title: The conserved ATPase Get3/Arr4 modulates the activity of membrane-associated proteins in Saccharomyces cerevisiae.
abstract: The regulation of cellular membrane dynamics is crucial for maintaining proper cell growth and division. The Cdc48-Npl4-Ufd1 complex is required for several regulated membrane-associated processes as part of the ubiquitin-proteasome system, including ER-associated degradation and the control of lipid composition in yeast. In this study we report the results of a genetic screen in Saccharomyces cerevisiae for extragenic suppressors of a temperature-sensitive npl4 allele and the subsequent analysis of one suppressor, GET3/ARR4. The GET3 gene encodes an ATPase with homology to the regulatory component of the bacterial arsenic pump. Mutants of GET3 rescue several phenotypes of the npl4 mutant and transcription of GET3 is coregulated with the proteasome, illustrating a functional relationship between GET3 and NPL4 in the ubiquitin-proteasome system. We have further found that Get3 biochemically interacts with the trans-membrane domain proteins Get1/Mdm39 and Get2/Rmd7 and that Deltaget3 is able to suppress phenotypes of get1 and get2 mutants, including sporulation defects. In combination, our characterization of GET3 genetic and biochemical interactions with NPL4, GET1, and GET2 implicates Get3 in multiple membrane-dependent pathways.
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Auld
authors.forename: Amy L
authors.initials: AL
authors.lastname: Hitchcock
authors.forename: Hugh K
authors.initials: HK
authors.lastname: Doherty
authors.forename: Seth
authors.initials: S
authors.lastname: Frietze
authors.forename: Linda S
authors.initials: LS
authors.lastname: Huang
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Silver
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D046912:Multiprotein Complexes / Q000378:metabolism; D009154:Mutation; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D029543:Nucleocytoplasmic Transport Proteins; D010641:Phenotype; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D013172:Spores, Fungal / Q000235:genetics; D013489:Suppression, Genetic / Q000502:physiology; D014018:Tissue Distribution; D000074405:Valosin Containing Protein; D033921:Vesicular Transport Proteins
chemical_list: C113359:ARR4 protein, Arabidopsis; D033942:Adaptor Proteins, Vesicular Transport; D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; C503510:GET1 protein, S cerevisiae; C515526:Get2 protein, S cerevisiae; D008565:Membrane Proteins; D046912:Multiprotein Complexes; C104361:NPL4 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D029543:Nucleocytoplasmic Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; C444652:UFD1 protein, S cerevisiae; D033921:Vesicular Transport Proteins; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases; C000614177:CDC48 protein, S cerevisiae; D000074405:Valosin Containing Protein