Functional analysis of the Saccharomyces cerevisiae DUP240 multigene family reveals membrane-associated proteins that are not essential for cell viability.
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.
Poirey
Rémy
R
Laboratoire de Génétique et Microbiologie, UPRES-A 7010 ULP/CNRS, Institut de Botanique, 28 rue Goethe, F-67083 Strasbourg cedex, France2.|Angewandte Tumorvirologie, Abteilung F0100 and Virologie Appliquée à l'Oncologie (Unité INSERM 375), Deutsches Krebsforschungszentrum, P. 1011949, D-69009 Heidelberg, Germany1.
Despons
Laurence
L
Laboratoire de Génétique et Microbiologie, UPRES-A 7010 ULP/CNRS, Institut de Botanique, 28 rue Goethe, F-67083 Strasbourg cedex, France2.
Leh
Véronique
V
Laboratoire de Génétique et Microbiologie, UPRES-A 7010 ULP/CNRS, Institut de Botanique, 28 rue Goethe, F-67083 Strasbourg cedex, France2.
Lafuente
Maria-Jose
MJ
Angewandte Tumorvirologie, Abteilung F0100 and Virologie Appliquée à l'Oncologie (Unité INSERM 375), Deutsches Krebsforschungszentrum, P. 1011949, D-69009 Heidelberg, Germany1.
Potier
Serge
S
Laboratoire de Génétique et Microbiologie, UPRES-A 7010 ULP/CNRS, Institut de Botanique, 28 rue Goethe, F-67083 Strasbourg cedex, France2.
Souciet
Jean-Luc
JL
Laboratoire de Génétique et Microbiologie, UPRES-A 7010 ULP/CNRS, Institut de Botanique, 28 rue Goethe, F-67083 Strasbourg cedex, France2.
Jauniaux
Jean-Claude
JC
Angewandte Tumorvirologie, Abteilung F0100 and Virologie Appliquée à l'Oncologie (Unité INSERM 375), Deutsches Krebsforschungszentrum, P. 1011949, D-69009 Heidelberg, Germany1.
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism; D017353:Gene Deletion; D020043:Genes, Essential; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013347:Subcellular Fractions / Q000378:metabolism; D020080:Tandem Repeat Sequences / Q000235:genetics; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques
D008164:Luminescent Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins