Septin higher-order structure on yeast membranes in vitro.

TitleSeptin higher-order structure on yeast membranes in vitro.
Publication TypeJournal Article
Year of Publication2025
AuthorsGoodchild JA, Curtis BN, Pan Y, Jiang Y, Jiao F, Gladfelter AS, Scheuring S
JournalNat Commun
Volume16
Issue1
Pagination5055
Date Published2025 May 30
ISSN2041-1723
KeywordsCell Membrane, Cytoskeleton, Lipid Bilayers, Microscopy, Atomic Force, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Septins
Abstract

Septins are cytoskeletal proteins that form filaments and higher-order structures, and remodel membranes in a variety of processes. Structural and cell biological studies provided atomic- and micro-scale details, but the understanding of septin assembly at the mesoscale is limited. Here, we used high-speed atomic force microscopy (HS-AFM) to analyze yeast septin assembly on yeast supported lipid bilayers (SLBs). We found the coexistence of three lipid phases in yeast membranes, where septin polymerized selectively on the liquid-disordered phase. Septin filaments adhered to membranes with a conserved face; and paired filaments, previously reported in less native environments, were not observed. Additionally, septin filaments exhibited lateral and longitudinal alignment. We used HS-AFM force-sweep experiments to disrupt septin structures and observe organizational recovery through self-templating. Finally, septin filaments stacked, where higher layer filament alignment was templated by the layer below. Thus, septins encode their 3D-structural organization, likely tunable by the membrane and bulk environment.

DOI10.1038/s41467-025-60344-w
Alternate JournalNat Commun
PubMed ID40447632
PubMed Central IDPMC12125352
Grant ListR01GM130934 / / U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS) /
DP1AT010874 / / U.S. Department of Health & Human Services | NIH | National Center for Complementary and Integrative Health (NCCIH) /
R01 NS134559 / NS / NINDS NIH HHS / United States
R01NS134559 / / U.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and Stroke (NINDS) /
DP1 AT010874 / AT / NCCIH NIH HHS / United States
R01 GM130934 / GM / NIGMS NIH HHS / United States