Localization of components of the chemotaxis machinery of Escherichia coli using fluorescent protein fusions. Export

@article{2000Sourjik, abstract = {We prepared fusions of yellow fluorescent protein [the YFP variant of green fluorescent protein (GFP)] with the cytoplasmic chemotaxis proteins CheY, CheZ and CheA and the flagellar motor protein FliM, and studied their localization in wild-type and mutant cells of Escherichia coli. All but the CheA fusions were functional. The cytoplasmic proteins CheY, CheZ and CheA tended to cluster at the cell poles in a manner similar to that observed earlier for methyl-accepting chemotaxis proteins (MCPs), but only if MCPs were present. Co-localization of CheY and CheZ with MCPs was CheA dependent, and co-localization of CheA with MCPs was CheW dependent, as expected. Co-localization with MCPs was confirmed by immunofluorescence using an anti-MCP primary antibody. The motor protein FliM appeared as discrete spots on the sides of the cell. These were seen in wild-type cells and in a fliN mutant, but not in flhC or fliG mutants. Co-localization with flagellar structures was confirmed by immunofluorescence using an antihook primary antibody. Surprisingly, we did not observe co-localization of CheY with motors, even under conditions in which cells tumbled.}, address = {Department of Molecular and Cellular Biology, The Biological Laboratories, Harvard University, Cambridge, MA 02138, USA.}, author = {Sourjik, V. and Berg, H. C.}, citeulike-article-id = {2531915}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/10972797}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=10972797}, issn = {0950-382X}, journal = {Mol Microbiol}, keywords = {chemotaxis, fluorescent, fusions, protein}, month = {August}, number = {4}, pages = {740--751}, posted-at = {2008-03-14 12:47:36}, priority = {2}, title = {Localization of components of the chemotaxis machinery of Escherichia coli using fluorescent protein fusions.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/10972797}, volume = {37}, year = {2000} }

TY - JOUR ID - 2000Sourjik L3 - citeulike-article-id:2531915 N2 - We prepared fusions of yellow fluorescent protein [the YFP variant of green fluorescent protein (GFP)] with the cytoplasmic chemotaxis proteins CheY, CheZ and CheA and the flagellar motor protein FliM, and studied their localization in wild-type and mutant cells of Escherichia coli. All but the CheA fusions were functional. The cytoplasmic proteins CheY, CheZ and CheA tended to cluster at the cell poles in a manner similar to that observed earlier for methyl-accepting chemotaxis proteins (MCPs), but only if MCPs were present. Co-localization of CheY and CheZ with MCPs was CheA dependent, and co-localization of CheA with MCPs was CheW dependent, as expected. Co-localization with MCPs was confirmed by immunofluorescence using an anti-MCP primary antibody. The motor protein FliM appeared as discrete spots on the sides of the cell. These were seen in wild-type cells and in a fliN mutant, but not in flhC or fliG mutants. Co-localization with flagellar structures was confirmed by immunofluorescence using an antihook primary antibody. Surprisingly, we did not observe co-localization of CheY with motors, even under conditions in which cells tumbled. IS - 4 JF - Mol Microbiol SN - 0950-382X AD - Department of Molecular and Cellular Biology, The Biological Laboratories, Harvard University, Cambridge, MA 02138, USA. EP - 751 TI - Localization of components of the chemotaxis machinery of Escherichia coli using fluorescent protein fusions. VL - 37 SP - 740 KW - chemotaxis KW - fluorescent KW - fusions KW - protein AU - Sourjik, V AU - Berg, HC PY - 2000/08// UR - http://view.ncbi.nlm.nih.gov/pubmed/10972797 ER -