Fluorescence-based reporter for gauging cyclic di-GMP levels in Pseudomonas aeruginosa.

Applied and Environmental Microbiology
Morten T RybtkeTim Tolker-Nielsen

Abstract

The increased tolerance toward the host immune system and antibiotics displayed by biofilm-forming Pseudomonas aeruginosa and other bacteria in chronic infections such as cystic fibrosis bronchopneumonia is of major concern. Targeting of biofilm formation is believed to be a key aspect in the development of novel antipathogenic drugs that can augment the effect of classic antibiotics by decreasing antimicrobial tolerance. The second messenger cyclic di-GMP is a positive regulator of biofilm formation, and cyclic di-GMP signaling is now regarded as a potential target for the development of antipathogenic compounds. Here we describe the development of fluorescent monitors that can gauge the cellular level of cyclic di-GMP in P. aeruginosa. We have created cyclic di-GMP level reporters by transcriptionally fusing the cyclic di-GMP-responsive cdrA promoter to genes encoding green fluorescent protein. We show that the reporter constructs give a fluorescent readout of the intracellular level of cyclic di-GMP in P. aeruginosa strains with different levels of cyclic di-GMP. Furthermore, we show that the reporters are able to detect increased turnover of cyclic di-GMP mediated by treatment of P. aeruginosa with the phosphodiesterase ind...Continue Reading

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Citations

Feb 14, 2013·Antimicrobial Agents and Chemotherapy·Song Lin ChuaMichael Givskov
Mar 9, 2013·Microbiology and Molecular Biology Reviews : MMBR·Ute RömlingMark Gomelsky
Oct 27, 2015·Journal of Applied Microbiology·S Vishnu PawarLivia Leoni
Mar 7, 2014·Environmental Microbiology·Mustafa FazliTim Tolker-Nielsen
Apr 15, 2015·Environmental Microbiology Reports·Alicia Jiménez-FernándezFernando Govantes
May 28, 2014·FEMS Microbiology Letters·Emeline BouffartiguesSylvie Chevalier
May 18, 2016·FEMS Microbiology Letters·Aroa López-SánchezFernando Govantes
Aug 5, 2016·Frontiers in Microbiology·María Isabel Ramos-GonzálezManuel Espinosa-Urgel
Aug 2, 2016·Environmental Science & Technology·Sanly LiuRose Amal
Sep 28, 2016·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Angelika Gründling, Vincent T Lee
Dec 15, 2015·Environmental Microbiology·Andrés Corral-LugoTino Krell
Apr 28, 2016·Environmental Microbiology Reports·Yujie XiaoQiaoyun Huang
Feb 7, 2017·Nature Reviews. Microbiology·Urs JenalChristian Lori
Feb 11, 2018·Applied and Environmental Microbiology·Anne E MattinglyJoshua D Shrout
Sep 14, 2017·International Journal of Molecular Sciences·Tim Holm JakobsenMichael Givskov
Dec 23, 2017·Applied and Environmental Microbiology·Kasper Nørskov KraghThomas Bjarnsholt
Oct 17, 2017·Molecular Microbiology·Yun Heacock-KangTung T Hoang
Jul 12, 2018·Microbiology·Sanjida Halim TopaLinda L Blackall
Apr 22, 2015·Journal of Bacteriology·Henrik AlmbladTim Tolker-Nielsen
Feb 1, 2017·Antimicrobial Agents and Chemotherapy·Lise Goltermann, Tim Tolker-Nielsen

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