ERF Nuclear Shuttling, a Continuous Monitor of Erk Activity That Links It to Cell Cycle Progression

Gallic, Lionel Le; Virgilio, Laura; Cohen, Philip; Biteau, Benoît; Mavrothalassitis, George

doi:10.1128/mcb.24.3.1206-1218.2004

Cited by 70 publications

(98 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This is an important question because a stabilized repressor may increase the chromatin resident time of its associated chromatin modifiers. In some cases, temporal control of stability is achieved by mechanisms that shuttle the repressors between the nucleus and cytoplasm (29)(30)(31). Although REST has been observed in some cellular contexts in the cytoplasm (28,31,32), its half-life in this cell compartment has not been measured, and whether its cytoplasmic complexes contain kinases or phosphatases has not been determined.…”

Section: Discussionmentioning

confidence: 99%

C-terminal domain small phosphatase 1 and MAP kinase reciprocally control REST stability and neuronal differentiation

Nesti

Corson

McCleskey

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The repressor element 1 (RE1) silencing transcription factor (REST) in stem cells represses hundreds of genes essential to neuronal function. During neurogenesis, REST is degraded in neural progenitors to promote subsequent elaboration of a mature neuronal phenotype. Prior studies indicate that part of the degradation mechanism involves phosphorylation of two sites in the C terminus of REST that require activity of beta-transducin repeat containing E3 ubiquitin protein ligase, βTrCP. We identify a proline-directed phosphorylation motif, at serines 861/864 upstream of these sites, which is a substrate for the peptidylprolyl cis/trans isomerase, Pin1, as well as the ERK1/2 kinases. Mutation at S861/864 stabilizes REST, as does inhibition of Pin1 activity. Interestingly, we find that C-terminal domain small phosphatase 1 (CTDSP1), which is recruited by REST to neuronal genes, is present in REST immunocomplexes, dephosphorylates S861/864, and stabilizes REST. Expression of a REST peptide containing S861/864 in neural progenitors inhibits terminal neuronal differentiation. Together with previous work indicating that both REST and CTDSP1 are expressed to high levels in stem cells and down-regulated during neurogenesis, our results suggest that CTDSP1 activity stabilizes REST in stem cells and that ERK-dependent phosphorylation combined with Pin1 activity promotes REST degradation in neural progenitors.T he repressor element 1 (RE1) silencing transcription factor (REST) is a transcriptional repressor that suppresses neuronal gene expression in nonneural cells, such as fibroblasts, as well as in neural progenitors (1-3). Its targets represent genes required for the terminally differentiated neuronal cell phenotype, including genes encoding voltage and ligand-dependent ion channels, receptors, growth factors, and axonal-guidance proteins (4-7). Thus, during neurogenesis, REST is progressively down-regulated to allow elaboration of the mature neuronal phenotype (3). Nonetheless, precisely how REST itself is regulated still remains an open question. Relatively little is known about either its transcriptional or posttranscriptional regulation (3,8,9). In contrast, several studies have focused on posttranslational regulation of REST (3, 10, 11), but the identity of the signaling molecules involved has received little attention.In neural progenitors and human embryonic kidney (HEK) cells, rapid REST turnover is mediated by targeting to a proteasomal pathway (3, 10, 11). REST degradation during neuronal differentiation in culture requires interaction with beta-transducin repeat containing E3 ubiquitin protein ligase (βTrCP) for targeting to the proteasome (11). βTrCP was also required for cell-cycle-dependent degradation of REST in HEK cells (10). Two adjacent phosphorylated peptides in the C-terminal domain of REST were identified as βTrCP substrates in these studies, and function as degrons. One kinase responsible for the phosphorylation and degron activity in hippocampus is casein kinase 1, CK1 (12), but whether it function...

show abstract

Section: Discussionmentioning

confidence: 99%

C-terminal domain small phosphatase 1 and MAP kinase reciprocally control REST stability and neuronal differentiation

Nesti

Corson

McCleskey

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Having demonstrated the ability of ERF to associate with the N-box/EBS site of the utrophin-A promoter using gel shift assays, we asked whether the ERK-dependent nuclear shuttling mechanisms previously characterized in Ref-1 cells (Le Gallic et al, 2004) are functional in C2C12 muscle cell lines. To test this, we performed subcellular fractionation experiments to determine the levels of ERF in the nuclear and cytoplasmic fractions of C2C12 muscle cell lines after incubation with U0126, an inhibitor for MEK, the upstream kinase for ERK1/2 at different times.…”

Section: Erf Can Physically Associate With the Utrophin-a N-box/ebs Sitementioning

confidence: 99%

“…MEK-dependent translocation of ERF from nucleus to cytoplasm was studied using C2C12 muscle cells that were incubated with MEK inhibitor U0126 for different times and analyzed using immunofluorescence of cells and immunoblotting of subcellular fractions as described previously for Ref1 cells (Le Gallic et al, 1999Gallic et al, , 2004. For immunofluorescent analysis, cells were fixed with ice-cold methanol for 10 min, incubated with anti-ERF antibodies (Santa Cruz Biotechnology), and detected using Alexa Fluor 546 donkey anti-goat secondary antibodies (Invitrogen, Carlsbad, CA).…”

Section: Tissue Culture and Transfectionmentioning

confidence: 99%

“…ERF is a novel member of the ets family and encodes a ubiquitously expressed 548-amino acid phospho-protein identified through its ability to repress the Ets-2 promoter via EBS binding (Sgouras et al, 1995;Le Gallic et al, 1999). ERF activity is regulated in part, by ERK-dependent changes in its phosphorylation status and subcellular localization due to nuclear shuttling, with CRM1 suggested to have a role in ERF export (Le Gallic et al, 2004). Here, we use an array of in vitro and in vivo methodologies to delineate the role of ERF in the regulation of the utrophin-A promoter.…”

mentioning

confidence: 99%

“…To demonstrate the existence of ERF and show its ability to form the complex in the context of muscle, we performed the gel shift and supershift by using nuclear extracts obtained from C2C12 muscle cell lines. As shown in Figure 1B, a protein-DNA complex was formed that was able to retard probe migration and able to be competed with excess unlabeled probe as well as supershifted using ERF-specific antibodies in muscle cell lines, suggesting the relevance of this mechanism for muscle in vivo.Having demonstrated the ability of ERF to associate with the N-box/EBS site of the utrophin-A promoter using gel shift assays, we asked whether the ERK-dependent nuclear shuttling mechanisms previously characterized in Ref-1 cells (Le Gallic et al, 2004) are functional in C2C12 muscle cell lines. To test this, we performed subcellular fractionation experiments to determine the levels of ERF in the nuclear and cytoplasmic fractions of C2C12 muscle cell lines after incubation with U0126, an inhibitor for MEK, the upstream kinase for ERK1/2 at different times.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Ets-2 Repressor Factor Silences Extrasynaptic Utrophin by N-Box–mediated Repression in Skeletal Muscle

Perkins

Basu

Budak

et al. 2007

MBoC

View full text Add to dashboard Cite

Utrophin is the autosomal homologue of dystrophin, the protein product of the Duchenne's muscular dystrophy (DMD) locus. Utrophin expression is temporally and spatially regulated being developmentally down-regulated perinatally and enriched at neuromuscular junctions (NMJs) in adult muscle. Synaptic localization of utrophin occurs in part by heregulin-mediated extracellular signal-regulated kinase (ERK)-phosphorylation, leading to binding of GABP␣/␤ to the N-box/EBS and activation of the major utrophin promoter-A expressed in myofibers. However, molecular mechanisms contributing to concurrent extrasynaptic silencing that must occur to achieve NMJ localization are unknown. We demonstrate that the Ets-2 repressor factor (ERF) represses extrasynaptic utrophin-A in muscle. Gel shift and chromatin immunoprecipitation studies demonstrated physical association of ERF with the utrophin-A promoter N-box/EBS site. ERF overexpression repressed utrophin-A promoter activity; conversely, small interfering RNA-mediated ERF knockdown enhanced promoter activity as well as endogenous utrophin mRNA levels in cultured muscle cells in vitro. Laser-capture microscopy of tibialis anterior NMJ and extrasynaptic transcriptomes and gene transfer studies provide spatial and direct evidence, respectively, for ERF-mediated utrophin repression in vivo. Together, these studies suggest "repressing repressors" as a potential strategy for achieving utrophin up-regulation in DMD, and they provide a model for utrophin-A regulation in muscle. INTRODUCTIONDuchenne's muscular dystrophy (DMD) is a fatal neuromuscular disease caused by gene mutations leading to qualitative or quantitative disturbances in dystrophin expression (Hoffman et al., 1987). Dystrophin-related protein or utrophin is considered the autosomal homologue of dystrophin, because it shares extensive sequence homology as well as its size and abundance in muscle (Love et al., 1989;Khurana et al., 1990;Tinsley et al., 1992). Utrophin and dystrophin also share functional properties such as the ability to associate with the dystroglycan complex and bind F-actin (Matsumura et al., 1992;Winder and Kendrick, 1995;Rybakova et al., 2002). Direct evidence for the ability of utrophin to functionally substitute comes from experiments demonstrating that utrophin overexpression driven either by transgenic means Rafael et al., 1998;Tinsley et al., 1998;Fisher et al., 2001) or viral vectors (Gilbert et al., 1999;Wakefield et al., 2000;Cerletti et al., 2003) can rescue dystrophin-deficient muscle. Despite these functional similarities, dystrophin and utrophin exhibit distinct localization in normal adult tissues.Perinatally, utrophin is expressed throughout the sarcolemma, however, its expression declines as that of dystrophin increases (Khurana et al., 1991;Clerk et al., 1993), leading to its spatially restricted expression at neuromuscular and myotendinous junctions (NMJs and MTJs, respectively) of adult myofibers. These features and relevance toward a therapy for DMD provide a powerful impetus for b...

show abstract

Noonan syndrome‐like phenotype associated with an ERF frameshift variant

Hirano,

Kuroda,

Enomoto

et al. 2024

American J of Med Genetics Pt A

View full text Add to dashboard Cite

Noonan syndrome is a so‐called “RASopathy,” that is characterized by short stature, distinctive facial features, congenital heart defects, and developmental delay. Of individuals with a clinical diagnosis of Noonan syndrome, 80%–90% have pathogenic variants in the known genes implicated in the disorder, but the molecular mechanism is unknown in the remaining cases. Heterozygous pathogenic variants of ETS2 repressor factor (ERF), which functions as a repressor in the RAS/MAPK signaling pathway, cause syndromic craniosynostosis. Here, we report an ERF frameshift variant cosegregating with a Noonan syndrome‐like phenotype in a family. The proband was a 3‐year‐old female who presented with dysmorphic facial features, including proptosis, hypertelorism, slightly down slanted palpebral fissures, low‐set posteriorly rotated ears, depressed nasal bridge, short stature, and developmental delay. Exome sequencing of the proband identified a heterozygous ERF variant [NM_006494.4: c.185del p.(Glu62Glyfs*15)]. Her mother and sister showed a similar phenotype and had the same heterozygous ERF variant. A large proportion of the previously reported patients with syndromic craniosynostosis and pathogenic ERF variants also showed characteristic features that overlap with those of Noonan syndrome. The present finding supports an association between heterozygous ERF variants and a Noonan syndrome‐like phenotype.

show abstract

ERF Nuclear Shuttling, a Continuous Monitor of Erk Activity That Links It to Cell Cycle Progression

Cited by 70 publications

References 61 publications

C-terminal domain small phosphatase 1 and MAP kinase reciprocally control REST stability and neuronal differentiation

C-terminal domain small phosphatase 1 and MAP kinase reciprocally control REST stability and neuronal differentiation

Ets-2 Repressor Factor Silences Extrasynaptic Utrophin by N-Box–mediated Repression in Skeletal Muscle

Noonan syndrome‐like phenotype associated with an ERF frameshift variant

Contact Info

Product

Resources

About