Open AccessDetermination of the membrane topology of Arv1 and the requirement of the ER luminal region for Arv1 function in Saccharomyces cerevisiae
Author(s) -
Villasmil Michelle L.,
Nickels, Jr Joseph T.
Publication year - 2011
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2011.00737.x
Subject(s) - saccharomyces cerevisiae , endoplasmic reticulum , membrane topology , biology , golgi apparatus , function (biology) , microbiology and biotechnology , transmembrane protein , membrane protein , membrane contact site , biochemistry , membrane , sphingolipid , topology (electrical circuits) , integral membrane protein , gene , receptor , mathematics , combinatorics
In Saccharomyces cerevisiae, ARV1 encodes a 321 amino acid transmembrane protein localized to the endoplasmic reticulum (ER) and Golgi. It has been shown previously that arv1 cells harbor defects in sphingolipid and glycosylphosphatidylinositol biosyntheses, and may harbor sterol trafficking defects. Using C‐terminal fusion to Suc2‐His4, we determined the orientation of full‐length Arv1 in the ER membrane. Once membrane topology was determined, we used this information and truncation analysis to establish the minimum protein length required for Arv1 function and phenotypic suppression. By understanding the topology of Arv1 we can now further analyze its putative lipid and glycosylphosphatidylinositol intermediate transport activities.