Correlation analyses reveal differential diffusion behavior of eisosomal proteins between mother and daughter cells

Methods Appl Fluoresc. 2022 Sep 14;10(4). doi: 10.1088/2050-6120/ac8fe1.

Abstract

Eisosomes are nanoscale plasma membrane domains shaped as furrow-like invaginations. InSaccharomyces cerevisiaethese relatively immobile and uniform structures are mainly composed of two cytoplasmic proteins Pil1 and Lsp1. The present work uses fluctuation of fluorescence signals and analytical methods to determine Pil1 and Lsp1 dynamics at different subcellular locations. Using scanning techniques and autocorrelation analysis we determine that the cytoplasmic pools of Pil1 and Lsp1 behave mainly by passive diffusion. Single-point FCS experiments performed at several subcellular locations reveal that Pil1 mobility is faster in daughter cells. Furthermore, pair correlation function analysis indicates a rapid dynamic of Pil1 near the plasma membrane of growing yeast buds, where the membrane is expected to be actively assembling eisosomes.

Keywords: correlation techniques; eisosomes; fluctuation analyses; intracellular dynamics.

MeSH terms

  • Cell Membrane / metabolism
  • Female
  • Humans
  • Mothers
  • Phosphoproteins / chemistry
  • Phosphoproteins / metabolism
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins* / chemistry
  • Saccharomyces cerevisiae Proteins* / metabolism

Substances

  • Phosphoproteins
  • Saccharomyces cerevisiae Proteins