Localization and environmental regulation of MCP-like proteins in Rhodobacter sphaeroides.
Chemotaxis to many compounds by Rhodobacter sphaeroides requires transport and at least partial metabolism of the chemoeffector. Previous investigations using phototrophically grown cells have failed to find any homologues of the MCP chemoreceptors identified in Escherichia coli. However, using an antibody raised against the highly conserved domain of E. coli Tsr, MCP-like proteins were identified in R. sphaeroides WS8N. Analysis using Western blotting and immunogold electron microscopy showed that expression of these MCP-like proteins is environmentally regulated and that receptors are targeted to two different cellular locations: the poles of the cells and the cytoplasm. In aerobically grown cells, these proteins were shown by immunoelectron microscopy to localize predominantly to the cell poles and to an electron-dense body in the cytoplasm. Western blot analysis indicated a 17-fold reduction in protein concentration when cells were grown in the light. The number of immunogold particles was also dramatically reduced in anaerobically light-grown cells and their cellular distribution was altered. Fewer receptors localized to the cell poles and more particles randomly distributed within the cell, but the cytoplasmic cluster remained. These trends were more pronounced in cells grown anaerobically under dim light than in those grown anaerobically under bright light, suggesting that expression is controlled by redox state and either light intensity or the extent of photosynthetic membrane synthesis. Recent work on E. coli chemosensing suggests that oligomerization of receptors and chemosensory proteins is important for sensory signalling. The data presented here suggest that this oligomerization can occur with cytoplasmic receptors and also provides an explanation for the multiple copies of chemosensory proteins in R. sphaeroides.
Department of Biochemistry, University of Oxford, UK.
D M
DM
Harrison
J
J
Skidmore
J P
JP
Armitage
J R
JR
Maddock
D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002628:Chemoreceptor Cells; D004926:Escherichia coli / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial; D008027:Light; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology / Q000648:ultrastructure; D000072236:Methyl-Accepting Chemotaxis Proteins; D012242:Rhodobacter sphaeroides / Q000235:genetics / Q000502:physiology*; D013347:Subcellular Fractions
D001426:Bacterial Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; C089936:TlpC protein, Bacillus subtilis; C057635:Tsr protein, Bacteria; C000607604:tsr protein, E coli; C075138:TlpA protein, Salmonella