Purine synthesis and increased Agrobacterium tumefaciens transformation of yeast and plants.
The bacterium Agrobacterium tumefaciens transforms eukaryotic hosts by transferring DNA to the recipient cell where it is integrated and expressed. Bacterial factors involved in this interkingdom gene transfer have been described, but less is known about host-cell factors. Using the yeast Saccharomyces cerevisiae as a model host, we devised a genetic screen to identify yeast mutants with altered transformation sensitivities. Twenty-four adenine auxotrophs were identified that exhibited supersensitivity to A. tumefaciens-mediated transformation when deprived of adenine. We extended these results to plants by showing that purine synthesis inhibitors cause supersensitivity to A. tumefaciens transformation in three plant species. The magnitude of this effect is large and does not depend on prior genetic manipulations of host cells. These data indicate the utility of yeast as a model for the transformation process and identify purine biosynthesis as a key determinant of transformation efficiency. These findings should increase the utility of A. tumefaciens in genetic engineering.
Roberts
Radclyffe L
RL
Department of Microbiology, University of Washington, Seattle 98195-7242, USA.
Metz
Matthew
M
Monks
Dave E
DE
Mullaney
Matthew Lockwood
ML
Hall
Tyler
T
Nester
Eugene W
EW
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000472:pathogenicity; D018744:DNA, Plant / Q000235:genetics; D011687:Purines / Q000096:biosynthesis*; D014170:Transformation, Genetic; D014774:Virulence / Q000235:genetics
D018744:DNA, Plant; D011687:Purines