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Journal of Bacteriology, October 2005, p. 6874-6882, Vol. 187, No. 20
0021-9193/05/$08.00+0     doi:10.1128/JB.187.20.6874-6882.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Distinct Constrictive Processes, Separated in Time and Space, Divide Caulobacter Inner and Outer Membranes{dagger}

Ellen M. Judd,1,3,{ddagger} Luis R. Comolli,4,{ddagger} Joseph C. Chen,3 Kenneth H. Downing,4 W. E. Moerner,2 and Harley H. McAdams3*

Departments of Applied Physics,1 Chemistry, Stanford University,2 Department of Developmental Biology, Stanford University School of Medicine, Stanford, California 94305,3 Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 947204

Received 18 April 2005/ Accepted 25 July 2005

Cryoelectron microscope tomography (cryoEM) and a fluorescence loss in photobleaching (FLIP) assay were used to characterize progression of the terminal stages of Caulobacter crescentus cell division. Tomographic cryoEM images of the cell division site show separate constrictive processes closing first the inner membrane (IM) and then the outer membrane (OM) in a manner distinctly different from that of septum-forming bacteria. FLIP experiments had previously shown cytoplasmic compartmentalization (when cytoplasmic proteins can no longer diffuse between the two nascent progeny cell compartments) occurring 18 min before daughter cell separation in a 135-min cell cycle so the two constrictive processes are separated in both time and space. In the very latest stages of both IM and OM constriction, short membrane tether structures are observed. The smallest observed prefission tethers were 60 nm in diameter for both the inner and outer membranes. Here, we also used FLIP experiments to show that both membrane-bound and periplasmic fluorescent proteins diffuse freely through the FtsZ ring during most of the constriction procession.

* Corresponding author. Mailing address: Department of Developmental Biology, Stanford University School of Medicine, 279 Campus Drive, Beckman Center B300, Stanford, CA 94305-5329. Phone: (650) 858-1864. Fax: (650) 725-7739. E-mail: hmcadams{at}stanford.edu.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} Equal contributions.

Journal of Bacteriology, October 2005, p. 6874-6882, Vol. 187, No. 20
0021-9193/05/$08.00+0     doi:10.1128/JB.187.20.6874-6882.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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