000199658 001__ 199658
000199658 005__ 20181203023520.0
000199658 0247_ $$2doi$$a10.1074/jbc.M113.537001
000199658 022__ $$a0021-9258
000199658 02470 $$2ISI$$a000333807000009
000199658 037__ $$aARTICLE
000199658 245__ $$aThe Efficacy of Raf Kinase Recruitment to the GTPase H-ras Depends on H-ras Membrane Conformer-specific Nanoclustering
000199658 260__ $$bAmerican Society for Biochemistry and Molecular Biology$$c2014$$aBethesda
000199658 269__ $$a2014
000199658 300__ $$a15
000199658 336__ $$aJournal Articles
000199658 520__ $$aBackground: Ras nanoclusters contain 6-8 Ras proteins on the plasma membrane and serve as indispensable signaling platforms for Ras-MAPK signaling. Results: Ras membrane conformer mutants impart specific galectin-1-dependent nanoclustering responses. Conclusion: Mutations in Ras can affect its nanoclustering response and thus allosterically effector recruitment and downstream signaling. Significance: Disease-associated mutations that perturb Ras membrane conformers may alter signaling through nanoclustering. Solution structures and biochemical data have provided a wealth of mechanistic insight into Ras GTPases. However, information on how much the membrane organization of these lipid-modified proteins impacts on their signaling is still scarce. Ras proteins are organized into membrane nanoclusters, which are necessary for Ras-MAPK signaling. Using quantitative conventional and super-resolution fluorescence methods, as well as mathematical modeling, we investigated nanoclustering of H-ras helix 4 and hypervariable region mutants that have different bona fide conformations on the membrane. By following the emergence of conformer-specific nanoclusters in the plasma membrane of mammalian cells, we found that conformers impart distinct nanoclustering responses depending on the cytoplasmic levels of the nanocluster scaffold galectin-1. Computational modeling revealed that complexes containing H-ras conformers and galectin-1 affect both the number and lifetime of nanoclusters and thus determine the specific Raf effector recruitment. Our results show that mutations in Ras can affect its nanoclustering response and thus allosterically effector recruitment and downstream signaling. We postulate that cancer- and developmental disease-linked mutations that are associated with the Ras membrane conformation may exhibit so far unrecognized Ras nanoclustering and therefore signaling alterations.
000199658 6531_ $$aCancer
000199658 6531_ $$aFluorescence Correlation Spectroscopy
000199658 6531_ $$aFluorescence Resonance Energy Transfer (FRET)
000199658 6531_ $$aRas
000199658 6531_ $$aSignaling
000199658 6531_ $$aFluorescence Recovery after Photobleaching (FRAP)
000199658 6531_ $$aFluorescence-lifetime Microscopy Imaging (FLIM)
000199658 6531_ $$aStimulated Emission Depletion (STED)
000199658 6531_ $$aMembrane
000199658 6531_ $$aNanocluster
000199658 700__ $$uAbo Akad Univ, Turku Ctr Biotechnol, FIN-20520 Turku, Finland$$aGuzman, Camilo
000199658 700__ $$uAbo Akad Univ, Turku Ctr Biotechnol, FIN-20520 Turku, Finland$$aSolman, Maja
000199658 700__ $$uAbo Akad Univ, Turku Ctr Biotechnol, FIN-20520 Turku, Finland$$aLigabue, Alessio
000199658 700__ $$uAbo Akad Univ, Turku Ctr Biotechnol, FIN-20520 Turku, Finland$$aBlazevits, Olga
000199658 700__ $$uMax Planck Inst Biophys Chem, Dept Nanobiophoton, D-37077 Gottingen, Germany$$aAndrade, Debora M.
000199658 700__ $$0240935$$g155775$$uEcole Polytech Fed Lausanne, Inst Chem Sci & Engn, CH-1015 Lausanne, Switzerland$$aReymond, Luc
000199658 700__ $$uMax Planck Inst Biophys Chem, Dept Nanobiophoton, D-37077 Gottingen, Germany$$aEggeling, Christian
000199658 700__ $$aAbankwa, Daniel$$uAbo Akad Univ, Turku Ctr Biotechnol, FIN-20520 Turku, Finland
000199658 773__ $$j289$$tJournal Of Biological Chemistry$$k14$$q9519-9533
000199658 909C0 $$xU10102$$0252027$$pLIP
000199658 909CO $$pSB$$particle$$ooai:infoscience.tind.io:199658
000199658 917Z8 $$x105423
000199658 917Z8 $$x148230
000199658 917Z8 $$x148230
000199658 937__ $$aEPFL-ARTICLE-199658
000199658 973__ $$rREVIEWED$$sPUBLISHED$$aOTHER
000199658 980__ $$aARTICLE