Overexpression of Bud5p can suppress mutations in the Gsp1p guanine nucleotide exchange factor Prp20p in Saccharomyces cerevisiae.
The gene product Prp20p, which is located in the nucleus, serves as the nucleotide exchange factor (GEF) for the small nuclear G protein Gsp1p in Saccharomyces cerevisiae, and catalyses the replacement of Gsp1-bound GDP by GTP. These proteins are involved in numerous cellular processes, including nucleocytoplasmic trafficking of macromolecules, cell cycle progression, DNA replication and maintenance of chromosome structure/stability. It is believed that in order to complete a full GDP/GTP cycle, Gsp1p has to shuttle between the nucleus and the cytoplasm, where its GTPase Activating Protein (GAP) Rna1p is located. Here, we report on the ability of Bud5p, the exchange factor for Rsr1p, to suppress conditional prp20 mutants when an extra copy of GSP1 is present. This suppression by BUD5 can be reversed by simultaneous overexpression of RNA1, and is not Rsr1p-dependent, nor allele-specific. We also show that Bud5p can physically interact with Gsplp, both in vitro and in vivo. These,findings raise the possibility that Bud5p could act as a cytoplasmic exchange factor for Gsp1p and, therefore, that a complete GDP/GTP cycle could take place in the cytoplasm.
Clément
M
M
Département de Microbiologie et Immunologie, Université de Montréal, Québec, Canada.
Lavallée
F
F
Barbès-Morin
G
G
de Repentigny
L
L
Belhumeur
P
P
D001483:Base Sequence; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D015870:Gene Expression; D005800:Genes, Fungal; D016153:Genes, Suppressor; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics*; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics*; D011233:Precipitin Tests; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins
C091391:BUD5 protein, S cerevisiae; D017931:DNA Primers; D004268:DNA-Binding Proteins; C490024:FUS1 protein, S cerevisiae; D005656:Fungal Proteins; C080324:GSP1 protein, S cerevisiae; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; D009687:Nuclear Proteins; C067050:SRM1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D020559:Monomeric GTP-Binding Proteins