Functional analysis of the Saccharomyces cerevisiae DUP240 multigene family reveals membrane-associated proteins that are not essential for cell viability

• Published in: Microbiology (Society for General Microbiology) Vol. 148; no. 7; pp. 2111 - 2123
• Main Authors: Poirey, Remy, Despons, Laurence, Leh, Veronique, Lafuente, Maria-Jose, Potier, Serge, Souciet, Jean-Luc, Jauniaux, Jean-Claude
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.07.2002; Society for General Microbiology
• Subjects: Amino Acid Sequence; Biological and medical sciences; Cell Membrane - metabolism; DUP240 gene; Fundamental and applied biological sciences. Psychology; Gene Deletion; Genes, Essential; Green Fluorescent Proteins; Growth, nutrition, metabolism, transports, enzymes. Molecular biology; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Membrane Proteins - genetics; Membrane Proteins - metabolism; Microbiology; Multigene Family; Mycology; Recombinant Fusion Proteins; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Sequence Analysis, DNA; Sos1 protein; Subcellular Fractions - metabolism; Tandem Repeat Sequences - genetics; Transformation, Genetic; Two-Hybrid System Techniques; Membrane protein; Yeast; Genetic marker; Site directed mutagenesis; Fluorescence; Saccharomyces bayanus; Fungi; Characterization; Deletion; Multigene family; Ascomycetes; Kinetics; Concentrate; Saccharomyces cerevisiae; Thallophyta
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/00221287-148-7-2111

Angewandte Tumorvirologie, Abteilung F0100 and Virologie Appliquée à l’Oncologie (Unité INSERM 375), Deutsches Krebsforschungszentrum, P. 1011949, D-69009 Heidelberg, Germany 1 Laboratoire de Génétique et Microbiologie, UPRES-A 7010 ULP/CNRS, Institut de Botanique, 28 rue Goethe, F-67083 Strasbourg cedex, France 2 Author for correspondence: Jean-Claude Jauniaux. Tel: +49 6221 42 49 71. Fax: +49 6221 42 52 49 71. e-mail: j.jauniaux{at}dkfz.de The DUP240 gene family of Saccharomyces cerevisiae is composed of 10 members. They encode proteins of about 240 amino acids which contain two predicted transmembrane domains. Database searches identified only one homologue in the closely related species Saccharomyces bayanus , indicating that the DUP240 genes encode proteins specific to Saccharomyces sensu stricto . The short-flanking homology PCR gene-replacement strategy with a variety of selective markers for replacements, and classical genetic methods, were used to generate strains deleted for all 10 DUP240 genes. All of the knock-out strains were viable and had similar growth kinetics to the wild-type. Two-hybrid screens, hSos1p fusions and GFP fusions were carried out; the results indicated that the Dup240 proteins are membrane associated, and that some of them are concentrated around the plasma membrane. Keywords: gene disruption, membrane protein, gene tandem repeats, two-hybrid system, yeast Abbreviations: GFP, green fluorescent protein a Present address: Centro de Biologia Molecular Severo Ochoa, Universidad Autonoma, Canto Blanco, 28049, Madrid, Spain.