REL
2017-07-04
2018-01-24
2018-01-24
Tilt-series of e. coli carrying the ple7 plasmid carrying YFP-MreB hyper-overexpressed by induction with 1uM IPTG
Matthew
Swulius
Division of Biology, California Institute of Technology
1200 E California Blvd
Pasadena
California
United States
91125
Grant
Jensen
Division of Biology, California Institute of Technology
1200 E California Blvd
Pasadena
California
United States
91125
Swulius MT
Jensen GJ
5.0
10.6019/EMPIAR-10115
EMDB
EMD-3812
Swulius MT
Jensen GJ
The helical MreB cytoskeleton in Escherichia coli MC1000/pLE7 is an artifact of the N-Terminal yellow fluorescent protein tag
Journal of bacteriology
J. Bacteriol.
23
194
6382
6386
2012
English
10.1128/jb.00505-12
22904287
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. In the publication electron cryotomography was used 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.
Tilt-series for e. coli carrying the ple7 plasmid carrying YFP-MreB hyper-overexpressed by induction with 1uM IPTG
/data
tilt series
MRC
MRC
5
130
1
130
SIGNED 16 BIT INTEGER
2032
variable
2032
variable