map Kinase Pathways Inhibits Mlc20 Phosphorylamentioning

confidence: 99%

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MAP Kinase-Mediated and MLCK-Independent Phosphorylation of MLC20 in Smooth Muscle Cells

Deng¹,

Deng²,

Xue³

2012

Current Basic and Pathological Approaches to the Function of Muscle Cells and Tissues - From Molecules to Humans

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Section: Erk Pathwaysupporting

confidence: 76%

Section: Activation Of Map Kinase Pathways Inhibits Mlc20 Phosphorylamentioning

confidence: 90%

“…Cells for ERK inhibition were previously permeabilized (Deng et al, 2011). Proteins of cell lysates were separated by SDS-PAGE electrophoresis and then transferred to PVDF membranes.…”

Section: Inhibition Of Map Kinase Pathways Increases Phosphorylation mentioning

confidence: 99%

Section: Activation Of Map Kinase Pathways Inhibits Mlc20 Phosphorylamentioning

confidence: 99%

Section: Activation Of Map Kinase Pathways Inhibits Mlc20 Phosphorylamentioning

confidence: 99%

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MAP Kinase-Mediated and MLCK-Independent Phosphorylation of MLC20 in Smooth Muscle Cells

Deng¹,

Deng²,

Xue³

2012

Current Basic and Pathological Approaches to the Function of Muscle Cells and Tissues - From Molecules to Humans

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TCR signalling network organization at the immunological synapses of murine regulatory T cells

et al. 2017

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Regulatory T (Treg) cells require T-cell receptor (TCR) signalling to exert their immunosuppressive activity, but the precise organization of the TCR signalling network compared to conventional T (Tconv) cells remains elusive. By using accurate mass spectrometry and multi-epitope ligand cartography (MELC) we characterized TCR signalling and recruitment of TCR signalling components to the immunological synapse (IS) in www.eji-journal.euThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. 2Marco van Ham et al. Eur. J. Immunol. 2017. 0: 1-16 in Treg cells, thereby, seems to be differentially organized as in Tconv cells: Treg cells show reduced Ca 2+ flux and ERK1/2 phosphorylation upon TCR stimulation [6][7][8], and downstream signalling molecules such as Lck, LAT and PLCγ1 are essential for Treg cell suppressive capacity [9,10]. In this line, a novel TCRmediated ADAP/integrin-independent PLCγ1 activation pathway was described to be required for suppression of Tconv cells by Treg cells [11]. Furthermore, Treg cells exhibit reduced S473 phosphorylation of Akt which seems a prerequisite for suppression [12]. Although the enzymatic activity of the tyrosine kinase ZAP70 seems to be dispensable for the suppressive phenotype of Foxp3 + Treg cells [13], a single mutation within the SH2-domain of ZAP70 leads to impaired suppressive activities, which indicated its importance at the immunological synapse (IS) [14]. Finally, it has only recently been recognized that the recruitment of TCR signalling components into the IS is differentially organized in Treg cells and Tconv cells. The protein kinase PKCθ is recruited to the IS in Tconv cells, in Treg cells, however, this kinase was found to be sequestered away from the IS, but still of importance to control suppression [15]. The spatial recruitment of signalling components during IS formation critically depends on cytoskeleton dynamics, which in turn is controlled by TCR activation and subsequent phosphorylation of proteins regulating cytoskeleton reorganization [5]. As part of these processes the microtubule-organizing centre (MTOC) is rapidly translocated in proximity to the IS. MTOC repositioning depends on LAT, ZAP70 and SLP76 [16] and is regulated by a cascade of distinct isoforms of the family of novel protein kinase C (nPKC) [17]. The MTOC serves as a platform to coordinate molecular movements from and to the IS through the support of microtubule (MT) motors [18]. For instance, TCR microclusters move along MTs towards the centre of the IS in a dynein-dependent manner [19], and hindrance of MTOC polarization, molecular transport and cytoskeleton dynamics prevent proper propagation of TCR signals [20].It is now tempting to speculate that the localization of signalling modules within the IS may be instrumental for the formation of a Treg cell-specific IS and the suppressive phenotype of Treg cells. At this moment, however,...

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Signaling pathways in cytoskeletal responses to plasma membrane depolarization in corneal endothelial cells

Evans

Hernández

Chifflet

2019

Journal Cellular Physiology

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In previous work, we reported that plasma membrane potential depolarization (PMPD) provokes cortical F-actin remodeling in bovine corneal endothelial (BCE) cells in culture, which eventually leads to the appearance of intercellular gaps. In kidney epithelial cells it has been shown that PMPD determines an extracellularsignal-regulated kinase (ERK)/Rho-dependent increase in diphosphorylated myosin light chain (ppMLC). The present study investigated the signaling pathways involved in the response of BCE cells to PMPD. Differently to renal epithelial cells, we observed that PMPD leads to a decrease in monophosphorylated MLC (pMLC) without affecting diphosphorylated MLC. Also, that the pMLC reduction is a consequence of cyclic adenosine 3′,5′-monophosphate (cAMP)/protein kinase A (PKA) activation. In addition, we found evidence that the cAMP increase mostly depends on soluble adenylyl cyclase activity. Inhibition of this enzyme reduces the effect of PMPD on the cAMP rise, F-actin remodeling, and pMLC decrease. No changes in phosho-ERK were observed, although we could determine that RhoA undergoes activation. Our results suggested that active RhoA is not involved in the intercellular gap formation. Overall, the findings of this study support the view that, differently to renal epithelial cells, in BCE cells PMPD determines cytoskeletal reorganization via activation of the cAMP/PKA pathway. K E Y W O R D S actin cytoskeleton, cAMP/PKA, corneal endothelium, myosin light chain, plasma membrane depolarization

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MLCK‐independent phosphorylation of MLC20 and its regulation by MAP kinase pathway in human bladder smooth muscle cells

Cited by 20 publications

References 58 publications

MAP Kinase-Mediated and MLCK-Independent Phosphorylation of MLC20 in Smooth Muscle Cells

MAP Kinase-Mediated and MLCK-Independent Phosphorylation of MLC20 in Smooth Muscle Cells

TCR signalling network organization at the immunological synapses of murine regulatory T cells

Signaling pathways in cytoskeletal responses to plasma membrane depolarization in corneal endothelial cells

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