Przejdź do zawartości
Merck

Synergistic role of nucleotides and lipids for the self-assembly of Shs1 septin oligomers.

The Biochemical journal (2020-07-30)
Cyntia Taveneau, Rémi Blanc, Gérard Péhau-Arnaudet, Aurélie Di Cicco, Aurélie Bertin
ABSTRAKT

Budding yeast septins are essential for cell division and polarity. Septins assemble as palindromic linear octameric complexes. The function and ultra-structural organization of septins are finely governed by their molecular polymorphism. In particular, in budding yeast, the end subunit can stand either as Shs1 or Cdc11. We have dissected, here, for the first time, the behavior of the Shs1 protomer bound to membranes at nanometer resolution, in complex with the other septins. Using electron microscopy, we have shown that on membranes, Shs1 protomers self-assemble into rings, bundles, filaments or two-dimensional gauzes. Using a set of specific mutants we have demonstrated a synergistic role of both nucleotides and lipids for the organization and oligomerization of budding yeast septins. Besides, cryo-electron tomography assays show that vesicles are deformed by the interaction between Shs1 oligomers and lipids. The Shs1-Shs1 interface is stabilized by the presence of phosphoinositides, allowing the visualization of micrometric long filaments formed by Shs1 protomers. In addition, molecular modeling experiments have revealed a potential molecular mechanism regarding the selectivity of septin subunits for phosphoinositide lipids.

MATERIAŁY
Numer produktu
Marka
Opis produktu

Sigma-Aldrich
L-α-Phosphatidylethanolamine, dioleoyl, ≥99% (GC), ≥98% (TLC), lyophilized powder
Sigma-Aldrich
Guanosine 5′-triphosphate lithium salt, ~95% (HPLC), powder
Sigma-Aldrich
Guanosine 5′-diphosphate sodium salt, Type I, ≥96% (HPLC)
Millipore
Benzonase® Nuclease, ≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
Sigma-Aldrich
1,2-Dioleoyl-sn-glycero-3-phosphocholine, lyophilized powder