Gene Annotation and Drug Target Discovery in Candida albicans with a Tagged Transposon Mutant Collection

Oh, Julia; Fung, Eula; Schlecht, Ulrich; Davis, Ronald W.; Giaever, Guri; St. Onge, Robert P.; Deutschbauer, Adam; Nislow, Corey

doi:10.1371/journal.ppat.1001140

Title: **Gene Annotation and Drug Target Discovery in Candida albicans with a Tagged Transposon Mutant Collection**

Journal Article · Thu Oct 07 00:00:00 EDT 2010 · PLoS Pathogens

DOI:https://doi.org/10.1371/journal.ppat.1001140· OSTI ID:1627897

Oh, Julia ^[1]; Fung, Eula ^[2]; Schlecht, Ulrich ^[2]; Davis, Ronald W. ^[3]; Giaever, Guri ^[4]; St. Onge, Robert P. ^[2]; Deutschbauer, Adam ^[5]; Nislow, Corey ^[6]

Stanford University, CA (United States)
Stanford Genome Technology Center, Palo Alto, CA (United States)
Stanford University, CA (United States); Stanford Genome Technology Center, Palo Alto, CA (United States)
University of Toronto, ON (Canada); Donnelley Center for Cellular and Biomolecular Research, Toronto, ON (Canada)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Univ. of Toronto, ON (Canada); Donnelley Center for Cellular and Biomolecular Research, Toronto, ON (Canada)

Candida albicans is the most common human fungal pathogen, causing infections that can be lethal in immunocompromised patients. Although Saccharomyces cerevisiae has been used as a model for C. albicans, it lacks C. albicans’ diverse morphogenic forms and is primarily non-pathogenic. Comprehensive genetic analyses that have been instrumental for determining gene function in S. cerevisiae are hampered in C. albicans, due in part to limited resources to systematically assay phenotypes of loss-of-function alleles. Here, we constructed and screened a library of 3633 tagged heterozygous transposon disruption mutants, using them in a competitive growth assay to examine nutrient- and drug-dependent haploinsufficiency. We identified 269 genes that were haploinsufficient in four growth conditions, the majority of which were condition-specific. These screens identified two new genes necessary for filamentous growth as well as ten genes that function in essential processes. We also screened 57 chemically diverse compounds that more potently inhibited growth of C. albicans versus S. cerevisiae. For four of these compounds, we examined the genetic basis of this differential inhibition. Notably, Sec7p was identified as the target of brefeldin A in C. albicans screens, while S. cerevisiae screens with this compound failed to identify this target. We also uncovered a new C. albicans-specific target, Tfp1p, for the synthetic compound 0136-0228. These results highlight the value of haploinsufficiency screens directly in this pathogen for gene annotation and drug target identification.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Human Genome Research Institute (NHGRI); Canadian Cancer Society; Stanford Genome Training Program; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-05CH11231; HG000205; RO1 HG003317; CIHR MOP-81340; CIHR MOP-84305; 020380; T32 HG00044; P01 GH000205

OSTI ID:: 1627897

Journal Information:: PLoS Pathogens, Vol. 6, Issue 10; ISSN 1553-7374

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Candida albicans
saccharomyces cerevisiae
gene pool
haploinsufficiency
genomics
transposable elements
drug screening
heterozygosity

Title: Gene Annotation and Drug Target Discovery in Candida albicans with a Tagged Transposon Mutant Collection

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Title: **Gene Annotation and Drug Target Discovery in Candida albicans with a Tagged Transposon Mutant Collection**