ERK2 but not ERK1 mediates HGF-induced motility in non-small cell lung carcinoma cell lines

TY - JOUR

T1 - ERK2 but not ERK1 mediates HGF-induced motility in non-small cell lung carcinoma cell lines

AU - Radtke, Simone

AU - Milanovic, Mina

AU - Rosse, Carine

AU - De Rycker, Manu

AU - Lachmann, Sylvie

AU - Hibbert, Andrew

AU - Kermorgant, Stephanie

AU - Parker, Peter

PY - 2013/6/1

Y1 - 2013/6/1

N2 - Aberrant signalling of receptor tyrosine kinases (RTKs), such as c-Met, the receptor for hepatocyte growth factor (HGF), has been implicated in the oncogenesis of various tumours including non-small cell lung carcinoma (NSCLC). Through its pro-migratory properties, c-Met has been implicated specifically in the process of tumour metastasis, demanding a better understanding of the underlying signalling pathways. Various players downstream of c-Met have been well characterised, including the extracellular-signal-regulated kinases (ERKs) 1 and 2. In a small interfering RNA (siRNA)-based high-throughput wound healing screen performed in A549 lung carcinoma cells, we identified ERK2 but not ERK1 as a strong mediator of HGF-induced motility. This finding was confirmed in several NSCLC cell lines as well as in HeLa cells. One known substrate for ERK kinases in cell migration, the focal adhesion protein paxillin, was also one of the hits identified in the screen. We demonstrate that HGF stimulation results in a time-dependent phosphorylation of paxillin on serine 126, a process that can be blocked by inhibition of the ERK1/2 upstream kinase mitogen-activated protein kinase/ERK kinase 1 (MEK1) or inhibition of glycogen synthase kinase 3 (GSK3). Further, we show that paxillin turnover at focal adhesions is increased upon stimulation by HGF, an effect that is dependent on serine residues 126 (GSK3 site) and 130 (ERK site) within paxillin. In line with the isoform-specific requirement of ERK2 for HGF-mediated migration in lung tumour cell models, ERK2 but not ERK1 is shown to be responsible for paxillin serine 126 phosphorylation and its increased turnover at focal adhesions.

AB - Aberrant signalling of receptor tyrosine kinases (RTKs), such as c-Met, the receptor for hepatocyte growth factor (HGF), has been implicated in the oncogenesis of various tumours including non-small cell lung carcinoma (NSCLC). Through its pro-migratory properties, c-Met has been implicated specifically in the process of tumour metastasis, demanding a better understanding of the underlying signalling pathways. Various players downstream of c-Met have been well characterised, including the extracellular-signal-regulated kinases (ERKs) 1 and 2. In a small interfering RNA (siRNA)-based high-throughput wound healing screen performed in A549 lung carcinoma cells, we identified ERK2 but not ERK1 as a strong mediator of HGF-induced motility. This finding was confirmed in several NSCLC cell lines as well as in HeLa cells. One known substrate for ERK kinases in cell migration, the focal adhesion protein paxillin, was also one of the hits identified in the screen. We demonstrate that HGF stimulation results in a time-dependent phosphorylation of paxillin on serine 126, a process that can be blocked by inhibition of the ERK1/2 upstream kinase mitogen-activated protein kinase/ERK kinase 1 (MEK1) or inhibition of glycogen synthase kinase 3 (GSK3). Further, we show that paxillin turnover at focal adhesions is increased upon stimulation by HGF, an effect that is dependent on serine residues 126 (GSK3 site) and 130 (ERK site) within paxillin. In line with the isoform-specific requirement of ERK2 for HGF-mediated migration in lung tumour cell models, ERK2 but not ERK1 is shown to be responsible for paxillin serine 126 phosphorylation and its increased turnover at focal adhesions.

KW - HGF

KW - c-Met

KW - Motility

KW - Wound healing

KW - High-throughput screen

KW - Signalling

KW - siRNA

KW - ERK2

KW - Paxillin

KW - C-MET

KW - EPITHELIAL MORPHOGENESIS

KW - PAXILLIN PHOSPHORYLATION

KW - SIGNALING PATHWAY

KW - TYROSINE KINASE

KW - GROWTH

KW - MIGRATION

KW - CANCER

KW - EXPRESSION

KW - ACTIVATION

U2 - 10.1242/jcs.115832

DO - 10.1242/jcs.115832

M3 - Article

SN - 0021-9533

VL - 126

SP - 2381

EP - 2391

JO - Journal of Cell Science

JF - Journal of Cell Science

IS - 11

ER -