Screen for localized proteins in Caulobacter crescentus.
Precise localization of individual proteins is required for processes such as motility, chemotaxis, cell-cycle progression, and cell division in bacteria, but the number of proteins that are localized in bacterial species is not known. A screen based on transposon mutagenesis and fluorescence activated cell sorting was devised to identify large numbers of localized proteins, and employed in Caulobacter crescentus. From a sample of the clones isolated in the screen, eleven proteins with no previously characterized localization in C. crescentus were identified, including six hypothetical proteins. The localized hypothetical proteins included one protein that was localized in a helix-like structure, and two proteins for which the localization changed as a function of the cell cycle, suggesting that complex three-dimensional patterns and cell cycle-dependent localization are likely to be common in bacteria. Other mutants produced localized fusion proteins even though the transposon has inserted near the 5' end of a gene, demonstrating that short peptides can contain sufficient information to localize bacterial proteins. The screen described here could be used in most bacterial species.
Russell
Jay H
JH
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania, United States of America.
Keiler
Kenneth C
KC
D001426:Bacterial Proteins / Q000032:analysis*; D001483:Base Sequence; D016935:Caulobacter crescentus / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D018274:Electroporation; D049452:Green Fluorescent Proteins / Q000235:genetics; D016296:Mutagenesis; D010957:Plasmids
D001426:Bacterial Proteins; D017931:DNA Primers; D049452:Green Fluorescent Proteins