Dynamic localization of membrane proteins in Bacillus subtilis

• Published in: Microbiology (Society for General Microbiology) Vol. 150; no. 9; pp. 2815 - 2824
• Main Authors: Johnson, A. S, van Horck, S, Lewis, P. J
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.09.2004; Society for General Microbiology
• Subjects: Artificial Gene Fusion; ATP Synthetase Complexes - analysis; ATP Synthetase Complexes - genetics; ATP Synthetase Complexes - metabolism; Bacillus subtilis; Bacillus subtilis - chemistry; Bacillus subtilis - genetics; Bacterial Proteins - analysis; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriophages - genetics; Biological and medical sciences; Cell Membrane - chemistry; Fundamental and applied biological sciences. Psychology; Genes, Reporter; Image Processing, Computer-Assisted; Luminescent Proteins; Membrane Proteins - analysis; Membrane Proteins - metabolism; Microbiology; Microscopy, Fluorescence; Miscellaneous; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; Recombinant Fusion Proteins - metabolism; Succinate Dehydrogenase - analysis; Succinate Dehydrogenase - genetics; Succinate Dehydrogenase - metabolism; Techniques used in virology; Viral Proteins - genetics; Viral Proteins - metabolism; Virology; Reconstruction; Membrane protein; Succinate dehydrogenase; Enzyme; Bacillus subtilis; Bias; Pesticides; Synthase; Bacillaceae; Herbicide; Virus; Bacillales; Sulfonylureas; Bacteria; Oxidoreductases; Localization; Fusion protein; ATP; Phage
• DOI: 10.1099/mic.0.27223-0
• ISSN: 1350-0872

School of Environmental and Life Sciences, Biological Sciences, University of Newcastle, Callaghan, NSW 2308, Australia Correspondence P. J. Lewis Peter.Lewis{at}newcastle.edu.au The subcellular localization of membrane proteins in Bacillus subtilis was examined by using fluorescent protein fusions. ATP synthase and succinate dehydrogenase were found to localize within discrete domains on the membrane rather than being homogeneously distributed around the cell periphery as expected. Dual labelling of cells indicated partial colocalization of ATP synthase and succinate dehydrogenase. Further analysis using an ectopically expressed phage protein gave the same localization patterns as ATP synthase and succinate dehydrogenase, implying that membrane proteins are restricted to domains within the membrane. 3D reconstruction of images of the localization of ATP synthase showed that domains were not regular and there was no bias for localization to cell poles or any other positions. Further analysis revealed that this localization was highly dynamic, but random, implying that integral membrane proteins are free to diffuse two-dimensionally around the cytoplasmic membrane. Abbreviations: FRAP, fluorescence recovery after photobleaching The online version of this paper at http://mic.sgmjournals.org has supplementary movie files and a supplementary figure. Present address: Life Therapeutics, PO Box 6126, Frenchs Forest, NSW 2086, Australia.