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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.
J Biol Chem. 2016 Feb 12; 291(7):3124-34.JB

Abstract

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 α-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.

Authors+Show Affiliations

From the Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.the Department of Human Genetics, Catholic University of Leuven, 3000 Leuven, Belgium.From the Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.From the Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.From the Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.From the Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.the Department of Genetics, Hospital Universitario Cruces, BioCruces Health Research Institute, Biscay, Spain.Department of Pharmaceutical Sciences, Hokkaido University, N12W6, Sapporo 060-0812, Japan, and.the Medical Genomics Laboratory, Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294.the Department of Human Genetics, Catholic University of Leuven, 3000 Leuven, Belgium.From the Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan, yoshimura@a6.keio.jp.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26635368

Citation

Hirata, Yasuko, et al. "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 Journal of Biological Chemistry, vol. 291, no. 7, 2016, pp. 3124-34.
Hirata Y, Brems H, Suzuki M, et al. 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. J Biol Chem. 2016;291(7):3124-34.
Hirata, Y., Brems, H., Suzuki, M., Kanamori, M., Okada, M., Morita, R., Llano-Rivas, I., Ose, T., Messiaen, L., Legius, E., & Yoshimura, A. (2016). 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 Journal of Biological Chemistry, 291(7), 3124-34. https://doi.org/10.1074/jbc.M115.703710
Hirata Y, et al. 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. J Biol Chem. 2016 Feb 12;291(7):3124-34. PubMed PMID: 26635368.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Hirata,Yasuko, AU - Brems,Hilde, AU - Suzuki,Mayu, AU - Kanamori,Mitsuhiro, AU - Okada,Masahiro, AU - Morita,Rimpei, AU - Llano-Rivas,Isabel, AU - Ose,Toyoyuki, AU - Messiaen,Ludwine, AU - Legius,Eric, AU - Yoshimura,Akihiko, Y1 - 2015/12/03/ PY - 2015/11/11/received PY - 2015/12/5/entrez PY - 2015/12/5/pubmed PY - 2016/7/7/medline KW - GTPase-activating protein (GAP) KW - Ras protein KW - human genetics KW - mitogen-activated protein kinase (MAPK) KW - negative regulation KW - protein domain KW - signal transduction SP - 3124 EP - 34 JF - The Journal of biological chemistry JO - J Biol Chem VL - 291 IS - 7 N2 - 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 α-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. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/26635368/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_ L2 - https://linkinghub.elsevier.com/retrieve/pii/M115.703710 DB - PRIME DP - Unbound Medicine ER -