Microarray Analysis to Monitor Bacterial Cell Wall Homeostasis.
Transcriptomics, the genome-wide analysis of gene transcription, has become an important tool for characterizing and understanding the signal transduction networks operating in bacteria. Here we describe a protocol for quantifying and interpreting changes in the transcriptome of Streptomyces coelicolor that take place in response to treatment with three antibiotics active against different stages of peptidoglycan biosynthesis. The results defined the transcriptional responses associated with cell envelope homeostasis including a generalized response to all three antibiotics involving activation of transcription of the cell envelope stress sigma factor σ(E), together with elements of the stringent response, and of the heat, osmotic, and oxidative stress regulons. Many antibiotic-specific transcriptional changes were identified, representing cellular processes potentially important for tolerance to each antibiotic. The principles behind the protocol are transferable to the study of cell envelope homeostatic mechanisms probed using alternative chemical/environmental insults or in other bacterial strains.
Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University, Oxford, UK.|Department of Biochemistry, University of Cambridge, The Hopkins Building, Tennis Court Road, Cambridge, CB2 1QW, UK.
Hee-Jeon
HJ
Hong
Department of Biochemistry, Cambridge Systems Biology Centre, University of Cambridge, The Sanger Building, 80 Tennis Court Road, Cambridge, CB2 1GA, UK. arh69@cam.ac.uk.|Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK. arh69@cam.ac.uk.
Andy
A
Hesketh
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics*; D002473:Cell Wall / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D006706:Homeostasis; D006358:Hot Temperature; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D018384:Oxidative Stress; D010457:Peptidoglycan / Q000096:biosynthesis; D048372:Streptomyces coelicolor / Q000187:drug effects / Q000235:genetics*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D010457:Peptidoglycan
Antibiotic; Cell envelope homeostasis; Streptomyces; Transcriptomics