000030411 001__ 30411
000030411 005__ 20131002114013.0
000030411 0248_ $$aoai:doc.rero.ch:20121004154127-OZ$$punifr$$ppostprint$$prero_explore$$zcdu34$$zthesis_urn$$zcdu57$$zreport$$zthesis$$zbook$$zjournal$$zcdu16$$zpreprint$$zcdu1$$zdissertation
000030411 041__ $$aeng
000030411 080__ $$a57
000030411 100__ $$aPagac, Martin$$uDepartment of Biology, University of Fribourg, Switzerland - College of Pharmacy, University of Hawaii, Hilo, USA
000030411 245__ $$9eng$$aTopology of the microsomal glycerol-3-phosphate acyltransferase Gpt2p/Gat1p of Saccharomyces cerevisiae
000030411 269__ $$c2012-09-18
000030411 520__ $$9eng$$aAll glycerophospholipids are made from phosphatidic acid, which, according to the traditional view, is generated at the cytosolic surface of the ER. In yeast, phosphatidic acid is synthesized de novo by two acyl-CoA dependent acylation reactions. The first is catalyzed by one of the two homologous glycerol-3-phosphate acyltransferases Gpt2p/Gat1p and Sct1p/Gat2p, the second by one of the two 1-acyl-sn-glycerol-3-phosphate acyltransferases Slc1p and Ale1p/Slc4p. To study the biogenesis and topology of Gpt2p we observed the location of dual topology reporters inserted after various transmembrane helices. Moreover, using microsomes, we probed the accessibility of natural and substituted cysteine residues to a membrane impermeant alkylating agent and tested the protease sensitivity of various epitope tags inserted into Gpt2p. Finally, we assayed the sensitivity of the acyltransferase activity to membrane impermeant agents targeting lysine residues. By all these criteria we find that the most conserved motifs of Gpt2p and its functionally relevant lysines are oriented towards the ER lumen. Thus, the first step in biosynthesis of phosphatidic acid in yeast seems to occur in the ER lumen and substrates may have to cross the ER membrane.
000030411 695__ $$9eng$$aDual topology reporter ; cysteine accessibility ; phosphatidic acid ; lyso phosphatidic acid acyl Coenzyme A ; microsomes
000030411 700__ $$aVazquez, Hector M.$$uDepartment of Biology, University of Fribourg, Switzerland
000030411 700__ $$aBochud, Arlette$$uDepartment of Biology, University of Fribourg, Switzerland
000030411 700__ $$aRoubaty, Carole$$uDepartment of Biology, University of Fribourg, Switzerland
000030411 700__ $$aKnöpfli, Cécile$$uDepartment of Biology, University of Fribourg, Switzerland
000030411 700__ $$aVionnet, Christine$$uDepartment of Biology, University of Fribourg, Switzerland
000030411 700__ $$aConzelmann, Andreas$$uDepartment of Biology, University of Fribourg, Switzerland
000030411 773__ $$g2012///-$$tMolecular Biology
000030411 775__ $$gPublished version$$ohttp://dx.doi.org/10.1111/mmi.12047
000030411 8564_ $$fcon_tmg.pdf$$qapplication/pdf$$s642641$$uhttp://doc.rero.ch/record/30411/files/con_tmg.pdf$$yorder:1$$zpdf
000030411 8564_ $$fcon_tmg_sm.pdf$$qapplication/pdf$$s1581512$$uhttp://doc.rero.ch/record/30411/files/con_tmg_sm.pdf$$yorder:2$$zSupplementary material
000030411 918__ $$aFaculté des sciences$$bDécanat, Ch. du Musée 6A, 1700 Fribourg$$cBiologie
000030411 919__ $$aUniversité de Fribourg$$bFribourg$$ddoc.support@rero.ch
000030411 980__ $$aPOSTPRINT$$bUNIFR$$fART_JOURNAL
000030411 990__ $$a20121004154127-OZ