Yeast transformation mediated by Agrobacterium strains harboring an Ri plasmid: comparative study between  GALLS  of an Ri plasmid and virE  of a Ti plasmid.
Agrobacterium strains containing a Ti plasmid can transfer T-DNA not only to plants but also to fungi, including the yeast Saccharomyces cerevisiae. However, no Agrobacterium strain harboring an Ri plasmid has been evaluated in fungal transformation. Some Ri plasmids have GALLS , instead of virE1 and virE2. GALLS protein can functionally substitute in plant transformation for a structurally different protein VirE2. In this study, we compared the yeast transformation ability among Agrobacterium donors: a strain containing a Ti plasmid, strains harboring either an agropine-type or a mikimopine-type Ri plasmid, and a strain having a modified Ri plasmid supplemented with a Ti plasmid type virE operon. Agrobacterium strains possessing GALLS transformed yeast cells far less efficiently than the strain containing virE operon. Production of GALLS in recipient yeast cells improved the yeast transformation mediated by an Agrobacterium strain lacking neither GALLS nor virE operon. A reporter assay to detect mobilization of the proteins fused with Cre recombinase revealed that VirE2 protein is much more abundant in yeast cells than GALLS. Based on these results, we concluded that the low yeast transformability mediated by Agrobacterium strains having the Ri plasmid is because of low amount of mobilized GALLS in yeast cells.
Department of Biological Science, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Hiroshima, 739-8526, Japan.
Kazuya
K
Kiyokawa
Shinji
S
Yamamoto
Yukari
Y
Sato
Naoto
N
Momota
Katsuyuki
K
Tanaka
Kazuki
K
Moriguchi
Katsunori
K
Suzuki
D060054:Agrobacterium / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015966:Gene Expression Regulation, Fungal; D023061:Gene Order; D009876:Operon / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D014170:Transformation, Genetic
D001426:Bacterial Proteins