title: Tyr728 in the kinase domain of the murine kinase suppressor of RAS 1 regulates binding and activation of the mitogen-activated protein kinase kinase.
abstract: In metazoans, the highly conserved MAPK signaling pathway regulates cell fate decision. Aberrant activation of this pathway has been implicated in multiple human cancers and some developmental disorders. KSR1 functions as an essential scaffold that binds the individual components of the cascade and coordinates their assembly into multiprotein signaling platforms. The mechanism of KSR1 regulation is highly complex and not completely understood. In this study, we identified Tyr(728) as a novel regulatory phosphorylation site in KSR1. We show that Tyr(728) is phosphorylated by LCK, uncovering an additional and unexpected link between Src kinases and MAPK signaling. To understand how phosphorylation of Tyr(728) may regulate the role of KSR1 in signal transduction, we integrated structural modeling and biochemical studies. We demonstrate that Tyr(728) is involved in maintaining the conformation of the KSR1 kinase domain required for binding to MEK. It also affects phosphorylation and activation of MEK by RAF kinases and consequently influences cell proliferation. Moreover, our studies suggest that phosphorylation of Tyr(728) may affect the intrinsic kinase activity of KSR1. Together, we propose that phosphorylation of Tyr(728) may regulate the transition between the scaffolding and the catalytic function of KSR1 serving as a control point used to fine-tune cellular responses.
authors.affiliation: From the Department of Microbiology, Biocenter, and.
authors.forename: Claudia
authors.initials: C
authors.lastname: Sibilski
authors.forename: Thomas
authors.initials: T
authors.lastname: Mueller
authors.forename: Laxmikanth
authors.initials: L
authors.lastname: Kollipara
authors.forename: René P
authors.initials: RP
authors.lastname: Zahedi
authors.forename: Ulf R
authors.initials: UR
authors.lastname: Rapp
authors.forename: Thomas
authors.initials: T
authors.lastname: Rudel
authors.forename: Angela
authors.initials: A
authors.lastname: Baljuls
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D004789:Enzyme Activation; D006801:Humans; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism; D020935:MAP Kinase Signaling System; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism*; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014443:Tyrosine / Q000737:chemistry
chemical_list: D014443:Tyrosine; D011494:Protein Kinases; C097199:KSR-1 protein kinase; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D020929:Mitogen-Activated Protein Kinase Kinases
keywords: KSR1; MAP Kinases (MAPKs); Mass Spectrometry (MS); Molecular Modeling; Raf; Signal Transduction; Src Family Kinases; Tyrosine Phosphorylation