Watching DNA Replication Inhibitors in Action: Exploiting Time-Lapse Microfluidic Microscopy as a Tool for Target-Drug Interaction Studies in Mycobacterium

Antimicrobial Agents and Chemotherapy
Damian TrojanowskiJolanta Zakrzewska-Czerwińska

Abstract

Spreading resistance to antibiotics and the emergence of multidrug-resistant strains have become frequent in many bacterial species, including mycobacteria, which are the causative agents of severe diseases and which have profound impacts on global health. Here, we used a system of microfluidics, fluorescence microscopy, and target-tagged fluorescent reporter strains of Mycobacterium smegmatis to perform real-time monitoring of replisome and chromosome dynamics following the addition of replication-altering drugs (novobiocin, nalidixic acid, and griselimycin) at the single-cell level. We found that novobiocin stalled replication forks and caused relaxation of the nucleoid and that nalidixic acid triggered rapid replisome collapse and compaction of the nucleoid, while griselimycin caused replisome instability, with the subsequent overinitiation of chromosome replication and overrelaxation of the nucleoid. In addition to study target-drug interactions, our system also enabled us to observe how the tested antibiotics affected the physiology of mycobacterial cells (i.e., growth, chromosome segregation, etc.).

References

Nov 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A SuginoN R Cozzarelli
Oct 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·A SuginoN R Cozzarelli
Jun 1, 1989·Journal of Medical Microbiology·C S LewinJ T Smith
Oct 3, 2001·The Journal of Antimicrobial Chemotherapy·M ChatterjiV Nagaraja
Jul 1, 1956·A.M.A. Archives of Internal Medicine·W M KIRBYW D NOYES
Nov 25, 2003·The EMBO Journal·Francisco J López de SaroMike O'Donnell
Mar 30, 2004·Antimicrobial Agents and Chemotherapy·Alexandra AubryEmmanuelle Cambau
Feb 11, 2005·Chemical Reviews·Dan KahneChristopher Walsh
Jul 1, 2005·Journal of Cellular Biochemistry·Martin ThanbichlerLucy Shapiro
Jun 29, 2006·Molecular Microbiology·Muhammad MalikKarl Drlica
Nov 1, 2006·Antimicrobial Agents and Chemotherapy·Michelle M ButlerTerry L Bowlin
Nov 3, 2009·Current Genomics·Francisco J López de Saro
Jan 22, 2010·PLoS Genetics·Anjana SrivatsanJue D Wang
May 5, 2010·Nature Reviews. Microbiology·Michael A KohanskiJames J Collins
Feb 2, 2011·Proceedings of the National Academy of Sciences of the United States of America·Mohan C JoshiDavid Bates
Apr 9, 2011·Nature·Matthew A Cooper, David Shlaes
Mar 31, 2012·Expert Opinion on Drug Discovery·Gautam Sanyal, Peter Doig
Sep 13, 2012·PloS One·Graham JoyceBrian D Robertson
Feb 8, 2013·Molecular Microbiology·Bhupender SinghSantanu Dasgupta
Sep 17, 2013·Nature Communications·Isabella SantiJohn D McKinney
Oct 2, 2013·International Journal of Medical Microbiology : IJMM·Lutz Heide
Feb 1, 2014·Archives of Microbiology·Srinivasan VijayParthasarathi Ajitkumar
Mar 1, 2014·Biochemistry·Katie J AldredNeil Osheroff
Mar 7, 2014·The Journal of Biological Chemistry·Qian ChaiSuparna Sanyal
Mar 13, 2014·Microbiology·Ingvild Odsbu, Kirsten Skarstad
Jun 21, 2014·The Open Microbiology Journal·Srinivasan VijayParthasarathi Ajitkumar
Nov 20, 2014·Mediterranean Journal of Hematology and Infectious Diseases·Giorgia SulisMario C Raviglione
Dec 19, 2014·Nature Communications·M Charl MoolmanNynke H Dekker
Feb 19, 2015·MBio·Damian TrojanowskiJolanta Zakrzewska-Czerwińska
Jun 2, 2015·New Microbes and New Infections·I RocaJ Vila
Aug 6, 2015·South African Medical Journal = Suid-Afrikaanse Tydskrif Vir Geneeskunde·Marc Mendelson
Nov 14, 2015·Annual Review of Cell and Developmental Biology·Anjana BadrinarayananMichael T Laub
Jan 23, 2016·Proceedings of the National Academy of Sciences of the United States of America·Katie J AldredNeil Osheroff
May 27, 2016·Microbiology Spectrum·Jose M Munita, Cesar A Arias
Jun 18, 2016·Experimental and Therapeutic Medicine·Chenliang ZhouJing Qu

❮ Previous
Next ❯

Citations

Jul 1, 2020·Antimicrobial Agents and Chemotherapy·Nikola OjkicBartlomiej Waclaw
May 7, 2020·Frontiers in Microbiology·Joanna Hołówka, Jolanta Zakrzewska-Czerwińska
Feb 5, 2021·Scientific Reports·Marta KołodziejJolanta Zakrzewska-Czerwińska
Oct 5, 2021·Journal of Enzyme Inhibition and Medicinal Chemistry·Rania Helmy Abd El-HameedSamar Said Fatahala

❮ Previous
Next ❯

Methods Mentioned

BETA
Fluorescence
PCR

Software Mentioned

R package
CellASIC
R
ImageJ Fiji

Related Concepts

Related Feeds

CRISPR Screens in Drug Resistance

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on the application of CRISPR-Cas system in high-throughput genome-wide screens to identify genes that may confer drug resistance.