Absolute Arrangement of Subunits in Cytoskeletal Septin Filaments in Cells Measured by Fluorescence Microscopy.
We resolved the organization of subunits in cytoskeletal polymers in cells by light microscopy. Septin GTPases form linear complexes of about 32 nm length that polymerize into filaments. We visualized both termini of septin complexes by single molecule microscopy in vitro. Complexes appeared as 32 nm spaced localization pairs, and filaments appeared as stretches of equidistant localizations. Cellular septins were resolved as localization pairs and thin stretches of equidistant localizations.
†Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
Charlotte
C
Kaplan
†Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
Bo
B
Jing
†Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
Christian M
CM
Winterflood
‡Life Science Center, Dartmouth College, Hanover, New Hampshire 03755, United States.
Andrew A
AA
Bridges
‡Life Science Center, Dartmouth College, Hanover, New Hampshire 03755, United States.
Patricia
P
Occhipinti
Jürgen
J
Schmied
∥Institute of Molecular Genetics and Cell Biology, University of Ulm, 89081 Ulm, Germany.
Sören
S
Grinhagens
∥Institute of Molecular Genetics and Cell Biology, University of Ulm, 89081 Ulm, Germany.
Thomas
T
Gronemeyer
Philip
P
Tinnefeld
‡Life Science Center, Dartmouth College, Hanover, New Hampshire 03755, United States.
Amy S
AS
Gladfelter
†Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
Jonas
J
Ries
†Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
Helge
H
Ewers
D003599:Cytoskeleton / Q000737:chemistry* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D058112:Septins / Q000737:chemistry*
D029701:Saccharomyces cerevisiae Proteins; D058112:Septins
DNA origami; Septin; cytoskeleton; single molecule; superresolution microscopy