Open AccessReconstitution of a bacterial/plant polyamine biosynthesis pathway in Saccharomyces cerevisiae
Author(s) -
Ronald D. Klein,
Timothy G. Geary,
A. S. Gibson,
Margaret Anne Favreau,
C.A. Winterrowd,
Steve J. Upton,
Jan S. Keithly,
Guangxi Zhu,
Russell L. Malmberg,
Martha P. Martinez,
Nigel Yarlett
Publication year - 1999
Publication title -
microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.019
H-Index - 179
eISSN - 1465-2080
pISSN - 1350-0872
DOI - 10.1099/13500872-145-2-301
Subject(s) - arginine decarboxylase , biochemistry , agmatine , ornithine decarboxylase , saccharomyces cerevisiae , polyamine , putrescine , biology , escherichia coli , cadaverine , complementation , enzyme , ornithine decarboxylase antizyme , biosynthesis , yeast , plasmid , gene , mutant
Polyamine synthesis in most organisms is initiated by the decarboxylation of ornithine to form putrescine via ornithine decarboxylase (ODC). Plants, some bacteria and some fungi and protozoa generate putrescine from arginine, via arginine decarboxylase (ADC) and agmatine ureohydrolase (AUH) or agmatine iminohydrolase. A polyamine-requiring strain of Saccharomyces cerevisiae with a mutation in the gene encoding ODC was transformed with plasmids bearing genes encoding Escherichia coli ADC and AUH. Transformants regained the ability to grow in the absence of exogenous polyamines and contained enzyme activities consistent with the presence of both prokaryotic enzymes. Similar results were obtained when a plasmid containing a gene encoding oat (Avena sativa L.) ADC was substituted for the E. coli gene. These data demonstrate the successful complementation of a yeast biosynthetic polyamine synthesis defect by genes encoding an alternative pathway found in bacteria; they also show that plant ADC can substitute for the bacterial enzyme in this pathway. The recombinant yeast provides a tool for the study of the functional properties of these enzymes and for discovery of compounds that specifically inhibit this pathway.