Reconstitution of Contractile FtsZ Rings in Liposomes
Masaki Osawa, David E. Anderson, Harold P. Erickson
Supporting Online Material
This supplement contains:
Materials and Methods
Fig. S1
References
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Correction (30 April 2008):
The authors have added further detail to the paragraph
describing purification methods in the Methods section.
The original version can be found
here
This file is in Adobe Acrobat PDF format.
Other Supporting Online Material for this manuscript includes the following:
(available at
www.sciencemag.org/cgi/content/full/1154520/DC1)
Movies S1 to S3
Movie s1
Four time frames are shown from movie S1, with lines drawn to
show the coincidence of constrictions and Z rings. At 0 sec there are multiple dim Z
rings, and no visible constrictions. At 100 sec there are a couple of barely perceptible
indentations (not marked). At 200 and 350 sec, dim Z rings have coalesced into brighter
ones. Lines were drawn from the Z rings to the DIC image in the upper panel. The
constrictions appear to be centered on the Z rings.
Movie s2
The complete time series from which Fig. 4 was extracted.
Times are marked in sec, starting at the first observation about 10 min after mixing
liposomes, FtsZ-mts and GTP. The bright Z ring on the left (arrow) maintained a fairly
deep constriction for the first 430 sec. At 440 and 450 sec it relaxed and expanded a bit,
and at 460 sec it abruptly relaxed completely, letting the liposome wall retract to a
smooth, unconstricted cylinder. A second Z ring on the right (arrow) abruptly relaxed
two min later.
Movie s3
In this experiment the slide was maintained at 37° C for
observation. GTP was 400 μM, as in Movie S1, but it is hydrolyzed 2-3 times faster at the
higher temperature. The movie shows some constrictions already present in the first
frame, which was about 40 min after sample preparation. From 0-600 sec there is some
coalescence of rings, and most of the constrictions deepen, even for rings that did not
grow by coalescence. Constrictions were maximally deep from 600-700 sec. From 800-
1200 sec the rings became dimmer and the constrictions relaxed substantially. Some
constrictions are still visible at 1800 sec, but are much shallower than at 700 sec. The
total width of the movie is 85.5 μm and the inner diameter of the liposome is
approximately 0.3 – 1 μm.
To view these movies, download a QuickTime viewer.
