Journal of Bacteriology, September 2005, p. 6187-6196, Vol. 187, No. 17
0021-9193/05/$08.00+0 doi:10.1128/JB.187.17.6187-6196.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
Presence of Multiple Sites Containing Polar Material in Spherical Escherichia coli Cells That Lack MreB
Trine Nilsen,
Arthur W. Yan,
Gregory Gale, and
Marcia B. Goldberg*
Infectious Disease Division, Massachusetts General Hospital, Cambridge, Massachusetts
Received 10 November 2004/
Accepted 15 June 2005
In rod-shaped bacteria, certain proteins are specifically localized to the cell poles. The nature of the positional information that leads to the proper localization of these proteins is unclear. In a screen for factors required for the localization of the Shigella sp. actin assembly protein IcsA to the bacterial pole, a mutant carrying a transposon insertion in mreB displayed altered targeting of IcsA. The phenotype of cells containing a transposon insertion in mreB was indistinguishable from that of cells containing a nonpolar mutation in mreB or that of wild-type cells treated with the MreB inhibitor A22. In cells lacking MreB, a green fluorescent protein (GFP) fusion to a cytoplasmic derivative of IcsA localized to multiple sites. Secreted full-length native IcsA was present in multiple faint patches on the surfaces of these cells in a pattern similar to that seen for the cytoplasmic IcsA-GFP fusion. EpsM, the polar Vibrio cholerae inner membrane protein, also localized to multiple sites in mreB cells and colocalized with IcsA, indicating that localization to multiple sites is not unique to IcsA. Our results are consistent with the requirement, either direct or indirect, for MreB in the restriction of certain polar material to defined sites within the cell and, in the absence of MreB, with the formation of ectopic sites containing polar material.
* Corresponding author. Mailing address: Bacterial Pathogenesis Laboratories, University Park, 65 Landsdowne Street, Cambridge, MA 02139. Phone: (617) 768-8740. Fax: (617) 768-8738. E-mail: mgoldberg1{at}partners.org.
Present address: US Genomics, Woburn, MA.
Present address: Tufts Medical School, Boston, MA.
Journal of Bacteriology, September 2005, p. 6187-6196, Vol. 187, No. 17
0021-9193/05/$08.00+0 doi:10.1128/JB.187.17.6187-6196.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
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Copyright © 2005 by the American Society for Microbiology. All rights reserved.