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The Helical MreB Cytoskeleton in Escherichia coli MC1000/pLE7 Is an Artifact of the N-Terminal Yellow Fluorescent Protein Tag

Swulius, Matthew T. and Jensen, Grant J. (2012) The Helical MreB Cytoskeleton in Escherichia coli MC1000/pLE7 Is an Artifact of the N-Terminal Yellow Fluorescent Protein Tag. Journal of Bacteriology, 194 (23). pp. 6382-6386. ISSN 0021-9193. PMCID PMC3497537. doi:10.1128/JB.00505-12. https://resolver.caltech.edu/CaltechAUTHORS:20121219-145126195

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Abstract

Based on fluorescence microscopy, the actin homolog MreB has been thought to form extended helices surrounding the cytoplasm of rod-shaped bacterial cells. The presence of these and other putative helices has come to dominate models of bacterial cell shape regulation, chromosome segregation, polarity, and motility. Here we use electron cryotomography to show that MreB does in fact form extended helices and filaments in Escherichia coli when yellow fluorescent protein (YFP) is fused to its N terminus but native (untagged) MreB expressed to the same levels does not. In contrast, mCherry fused to an internal loop (MreB-RFP^(SW)) does not induce helices. The helices are therefore an artifact of the placement of the fluorescent protein tag. YFP-MreB helices were also clearly distinguishable from the punctate, “patchy” localization patterns of MreB-RFP^(SW), even by standard light microscopy. The many interpretations in the literature of such punctate patterns as helices should therefore be reconsidered.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1128/JB.00505-12 DOIArticle
http://jb.asm.org/content/194/23/6382PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497537/PubMed CentralArticle
ORCID:
AuthorORCID
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2012 American Society for Microbiology. Received 30 March 2012 Accepted 31 July 2012. Published ahead of print 17 August 2012. We thank L. Rothfield for E. coli strains MC1000/pLE7 and MC1000/ pLE6. We also thank Piet De Boer for E. coli strain FB76. This work was supported by NIH grant R01 GM094800B to G.J.J.
Funders:
Funding AgencyGrant Number
NIHR01 GM094800B
Issue or Number:23
PubMed Central ID:PMC3497537
DOI:10.1128/JB.00505-12
Record Number:CaltechAUTHORS:20121219-145126195
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20121219-145126195
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:36059
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:20 Dec 2012 21:38
Last Modified:09 Nov 2021 23:19

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