1.15 Å Crystal structure of the <i>X. tropicalis</i> Spred1 EVH1 domain suggests a fourth distinct peptide‐binding mechanism within the EVH1 family

Harmer, Nicholas J.; Sivak, Jeremy M.; Amaya, Enrique; Blundell, Tom L.

doi:10.1016/j.febslet.2004.11.114

Cited by 18 publications

(23 citation statements)

References 31 publications

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“…The crystal structure of the Xenopus tropicalis Spred1 EVH1 domain suggests a distinct peptide-binding mechanism compared with other identified EVH1 domains (Harmer et al 2005). Although an interacting protein for the Spred2 EVH1 domain has recently been reported (NBR1) (Mardakheh et al 2009), proteins that specifically interact with the Spred1 EVH1 domain have yet to be determined.…”

Section: Resultsmentioning

confidence: 99%

A shared molecular mechanism underlies the human rasopathies Legius syndrome and Neurofibromatosis-1

Stowe

Mercado²,

Stowe³

et al. 2012

Genes Dev.

130

148

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The Ras/mitogen-activated protein kinase (MAPK) pathway plays a critical role in transducing mitogenic signals from receptor tyrosine kinases. Loss-of-function mutations in one feedback regulator of Ras/MAPK signaling, SPRED1 (Sprouty-related protein with an EVH1 domain), cause Legius syndrome, an autosomal dominant human disorder that resembles Neurofibromatosis-1 (NF1). Spred1 functions as a negative regulator of the Ras/MAPK pathway; however, the underlying molecular mechanism is poorly understood. Here we show that neurofibromin, the NF1 gene product, is a Spred1-interacting protein that is necessary for Spred1's inhibitory function. We show that Spred1 binding induces the plasma membrane localization of NF1, which subsequently down-regulates Ras-GTP levels. This novel mechanism for the regulation of neurofibromin provides a molecular bridge for understanding the overlapping pathophysiology of NF1 and Legius syndrome.

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Section: Resultsmentioning

confidence: 99%

A shared molecular mechanism underlies the human rasopathies Legius syndrome and Neurofibromatosis-1

Stowe

Mercado²,

Stowe³

et al. 2012

Genes Dev.

130

148

View full text Add to dashboard Cite

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“…The SPRED1 EVH-l domain has been suggested to bind to a ligand in a groove formed by ␤-strands ␤l, ␤2, ␤6, and ␤7 (38). Missense mutations with a severe effect on GRD binding such as G30R, W31C/W31L, V44D, G62R, G100D, and T102K/ T102R are probably present in the groove for the binding to the GRD or important for the structure of the binding groove.…”

Section: Evh1 Missense Mutations Found In Legius Syndromementioning

confidence: 99%

Interaction between a Domain of the Negative Regulator of the Ras-ERK Pathway, SPRED1 Protein, and the GTPase-activating Protein-related Domain of Neurofibromin Is Implicated in Legius Syndrome and Neurofibromatosis Type 1

Hirata

Brems

Suzuki

et al. 2016

Journal of Biological Chemistry

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Constitutional heterozygous loss-of-function mutations in the SPRED1 gene cause a phenotype known as Legius syndrome, which consists of symptoms of multiple café-au-lait macules, axillary freckling, learning disabilities, and macrocephaly. Legius syndrome resembles a mild neurofibromatosis type 1 (NF1) phenotype. It has been demonstrated that SPRED1 functions as a negative regulator of the Ras-ERK pathway and interacts with neurofibromin, the NF1 gene product. However, the molecular details of this interaction and the effects of the mutations identified in Legius syndrome and NF1 on this interaction have not yet been investigated. In this study, using a yeast two-hybrid system and an immunoprecipitation assay in HEK293 cells, we found that the SPRED1 EVH1 domain interacts with the N-terminal 16 amino acids and the C-terminal 20 amino acids of the GTPase-activating protein (GAP)-related domain (GRD) of neurofibromin, which form two crossing ␣-helix coils outside the GAP domain. These regions have been shown to be dispensable for GAP activity and are not present in p120 GAP . Several mutations in these N-and C-terminal regions of the GRD in NF1 patients and pathogenic missense mutations in the EVH1 domain of SPRED1 in Legius syndrome reduced the binding affinity between the EVH1 domain and the GRD. EVH1 domain mutations with reduced binding to the GRD also disrupted the ERK suppression activity of SPRED1. These data clearly demonstrate that SPRED1 inhibits the Ras-ERK pathway by recruiting neurofibromin to Ras through the EVH1-GRD interaction, and this study also provides molecular basis for the pathogenic mutations of NF1 and Legius syndrome.Spred (Sprouty-related protein with an EVH1 domain) family proteins, initially discovered as c-Kit-and c-Fms-binding proteins, have been shown to suppress the Ras-ERK pathway. Spreds form a subfamily of the Sprouty/Spred family, which is characterized by the Sprouty-related C-terminal cysteine-rich (SPR) 3 domain. In mammals, four Sprouty homologues and three members of the Spred family of proteins, Spred1, Spred2, and Spred3, have been discovered. Mammalian Spreds can negatively regulate the Ras-Raf-ERK pathway (1, 2); however, compared with Spred1 and Spred2, Spred3 has much weaker ERK suppression activity (3). The SPR domain has been shown to be palmitoylated, causing Sprouty and Spred to localize in the membrane fraction (4, 5). Spred1 and Spred2 are composed of an N-terminal enabled/vasodilator-stimulated protein homology 1 (EVH1) domain, a central c-Kit-binding domain, and a cysteine-rich C-terminal SPR domain, whereas Spred3 lacks a functional c-Kit-binding domain.

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“…Recently, structural information on the Spred EVH1 domain revealed that this domain adopts, like the other EVH1 domains, a pleckstrin homology fold (48,49). However, for the Spred EVH1 domain a different induced fit peptide binding mechanism has been proposed even in the absence of a characterized ligand for this domain (48,49), which makes the identification of binding partners a point of major interest (Fig. 1).…”

Section: Domains: Structure and Functionsmentioning

confidence: 99%

The VASP-Spred-Sprouty Domain Puzzle

Bundschu

Walter

Schuh

2006

Journal of Biological Chemistry

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Sprouty-related proteins with an EVH1 domain (Spreds) belong to a new protein family harboring a conserved N-terminal EVH1 domain, which is related to the VASP (vasodilatorstimulated phosphoprotein) EVH1 domain (Enabled/VASP homology 1 domain) and a C-terminal Sprouty-related domain, typical for Sprouty proteins. Spreds were, like Sproutys, initially discovered as inhibitors of the Ras/MAPK pathway, and the SPR (Sprouty-related) domains of both protein families seem to be very important for many protein interactions and cellular processes. VASP was initially characterized as a proline-rich substrate of protein kinases A and G in human platelets and later shown to be a scaffold protein, regulating both signal transduction pathways and the actin filament system. The VASP-EVH1 domain is known to bind specifically to a FP 4 binding motif, which is, for example, present in the focal adhesion proteins vinculin and zyxin. In this review we give a structural and functional overview on these three protein families and ask whether nature plays a modular protein domain puzzle with stable exchangeable elements or if these closely related domains have various functions when pasted in a different protein context. Family Members, Structure, and ExpressionSpred-Spred 2 proteins are a new family of membrane-associated suppressors of growth factor-induced ERK activation. The Drosophila founder member AE33 (1) was followed by human, mouse, and Xenopus Spreds (2, 3) and, recently, by Eve-3, a splice variant of Spred-3 (4). Spred family members contain an N-terminal EVH1 domain, a central c-Kit binding domain (KBD), and a C-terminal Sprouty-like cysteine-rich domain (SPR domain) (2, 5). Spred-3 lacks a functional c-Kit binding domain (2), and Eve-3 consists of merely a single EVH1 domain (4). So far, four binding partners were described, namely Raf1, caveolin-1, Ras, and RhoA (5-8). The former two bind to the Spred-SPR domain, but for the latter two no binding sites are known so far. Spred-1 is expressed predominantly in adult brain (9) and in some fetal tissues, suggesting a role during development (9). Spred-2 expression was rare in fetal tissues but mostly ubiquitous in adults, whereas an especially strong expression was seen in neural tissues and different glandular epithelia (9, 10). Spred-3 was detected only in brain (2, 9), whereas Eve-3 was limited to the developed liver (4).VASP-Ena/VASP (vasodilator-stimulated phosphoprotein) proteins contain, like Spreds, an N-terminal EVH1 domain. Mammalian VASP was first characterized in platelets as a proline-rich substrate of cAMP-and cGMP-dependent protein kinases (11). Additional members of the Ena/VASP family are the Drosophila Enabled (Ena), its mammalian homologue Mena (mammalian Enabled), the C. elegans homologue unc34, and the murine Evl (Enabled/vasodilator-stimulated phosphoprotein-like) (12)(13)(14). In general, the EVH1 domain is followed by a central proline-rich region responsible for interaction with Src homology 3 domains and profilins (compiled in Ref. 15) and, finally...

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1.15 Å Crystal structure of the X. tropicalis Spred1 EVH1 domain suggests a fourth distinct peptide‐binding mechanism within the EVH1 family

Cited by 18 publications

References 31 publications

A shared molecular mechanism underlies the human rasopathies Legius syndrome and Neurofibromatosis-1

A shared molecular mechanism underlies the human rasopathies Legius syndrome and Neurofibromatosis-1

Interaction between a Domain of the Negative Regulator of the Ras-ERK Pathway, SPRED1 Protein, and the GTPase-activating Protein-related Domain of Neurofibromin Is Implicated in Legius Syndrome and Neurofibromatosis Type 1

The VASP-Spred-Sprouty Domain Puzzle

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