Imaging of Streptomyces coelicolor A3(2) with reduced autofluorescence reveals a novel stage of FtsZ localization.
Imaging of low abundance proteins in time and space by fluorescence microscopy is typically hampered by host-cell autofluorescence. Streptomycetes are an important model system for the study of bacterial development, and undergo multiple synchronous cell division during the sporulation stage. To analyse this phenomenon in detail, fluorescence microscopy, and in particular also the recently published novel live imaging techniques, require optimal signal to noise ratios. Here we describe the development of a novel derivative of Streptomyces coelicolor A3(2) with strongly reduced autofluorescence, allowing the imaging of fluorescently labelled proteins at significantly higher resolution. The enhanced image detail provided novel localization information for the cell division protein FtsZ, demonstrating a new developmental stage where multiple FtsZ foci accumulate at the septal plane. This suggests that multiple foci are sequentially produced, ultimately connecting to form the complete Z ring. The enhanced imaging properties are an important step forward for the confocal and live imaging of less abundant proteins and for the use of lower intensity fluorophores in streptomycetes.
Willemse
Joost
J
Department of Molecular Genetics, Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands.
van Wezel
Gilles P
GP
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000378:metabolism / Q000502:physiology*; D005456:Fluorescent Dyes / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D010641:Phenotype; D013171:Spores, Bacterial / Q000235:genetics; D048372:Streptomyces coelicolor / Q000378:metabolism*
D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; D005456:Fluorescent Dyes; C071525:FtsZ protein, Bacteria; D049452:Green Fluorescent Proteins