Yeast as a model system to study drugs effective against apicomplexan proteins.
Biochemical and genetic analyses are required to identify potential drug targets in apicomplexan parasites, but these studies have proved difficult in most parasite systems. We have developed methods based on expression of parasite proteins in the budding yeast, Saccharomyces cerevisiae, to rapidly screen drugs directed against particular parasite targets, to study the structure and function of these target molecules, and to identify mutations in the parasite genes that alter enzyme specificity or drug sensitivity. In this paper we outline the parameters that need to be considered to design yeast strains that function efficiently to assay function of parasite proteins. Basic protocols and methods are included. We detail some problems that might be encountered in the engineering of these yeast strains and suggest possible solutions.
Sibley
C H
CH
Department of Genetics, University of Washington, Seattle, Washington, 98195-7360, USA. sibley@genetics.washington.edu
Brophy
V H
VH
Cheesman
S
S
Hamilton
K L
KL
Hankins
E G
EG
Wooden
J M
JM
Kilbey
B
B
D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D016782:Apicomplexa / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis / Q000235:genetics*; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004351:Drug Resistance; D004798:Enzymes / Q000096:biosynthesis / Q000235:genetics; D017125:Genes, Protozoan; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics; D011739:Pyrimethamine / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic
D000981:Antiprotozoal Agents; D004798:Enzymes; D015800:Protozoan Proteins; D012333:RNA, Messenger; D004250:DNA Topoisomerases, Type II; D011739:Pyrimethamine