Examining the mechanism of Erk nuclear translocation using green fluorescent protein

Horgan, Angela; Stork, Philip J.S.

doi:10.1016/s0014-4827(03)00037-5

Cited by 58 publications

(46 citation statements)

References 48 publications

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“…The Dynamics of Localization of ERK1/2 Not Complexed with MEK1-Similar to what has been described for cell lines (27,46), we found that in unstimulated hippocampal neurons coexpression of MEK1 is required to restrict the localization of ERK1/2-Dronpa and ERK2-GFP to the cytoplasm when the ERK fusion proteins are overexpressed in large quantities. In the absence of exogenously expressed MEK1, ERK1/2-Dronpa and ERK2-GFP localized to both the cytoplasm and the nucleus, a distribution pattern similar to the passively diffusing proteins Dronpa and MBP-Dronpa.…”

Section: Discussionsupporting

confidence: 83%

Diffusion and Not Active Transport Underlies and Limits ERK1/2 Synapse-to-Nucleus Signaling in Hippocampal Neurons

Wiegert

Bengtson

Bading

2007

Journal of Biological Chemistry

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The propagation of signals from synapses and dendrites to the nucleus is crucial for long lasting adaptive changes in the nervous system. The ERK-MAPK pathway can link neuronal activity and cell surface receptor activation to the regulation of gene transcription, and it is often considered the principal mediator of synapse-to-nucleus communication in late-phase plasticity and learning. However, the mechanisms underlying ERK1/2 trafficking in dendrites and nuclear translocation in neurons remain to be determined leaving it unclear whether ERK1/2 activated at the synapse can contribute to nuclear signaling and transcriptional regulation. Using the photobleachable and photoactivable fluorescent tag Dronpa on ERK1 and ERK2, we show here that ERK1/2 translocation to the nucleus of hippocampal neurons is induced by the stimulation of N-methyl-D-aspartate receptors or TrkB stimulation and is apparently mediated by facilitated diffusion. In contrast, ERK1/2 trafficking within dendrites is not signal-regulated and is mediated by passive diffusion. Within dendrites, the reach of a locally activated pool of ERK1/2 is very limited and follows an exponential decay with distance. These results indicate that successful signal propagation to the nucleus by the ERK-MAPK pathway depends on the distance of the nucleus from the site of ERK1/2 activation. ERK1/2 activated within or near the soma may rapidly reach the nucleus to induce gene expression, whereas ERK1/2 activated at distal synapses may only contribute to local signaling.The extracellular signal-regulated kinase/mitogen-activated protein (ERK-MAP) 2 kinase cascade is a multifunctional signaling module that plays an important role in the development, differentiation, and proliferation of many cell types. In the nervous system, ERK1 and ERK2 are activated by trophic factors, such as BDNF (1) and electrical activity causing calcium influx into neurons through NMDA receptors or voltage-gated calcium channels (2, 3). A large body of literature has implicated the ERK-MAP kinase pathway in numerous activity-driven neuronal processes, including neuronal survival, synaptic plasticity and learning (4, 5), circadian rhythms (6), pain (7), and addiction (8). ERK1/2 can act locally near the site of activation by cell surface receptors or ion channels to control, for example, the trafficking of ␣-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (9). Another putative role of ERK1/2 in the nervous system is to propagate signals generated at the synapse along the dendrites toward and into the cell nucleus, where it leads to induction of gene transcription. ERK1/2 can phosphorylate several transcription factors, including the ternary complex factors Elk-1 (10, 11) and SAP-1/2 (12), as well as c-Jun (13) and c-Fos (14). The cAMP-response element-binding protein (CREB) is also a target of the ERK-MAP kinase pathway, although CREB phosphorylation on its activator site serine 133 is catalyzed not by ERK1/2 but by the ERK1/2-regulated kinases ribosomal protein S6 kinase 2 and mitog...

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Section: Discussionsupporting

confidence: 83%

Diffusion and Not Active Transport Underlies and Limits ERK1/2 Synapse-to-Nucleus Signaling in Hippocampal Neurons

Wiegert

Bengtson

Bading

2007

Journal of Biological Chemistry

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“…On the contrary, unpaired conditioning requires recruitment of sustained neuronal activity in the hippocampus that is accompanied by a prolonged ERK1/2 phosphorylation possibly triggered by the PKA signaling pathway. In both conditioning situations, the first wave of phosphorylation of ERK1/2 is sufficient to stimulate various cellular targets, such as CREB and Elk-1, leading to transcriptional modulation of immediate early genes that can initiate long-lasting forms of synaptic plasticity (Sgambato et al 1998;Vanhoutte et al 1999;Horgan and Stork 2003). Our results provide further evidence that inhibition of the first wave of ERK1/2 activation dramatically impaired long-term retention without affecting encoding and short-term memory processes.…”

Section: Learning and Memory 353supporting

confidence: 54%

Foreground contextual fear memory consolidation requires two independent phases of hippocampal ERK/CREB activation

Trifilieff¹,

Herry²,

Vanhoutte³

et al. 2006

Learn. Mem.

138

124

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Fear conditioning is a popular model for investigating physiological and cellular mechanisms of memory formation. In this paradigm, a footshock is either systematically associated to a tone (paired conditioning) or is pseudorandomly distributed (unpaired conditioning). In the former procedure, the tone/shock association is acquired, whereas in the latter procedure, the context/shock association will prevail. Animals with chronically implanted recording electrodes show enhanced amplitude of the extracellularly recorded field EPSP in CA1 pyramidal cells for up to 24 h after unpaired, but not paired, fear conditioning. This is paralleled by a differential activation of the ERK/CREB pathway in CA1, which is monophasic in paired conditioning (0-15 min post-conditioning), but biphasic (0-1 h and 9-12 h post-conditioning) in unpaired conditioning as revealed by immunocytochemistry and Western blotting. Intrahippocampal injection of the MEK inhibitor U0126 prior to each phase prevents the activation of both ERK1/2 and CREB after unpaired conditioning. Block of any activation phase leads to memory impairment. We finally reveal that the biphasic activation of ERK/CREB activity is independently regulated, yet both phases are critically required for the consolidation of long-term memories following unpaired fear conditioning. These data provide compelling evidence that CA1 serves different forms of memory by expressing differential cellular mechanisms that are dependent on the training regime.

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“…Full-length cDNA of RPS6KA genes were released from the pMT2 vector using EcoRI digestion and subcloned with or without a HA tag (as a linker with overhangs for restriction sites KpnI and BamHI, (CATG-TACCCATAC-GATGTTCCAGATTACGCTGGG) into the eukaryotic expression vectors, pcDNA3/neo, pcDNA3.1, pcDNA4/TO and pcDNA5/TO (Invitrogen, Paisley, Strathclyde, UK), and pEGFP-C3 (BD Biosciences). GFP-Erk2a was a gift of Dr Philip Stork (Horgan and Stork, 2003).…”

Section: Methodsmentioning

confidence: 99%

RPS6KA2, a putative tumour suppressor gene at 6q27 in sporadic epithelial ovarian cancer

et al. 2006

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We had previously defined by allele loss studies a minimal region at 6q27 (between D6S264 and D6S297) to contain a putative tumour suppressor gene. The p90 ribosomal S6 kinase-3 gene (p90 Rsk-3, RPS6KA2) maps in this interval. It is a serine-threonine kinase that signals downstream of the mitogen-activated protein kinase pathway. It is expressed in normal ovarian epithelium, whereas reduced or absent in tumours or cell lines. We show that RPS6KA2 is monoallelically expressed in the ovary suggesting that loss of a single expressed allele is sufficient to cause complete loss of expression in cancer cells. Further, we have identified two new isoforms of RPS6KA2 with an alternative start codon. Homozygous deletions were identified within the RPS6KA2 gene in two cell lines. Re-expression of RPS6KA2 in ovarian cancer cell lines suppressed colony formation. In UCI101 cells, the expression of RPS6KA2 reduced proliferation, caused G1 arrest, increased apoptosis, reduced levels of phosphorylated extracellular signal-regulated kinase and altered other cell cycle proteins. In contrast, small interfering RNA against RPS6KA2 showed the opposite effect in 41M cells. The above results suggest that RPS6KA2 is a putative tumour suppressor gene to explain allele loss at 6q27.

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Examining the mechanism of Erk nuclear translocation using green fluorescent protein

Cited by 58 publications

References 48 publications

Diffusion and Not Active Transport Underlies and Limits ERK1/2 Synapse-to-Nucleus Signaling in Hippocampal Neurons

Diffusion and Not Active Transport Underlies and Limits ERK1/2 Synapse-to-Nucleus Signaling in Hippocampal Neurons

Foreground contextual fear memory consolidation requires two independent phases of hippocampal ERK/CREB activation

RPS6KA2, a putative tumour suppressor gene at 6q27 in sporadic epithelial ovarian cancer

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