Determination of mRNA half-lives in Candida albicans using thiolutin as a transcription inhibitor
- PMID: 17036064
- DOI: 10.1139/g06-046
Determination of mRNA half-lives in Candida albicans using thiolutin as a transcription inhibitor
Abstract
A method for determining mRNA half-lives in the polymorphic fungus Candida albicans is described. It employs growth in a defined medium, the inhibition of transcription with thiolutin (10-20 microg/mL), and quantitative Northern blotting. The method is effective for the A72, SC5314, and CAI-4 strains of C. albicans, and for mRNAs that have a wide variety of decay rates and steady-state abundances. The range of half-lives detected (from 4-168 min) shows that this method is effective for mRNAs with widely varying half-lives. The mRNA decay rates obtained are compared with those for orthologous mRNAs from Saccharomyces cerevisiae. This procedure should work for a broad range of C. albicans strains and can be adapted to other fungal species.
Similar articles
-
The transcriptional inhibitor thiolutin blocks mRNA degradation in yeast.Yeast. 2008 Feb;25(2):85-92. doi: 10.1002/yea.1548. Yeast. 2008. PMID: 17914747
-
Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae.Methods Mol Biol. 2017;1507:169-177. doi: 10.1007/978-1-4939-6518-2_13. Methods Mol Biol. 2017. PMID: 27832540
-
Measurement of mRNA decay rates in Saccharomyces cerevisiae using rpb1-1 strains.J Vis Exp. 2014 Dec 13;(94):52240. doi: 10.3791/52240. J Vis Exp. 2014. PMID: 25549102 Free PMC article.
-
mRNA abundance and half-life measurements.Methods Mol Biol. 1996;53:277-95. doi: 10.1385/0-89603-319-8:277. Methods Mol Biol. 1996. PMID: 8924989 Review. No abstract available.
-
Messenger RNA stability in yeast.Yeast. 1989 Jul-Aug;5(4):239-57. doi: 10.1002/yea.320050405. Yeast. 1989. PMID: 2551103 Review. No abstract available.
Cited by
-
The Association of Rpb4 with RNA Polymerase II Depends on CTD Ser5P Phosphatase Rtr1 and Influences mRNA Decay in Saccharomyces cerevisiae.Int J Mol Sci. 2022 Feb 11;23(4):2002. doi: 10.3390/ijms23042002. Int J Mol Sci. 2022. PMID: 35216121 Free PMC article.
-
Blockade of deubiquitinating enzyme PSMD14 overcomes chemoresistance in head and neck squamous cell carcinoma by antagonizing E2F1/Akt/SOX2-mediated stemness.Theranostics. 2021 Jan 1;11(6):2655-2669. doi: 10.7150/thno.48375. eCollection 2021. Theranostics. 2021. PMID: 33456565 Free PMC article.
-
Fungal community analysis by high-throughput sequencing of amplified markers--a user's guide.New Phytol. 2013 Jul;199(1):288-299. doi: 10.1111/nph.12243. Epub 2013 Mar 28. New Phytol. 2013. PMID: 23534863 Free PMC article. Review.
-
Multiplexed target enrichment of coding and non-coding transcriptomes enables studying Candida spp. infections from human derived samples.Front Cell Infect Microbiol. 2023 Jan 24;13:1093178. doi: 10.3389/fcimb.2023.1093178. eCollection 2023. Front Cell Infect Microbiol. 2023. PMID: 36761895 Free PMC article.
-
Dithiolopyrrolones: biosynthesis, synthesis, and activity of a unique class of disulfide-containing antibiotics.Nat Prod Rep. 2014 Jul;31(7):905-23. doi: 10.1039/c3np70106a. Nat Prod Rep. 2014. PMID: 24835149 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
