Localization of Gts1p in cortical actin patches of yeast and its possible role in endocytosis.

Herein we report that Gts1p fused with green-fluorescent protein (GFP) is localized in the cortical actin patch besides nuclei in yeast and the cortical Gts1p changed its position together with the patch depending on the cell-cycle phase, while nuclear Gts1p accumulated predominantly in the budding phase. Whereas Gts1p does not ...
directly bind to actin, it associated mainly with the actin-associated protein Pan1p. In the GTS1-deleted transformant gts1Delta, the number of cells containing either a fragmented vacuole or an enlarged single central vacuole increased and the uptake of the hydrophilic dye Lucifer yellow (LY) in the vacuole decreased. Further, gts1Delta transformed with a mutant Gts1p having two cysteine-to-alanine substitutions in a zinc finger resembling that of GTPase-activating proteins of ADP-ribosylation factors (ARF-GAP) neither recovered the LY uptake unlike gts1Delta transformed with the wild-type GTS1, nor reduced the average size of central vacuoles as much as the latter did. These results suggested that Gts1p in the actin patch is involved in the fluid-phase endocytosis and membrane trafficking for vacuole formation and that the putative ARF-GAP domain in Gts1p plays an important role in these functions.
Mesh Terms:
ADP-Ribosylation Factors, Actins, Amino Acid Sequence, Bicyclo Compounds, Heterocyclic, Endocytosis, GTPase-Activating Proteins, Gene Deletion, Green Fluorescent Proteins, Immunoprecipitation, Isoquinolines, Molecular Sequence Data, Protein Structure, Tertiary, Protein Transport, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Subcellular Fractions, Thiazolidines, Transcription Factors, Vacuoles
Eur. J. Cell Biol.
Date: May. 01, 2007
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