The helical mreb cytoskeleton in Escherichia coli MC1000/pLE7 is an artifact of the N-terminal yellow fluorescent protein tag

Matthew T. Swulius, Grant J. Jensen

Research output: Contribution to journalArticlepeer-review

133 Scopus citations

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 (MreBRFPSW) 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-RFPSW, even by standard light microscopy. The many interpretations in the literature of such punctate patterns as helices should therefore be reconsidered.

Original languageEnglish (US)
Pages (from-to)6382-6386
Number of pages5
JournalJournal of bacteriology
Volume194
Issue number23
DOIs
StatePublished - Dec 2012

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

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