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酿酒酵母在液体培养基中生物膜形成的遗传基础。

Genetic basis for Saccharomyces cerevisiae biofilm in liquid medium.

作者信息

Scherz Kaj, Bojsen Rasmus, Gro Laura, Weiss Martin, Lisby Michael, Folkesson Anders, Regenberg Birgitte

机构信息

Department of Biology, University of Copenhagen, Copenhagen, Denmark Department of Systems Biology, Technical University of Denmark, Copenhagen, Denmark.

Department of Biology, University of Copenhagen, Copenhagen, Denmark.

出版信息

G3 (Bethesda). 2014 Jul 9;4(9):1671-80. doi: 10.1534/g3.114.010892.

Abstract

Biofilm-forming microorganisms switch between two forms: free-living planktonic and sessile multicellular. Sessile communities of yeast biofilms in liquid medium provide a primitive example of multicellularity and are clinically important because biofilms tend to have other growth characteristics than free-living cells. We investigated the genetic basis for yeast, Saccharomyces cerevisiae, biofilm on solid surfaces in liquid medium by screening a comprehensive deletion mutant collection in the Σ1278b background and found 71 genes that were essential for biofilm development. Quantitative northern blots further revealed that AIM1, ASG1, AVT1, DRN1, ELP4, FLO8, FMP10, HMT1, KAR5, MIT1, MRPL32, MSS11, NCP1, NPR1, PEP5, PEX25, RIM8, RIM101, RGT1, SNF8, SPC2, STB6, STP22, TEC1, VID24, VPS20, VTC3, YBL029W, YBL029C-A, YFL054C, YGR161W-C, YIL014C-A, YIR024C, YKL151C, YNL200C, YOR034C-A, and YOR223W controlled biofilm through FLO11 induction. Almost all deletion mutants that were unable to form biofilms in liquid medium also lost the ability to form surface-spreading biofilm colonies (mats) on agar and 69% also lost the ability to grow invasively. The protein kinase A isoform Tpk3p functioned specifically in biofilm and mat formation. In a tpk3 mutant, transcription of FLO11 was induced three-fold compared with wild-type, but biofilm development and cell-cell adhesion was absent, suggesting that Tpk3p regulates FLO11 positive posttranscriptionally and negative transcriptionally.The study provides a resource of biofilm-influencing genes for additional research on biofilm development and suggests that the regulation of FLO11 is more complex than previously anticipated.

摘要

形成生物膜的微生物在两种形式之间转换

自由生活的浮游形式和固着的多细胞形式。液体培养基中酵母生物膜的固着群落提供了多细胞性的原始例子,并且在临床上很重要,因为生物膜往往具有与自由生活细胞不同的其他生长特性。我们通过筛选Σ1278b背景下的全面缺失突变体文库,研究了液体培养基中固体表面上酿酒酵母生物膜形成的遗传基础,发现了71个对生物膜发育至关重要的基因。定量Northern印迹进一步揭示,AIM1、ASG1、AVT1、DRN1、ELP4、FLO8、FMP10、HMT1、KAR5、MIT1、MRPL32、MSS11、NCP1、NPR1、PEP5、PEX25、RIM8、RIM101、RGT1、SNF8、SPC2、STB6、STP22、TEC1、VID24、VPS20、VTC3、YBL029W、YBL029C-A、YFL054C、YGR161W-C、YIL014C-A、YIR024C、YKL151C、YNL200C、YOR034C-A和YOR223W通过诱导FLO11来控制生物膜。几乎所有在液体培养基中无法形成生物膜的缺失突变体也失去了在琼脂上形成表面扩散生物膜菌落(菌苔)的能力,69%的突变体还失去了侵袭性生长的能力。蛋白激酶A亚型Tpk3p在生物膜和菌苔形成中具有特异性功能。在tpk3突变体中,与野生型相比,FLO11的转录诱导了三倍,但生物膜发育和细胞间粘附缺失,这表明Tpk3p在转录后对FLO11起正向调节作用,在转录水平上起负向调节作用。该研究为生物膜发育的进一步研究提供了影响生物膜的基因资源,并表明FLO11的调控比以前预期的更为复杂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/617a/4169159/f11e176dfddd/1671f1.jpg

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