Characterizing Candida glabrata Pdr1, a Hyperactive Transcription Factor Involved in Azole Resistance.
Animals
Humans
Transcription Factors
/ genetics
Azoles
/ pharmacology
Candida glabrata
/ genetics
DNA-Binding Proteins
/ metabolism
Fungal Proteins
/ genetics
Gene Expression Regulation, Fungal
Drug Resistance, Fungal
/ genetics
Antifungal Agents
/ pharmacology
Fluconazole
/ pharmacology
Microbial Sensitivity Tests
Azole resistance
Candida glabrata
Isogenic strains
Marker-less integration of PDR1-GOF into the genome
Transcription factor Pdr1
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2023
2023
Historique:
medline:
10
4
2023
entrez:
6
4
2023
pubmed:
7
4
2023
Statut:
ppublish
Résumé
This chapter illustrates how to prepare isogenic strains carrying gain-of-function forms of transcription factor Pdr1 in the human pathogen Candida glabrata. Simple steps are described that lead from a characterized plasmid-borne PDR1-GOF allele to its integration into the yeast genome in a markerless manner. Pdr1-GOF strains constructed by this approach are suitable for virulence studies in an animal host.
Identifiants
pubmed: 37024701
doi: 10.1007/978-1-0716-3155-3_11
doi:
Substances chimiques
Transcription Factors
0
Azoles
0
DNA-Binding Proteins
0
Fungal Proteins
0
Antifungal Agents
0
Fluconazole
8VZV102JFY
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
169-179Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Tsai HF, Krol AA, Sarti KE, Bennett JE (2006) Candida glabrata PDR1, a transcriptional regulator of a pleiotropic drug resistance network, mediates azole resistance in clinical isolates and petite mutants. Antimicrob Agents Chemother 50(4):1384–1392. https://doi.org/10.1128/AAC.50.4.1384-1392.2006
doi: 10.1128/AAC.50.4.1384-1392.2006
pubmed: 16569856
pmcid: 1426987
Vermitsky JP, Edlind TD (2004) Azole resistance in Candida glabrata: coordinate upregulation of multidrug transporters and evidence for a Pdr1-like transcription factor. Antimicrob Agents Chemother 48(10):3773–3781. https://doi.org/10.1128/AAC.48.10.3773-3781.2004
doi: 10.1128/AAC.48.10.3773-3781.2004
pubmed: 15388433
pmcid: 521908
Whaley SG, Berkow EL, Rybak JM, Nishimoto AT, Barker KS, Rogers PD (2016) Azole antifungal resistance in Candida albicans and emerging non-albicans Candida species. Front Microbiol 7:2173. https://doi.org/10.3389/fmicb.2016.02173
doi: 10.3389/fmicb.2016.02173
pubmed: 28127295
Bennett JE, Izumikawa K, Marr KA (2004) Mechanism of increased fluconazole resistance in Candida glabrata during prophylaxis. Antimicrob Agents Chemother 48(5):1773–1777. https://doi.org/10.1128/aac.48.5.1773-1777.2004
doi: 10.1128/aac.48.5.1773-1777.2004
pubmed: 15105134
pmcid: 400565
Ferrari S, Ischer F, Calabrese D, Posteraro B, Sanguinetti M, Fadda G, Rohde B, Bauser C, Bader O, Sanglard D (2009) Gain of function mutations in CgPDR1 of Candida glabrata not only mediate antifungal resistance but also enhance virulence. PLoS Pathog 5(1):e1000268. https://doi.org/10.1371/journal.ppat.1000268
doi: 10.1371/journal.ppat.1000268
pubmed: 19148266
pmcid: 2607542
Tsai HF, Sammons LR, Zhang X, Suffis SD, Su Q, Myers TG, Marr KA, Bennett JE (2010) Microarray and molecular analyses of the azole resistance mechanism in Candida glabrata oropharyngeal isolates. Antimicrob Agents Chemother 54(8):3308–3317. https://doi.org/10.1128/AAC.00535-10
doi: 10.1128/AAC.00535-10
pubmed: 20547810
pmcid: 2916311
Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las PA, Iluore J, Cormack BP (2013) Expression plasmids for use in Candida glabrata. G3 (Bethesda) 3(10):1675–1686. https://doi.org/10.1534/g3.113.006908
doi: 10.1534/g3.113.006908
pubmed: 23934995
Zheng L, Baumann U, Reymond JL (2004) An efficient one-step site-directed and site-saturation mutagenesis protocol. Nucleic Acids Res 32(14):e115. https://doi.org/10.1093/nar/gnh110
doi: 10.1093/nar/gnh110
pubmed: 15304544
pmcid: 514394
Khakhina S, Simonicova L, Moye-Rowley WS (2018) Positive autoregulation and repression of transactivation are key regulatory features of the Candida glabrata Pdr1 transcription factor. Mol Microbiol 107(6):747–764. https://doi.org/10.1111/mmi.13913
doi: 10.1111/mmi.13913
pubmed: 29363861
pmcid: 5842128
Simonicova L, Moye-Rowley WS (2020) Functional information from clinically-derived drug resistant forms of the Candida glabrata Pdr1 transcription factor. PLoS Genet 16(8):e1009005. https://doi.org/10.1371/journal.pgen.1009005
doi: 10.1371/journal.pgen.1009005
pubmed: 32841236
pmcid: 7473514
Ferrari S, Sanguinetti M, Torelli R, Posteraro B, Sanglard D (2011) Contribution of CgPDR1-regulated genes in enhanced virulence of azole-resistant Candida glabrata. PLoS One 6(3):e17589. https://doi.org/10.1371/journal.pone.0017589
doi: 10.1371/journal.pone.0017589
pubmed: 21408004
pmcid: 3052359
Vu BG, Moye-Rowley WS (2018) Construction and use of a recyclable marker to examine the role of major facilitator superfamily protein members in candida glabrata drug resistance phenotypes. mSphere 3(2):e00099. https://doi.org/10.1128/mSphere.00099-18
doi: 10.1128/mSphere.00099-18
pubmed: 29600281
pmcid: 5874441
Gietz RD, Schiestl RH (2007) High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2(1):31–34. https://doi.org/10.1038/nprot.2007.13
doi: 10.1038/nprot.2007.13
pubmed: 17401334
Paul S, Bair TB, Moye-Rowley WS (2014) Identification of genomic binding sites for Candida glabrata Pdr1 transcription factor in wild-type and rho0 cells. Antimicrob Agents Chemother 58(11):6904–6912. https://doi.org/10.1128/AAC.03921-14
doi: 10.1128/AAC.03921-14
pubmed: 25199772
pmcid: 4249425
Vu BG, Thomas GH, Moye-Rowley WS (2019) Evidence that Ergosterol biosynthesis modulates activity of the Pdr1 transcription factor in Candida glabrata. mBio 10:3. https://doi.org/10.1128/mBio.00934-19
doi: 10.1128/mBio.00934-19