Open AccessTargeting and membrane insertion into the endoplasmic reticulum membrane of Saccharomyces cerevisiae essential protein Rot1
Author(s) -
Juanes María Angeles,
MartínezGaray Carlos Andrés,
Igual Juan Carlos,
Bañó María Carmen
Publication year - 2010
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.1567-1364.2010.00653.x
Subject(s) - endoplasmic reticulum , biology , saccharomyces cerevisiae , membrane contact site , membrane protein , membrane , microbiology and biotechnology , stim1 , biochemistry , integral membrane protein , yeast
Rot1 is an essential yeast protein that has been related to cell wall biosynthesis, actin cytoskeleton dynamics and protein folding. Rot1 is an N ‐glycosylated protein anchored to the nuclear envelope–endoplasmic reticulum (ER) membrane by a transmembrane domain at its C‐terminal end. Rot1 is translocated to the ER by a post‐translational mechanism. Here, we investigate the protein domain required to target and translocate Rot1 to the ER membrane. We found that several deletions of the N‐terminal region of Rot1 prevented neither membrane targeting nor the insertion of this protein. Interestingly, we obtained the same results when different truncated forms in the C‐terminal transmembrane domain were analyzed, suggesting the presence of an internal topogenic element that is capable of translocating Rot1 to the ER. To identify this sequence, we generated a combination of N‐ and C‐terminal deletion mutants of Rot1 and we investigated their insertion into the membrane. The results show that two regions, amino acids 26–60 and 200–228, are involved in the post‐translational translocation of Rot1 across the ER membrane.