Differential targeting of MAP kinases to the ETS-domain transcription factor Elk-1

Yang, Shen Hsi

doi:10.1093/emboj/17.6.1740

Cited by 284 publications

(290 citation statements)

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“…A peptide derived from Elk1: QKGRKPRDLELPLSPSL (1934 Da) that contains a D-site (consensus underlined) (32), was synthesized at the UT Molecular Biology core facilities. The crude peptide was dissolved in equilibration buffer (0.1% trifluoroacetic acid) and purified by reverse phase chromatography using an Econosil C18 column (Alltech), developed with linear acetonitrile gradient 0-100% over 90 min using a flow rate 5 mL/min.…”

Section: Peptide Synthesis and Purificationmentioning

confidence: 99%

Expanding the Repertoire of an ERK2 Recruitment Site: Cysteine Footprinting Identifies the D-Recruitment Site as a Mediator of Ets-1 Binding

et al. 2007

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Many substrates of ERK2 contain a D-site, a sequence recognized by ERK2 that is used to promote catalysis. Despite lacking a canonical D-site, the substrate Ets-1 is displaced from ERK2 by peptides containing one. This suggests that Ets-1 may contain a novel or cryptic D-site. To investigate this possibility a protein footprinting strategy was developed to elucidate ERK2-ligand interactions. Using this approach, single cysteine reporters were placed in the D-recruitment site (DRS) of ERK2 and the resulting ERK2 proteins subjected to alkylation by iodoacetamide. The ability of residues 1-138 of Ets-1 to protect the cysteines from alkylation was determined. The pattern of protection observed is consistent with Ets-1 occupying a hydrophobic binding site within the DRS of ERK2. Significantly, a peptide derived from the D-site of Elk-1, which is known to bind the DRS, exhibits a similar pattern of cysteine protection. This analysis expands the repertoire of the DRS on ERK2 and suggests that other targeting sequences remain to be identified. Furthermore, cysteinefootprinting is presented as a useful way to interrogate protein-ligand interactions at the resolution of a single amino acid.The mitogen-activated protein kinases (MAPKs) 1 are ubiquitous elements of eukaryotic signaling pathways that permeate nearly all aspects of signaling in multicellular organisms. In humans the loss of their regulation is associated with many diseases that encompass an expanding list of cancers as well as neurological and inflammatory diseases (1). Three main subgroups have been identified in humans, termed the ERKs (2), the JNKs (3,4), and the p38 MAPKs (5,6). Several newer members have recently been reviewed (7). Extracellular regulated protein kinase 2 (ERK2), the prototypical member of the MAP kinase family, catalyzes the † This research was supported in part by the Welch Foundation (F-1390) and the National Institutes of Health (GM59802). Mass spectra were acquired by Dr. Herng-Hsiang Lo in the CRED Analytical Instrumentation Facility Core supported by the NIEHS center grant ES07784. Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081). NIH Public Access Author ManuscriptBiochemistry. Author manuscript; available in PMC 2010 July 6. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript transfer of the γ-phosphate of adenosine triphosphate to serine or threonine residues that generally lie in flexible regions of proteins. It is a remarkable enzyme whose ability to exhibit high specificity toward a discrete subset of structurally diverse proteins is poorly understood.Protein kinases generally recognize substrates through a consensus sequence that contains the phosphorylation site (8). This sequence is recognized, in part, by a region called the activation segment that encompasses residues 165 DFG to APE 195 of the catalytic domain of protein ki...

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Section: Peptide Synthesis and Purificationmentioning

confidence: 99%

Expanding the Repertoire of an ERK2 Recruitment Site: Cysteine Footprinting Identifies the D-Recruitment Site as a Mediator of Ets-1 Binding

et al. 2007

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“…Interaction of these docking sites with corresponding motifs in MAPKs could provide a molecular basis for stable and specific MAPK recruitment. A conserved MAPK domain, the common Journal of Cell Science 117 (17) Cano et al, 1995;Yang et al, 1998c;Yang et al, 1998b;Rao and Reddy, 1994 Yang et al, 1998b;Gupta et al, 1996 Jun GST pulldown, CIP Kallunki et al, 1996;Adler et al, 1992;Dai et al, 1995;Cano et al, 1995;Read et al, 1997;Gupta et al, 1996;Kallunki et al, 1994;Sluss et al, 1994;Hibi et al, 1993;Derijard et al, 1994 SAP-2 GST pulldown Ducret et al, 2000 ATF-2 GST pulldown Livingstone et al, 1995;Gupta et al, 1995;Gupta et al, 1996;Raingeaud et al, 1995 JunB GST pulldown Gupta et al, 1996;Kallunki et al, 1996 docking (CD) domain, is often used in the interaction with docking sites in MAPKKs, MKPs (MAPK-phosphatases) and MAPKAPKs (Tanoue and Nishida, 2003). However, whether or not the D domain of particular transcription factors can interact with the CD domain is not known.…”

Section: Protein Domains Potentially Involved In Stable and Specific mentioning

confidence: 99%

MAP kinases as structural adaptors and enzymatic activators in transcription complexes

Edmunds

Mahadevan

2004

Journal of Cell Science

103

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Mitogen-activated protein kinase (MAPK) pathways regulate eukaryotic gene expression in response to extracellular stimuli. MAPKs and their downstream kinases phosphorylate transcription factors, co-regulators and chromatin proteins to initiate transcriptional changes. However, the spatial context in which the MAPKs operate in transcription complexes is poorly understood. Recent findings in budding yeast show that MAPKs can form integral components of transcription complexes and have novel structural functions in addition to phosphorylating local substrates. Hog1p MAPK is stably recruited to target promoters by specific transcription factors in response to osmotic stress, and acts as both a structural adaptor and enzymatic activator driving the assembly and activation of the transcription complex. We review the evidence that suggests a similar bifunctional role for MAPKs in mammalian transcription complexes.

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“…KIMs come in different variations that can interact with any of the three MAPKs or only a subset [151,153,154]. A meticulous mutational dissection of the KIM in the transcription factor Elk has unveiled subtle differences between ERK and JNK binding to the KIM, and shown that p38 does not require the KIM at all to phosphorylate Elk [155]. However, sequence comparisons between different KIMs do not disclose designated binding motifs for specific MAPKs (Figure 7).…”

Section: Picking Activators and Substrates : Erk Docking Sitesmentioning

confidence: 99%

Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions

Kölch

2000

Biochemical Journal

1,057

118

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The Ras\Raf\MEK (mitogen-activated protein kinase\ERK kinase)\ERK (extracellular-signal-regulated kinase) pathway is at the heart of signalling networks that govern proliferation, differentiation and cell survival. Although the basic regulatory steps have been elucidated, many features of this pathway are only beginning to emerge. This review focuses on the role of protein-protein interactions in the regulation of this pathway, INTRODUCTIONThe mitogen-activated protein kinase (MAPK) pathway is one of the primordial signalling systems that Nature has used in several permutations to accomplish an amazing variety of tasks. It exists in all eukaryotes, and controls such fundamental cellular processes as proliferation, differentiation, survival and apoptosis. The basic arrangement includes a G-protein working upstream of a core module consisting of three kinases : a MAPK kinase kinase (MAPKKK) that phosphorylates and activates a MAPK kinase (MAPKK), which in turn activates MAPK ( Figure 1). This set-up provides not only for signal amplification, but, maybe even more importantly, for additional regulatory interfaces that allow the kinetics, duration and amplitude of the activity to be precisely tuned. At present we can distinguish six MAPK modules, which share structurally related components, but seem to mediate specific biological responses. For recent reviews on MAPK pathways, the reader is referred to references [1][2][3]. Here we will focus on the regulation of the ERK (extracellular-signal-regulated kinase) pathway, which features Ras as G-protein, Raf as MAPKKK, MEK (MAPK\ERK kinase) as MAPKK and ERK as MAPK.Despite enjoying a decade in the limelight of scientific interest and revealing a plethora of new insights into the circuitry of signalling pathways in general, this pathway still holds many secrets. These pertain mainly to the regulation of Raf, and to a deeper understanding of how specific biological responses are encoded by spatial and temporal changes in the activity and subcellular distribution of the pathway components, and how these fluctuations are orchestrated at the molecular level. Work from many laboratories has highlighted protein-protein interactions as powerful means of co-ordinating signalling processes, most strikingly exemplified by the assembly of multi-protein signalling complexes on activated receptors, or of transcriptionfactor complexes on gene promoters. However, it is becoming Abbreviations used : Bcr, Breakpoint cluster region ; BXB, isolated Raf-1 kinase domain ; CNK, connector-enhancer of KSR ; CK2, casein kinase 2 ; CRD, cysteine-rich domain ; DEF, docking site for ERK, FxFP ; DLK, dual leucine-zipper-bearing kinase ; ERK, extracellular-signal-regulated kinase ; Hsp, heat-shock protein ; KIM, kinase interaction motif ; IκB, inhibitor of NF-κB ; JNK, c-Jun N-terminal kinase ; KSR, kinase suppressor of Ras ; MAPK, mitogen-activated protein kinase ; MAPKK, MAPK kinase ; MAPKKK, MAPK kinase kinase ; MEK, MAPK/ERK kinase ; MEKK, MEK kinase ; MP1, MEK partner 1 ; MUK, MAPK upstream kin...

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Differential targeting of MAP kinases to the ETS-domain transcription factor Elk-1

Cited by 284 publications

References 42 publications

Expanding the Repertoire of an ERK2 Recruitment Site: Cysteine Footprinting Identifies the D-Recruitment Site as a Mediator of Ets-1 Binding

Expanding the Repertoire of an ERK2 Recruitment Site: Cysteine Footprinting Identifies the D-Recruitment Site as a Mediator of Ets-1 Binding

MAP kinases as structural adaptors and enzymatic activators in transcription complexes

Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions

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