NDLI: Functional expression of human mdr1 in the yeast Saccharomyces cerevisiae.

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Functional expression of human mdr1 in the yeast Saccharomyces cerevisiae.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Thorner, J. Kuchler, K.
Description	Author Affiliation: Kuchler K ( Department of Molecular and Cell Biology, University of California, Berkeley 94720.);
Abstract	Development of multiple drug resistance in tumor cells involves amplification of the mdr1 gene product, a 170-kDa plasma membrane glycoprotein that is an ATP-driven pump that extrudes the drugs. Human mdr1 (also designated as PGY1) cDNA was expressed in yeast cells by using the promoter and translational initiation signal of a related yeast gene, STE6. Immunoblotting of subcellular fractions showed that all of the Mdr1 (also known as P glycoprotein) was associated with the particulate material. Immunofluorescence microscopy revealed that the majority of the Mdr1 was localized to the plasma membrane (although a significant amount was also found in the endoplasmic reticulum). In contrast to mammalian cells, Mdr1 was not glycosylated in yeast. Nevertheless, some, if not all, of the Mdr1 made in yeast was properly folded and functional because it could be photoaffinity labeled specifically with 8-azido-ATP and because cells overexpressing Mdr1 displayed increased resistance towards valinomycin, an ionophore known to interact with Mdr1 in animal cells. Hence, a human polytopic membrane protein was correctly inserted into the yeast plasma membrane, and glycosylation was not required for its function.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	6
Volume Number	89
Language	English
Publisher	National Academy of Sciences
Publisher Date	1992-04-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Drug Resistance Genetics Membrane Glycoproteins Saccharomyces Cerevisiae Amino Acid Sequence Cell Membrane Metabolism Cloning, Molecular DNA, Recombinant Gene Expression Isolation & Purification Molecular Sequence Data P-Glycoprotein Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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