Basic clusters and amphipathic helices contribute to interactions of Myr1/Syh1 with membrane phospholipids
(English)Manuscript (preprint) (Other academic)
The ability to associate transiently with membrane bilayers is an important property of many protein regulators of membrane trafficking, lipid transfer proteins, or signaling modules. Membrane association is also a property of Myr1/Syh1, a soluble GYF domain protein from Saccharomyces cerevisiae, previously reported to rescue the temperature sensitive growth of ypt6 and ric1 null strains. Here, we further demonstrate that MYR1 also rescued the vacuole fragmentation phenotype of the ypt6 and ric1 mutants. The mechanism behind these genetic interactions is likely linked to the capacity of the Myr1/Syh1 protein to associate with phospholipid membranes. In order to elucidate further the nature of the interactions with vesicular traffic, we studied protein-protein and protein-phospholipid association of isolated domains from Myr1/Syh1. Using a two-hybrid assay, we confirmed the capacity of Myr1/Syh1 to self-associate in vivo. We measured in vitro the affinity of recombinant Myr1/Syh1 domains fused to GFP for liposomes reconstituted from synthetic and natural yeast lipids by sedimentation techniques. The herewith established affinities of Myr1/Syh1 to specific lipids, combined with evidence for its interactions with membrane traffic and protein synthesis, provide support for a possible function of Myr1/Syh1 as a regulator sensing membrane composition along the vesicular pathways.
Biochemistry and Molecular Biology
Research subject Biochemistry
IdentifiersURN: urn:nbn:se:su:diva-62030OAI: oai:DiVA.org:su-62030DiVA: diva2:439464