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A tumorigenic actin mutant alters fibroblast morphology and multicellular assembly properties
Cytoskeleton (Hoboken, N.J.), 2013-10, Vol.70 (10), p.635-650
Blache, Ulrich
Silván, Unai
Plodinec, Marija
Suetterlin, Rosmarie
Jakob, Roman
Klebba, Ina
Bentires-Alj, Mohamed
Aebi, Ueli
Schoenenberger, Cora-Ann
2013
Details
Autor(en) / Beteiligte
Blache, Ulrich
Silván, Unai
Plodinec, Marija
Suetterlin, Rosmarie
Jakob, Roman
Klebba, Ina
Bentires-Alj, Mohamed
Aebi, Ueli
Schoenenberger, Cora-Ann
Titel
A tumorigenic actin mutant alters fibroblast morphology and multicellular assembly properties
Ist Teil von
Cytoskeleton (Hoboken, N.J.), 2013-10, Vol.70 (10), p.635-650
Ort / Verlag
United States: Blackwell Publishing Ltd
Erscheinungsjahr
2013
Link zum Volltext
Quelle
MEDLINE
Beschreibungen/Notizen
Tumor initiation and progression are accompanied by complex changes in the cytoarchitecture that at the cellular level involve remodeling of the cytoskeleton. We report on the impact of a mutant β‐actin (G245D‐actin) on cell structure and multicellular assembly properties. To appreciate the effects of the Gly245Asp substitution on the organization of the actin cytoskeleton, we examined the polymerization properties of G245D‐actin in vitro by pyrene polymerization assays and total internal reflection fluorescence microscopy (TIRF). The mutant actin on its own has a significantly reduced polymerization efficiency compared to native actin but also modifies the polymerization of actin in copolymerization experiments. Comparison of the structure of Rat‐2 fibroblasts and a stably transfected derivate called Rat‐2‐sm9 revealed the effects of G245D‐actin in a cellular environment. The overall actin levels in Rat‐2‐sm9 show a 1.6‐fold increase with similar amounts of mutant and wild‐type actin. G245D‐actin expression renders Rat‐2‐sm9 cells highly tumorigenic in nude mice. In Rat‐2‐sm9 monolayers, G245D‐actin triggers the formation of extensive membrane ruffles, which is a characteristic feature of many transformed cells. To approximate complex cell–cell and cell‐matrix interactions that occur in tumors and might modulate the effects of G245D‐actin, we extended our studies to scaffold‐free 3D spheroid cultures. Bright field and scanning electron microscopy (SEM) show that Rat‐2‐sm9 and Rat‐2 cells share essential features of spheroid formation and compaction. However, the resulting spheroids exhibit distinct phenotypes that differ mainly in surface structure and size. The systematic comparison of transformed and normal spheroids by SEM provides new insights into scaffold‐free fibroblast spheroid formation. © 2013 Wiley Periodicals, Inc.
Sprache
Englisch
Identifikatoren
ISSN: 1949-3584
eISSN: 1949-3592
DOI: 10.1002/cm.21120
Titel-ID: cdi_proquest_miscellaneous_1447107016
Format
–
Schlagworte
Actin Cytoskeleton - metabolism
,
actin mutation
,
actin polymerization
,
Actins - metabolism
,
Animals
,
Carcinogenesis - metabolism
,
Carcinogenesis - pathology
,
Cell Proliferation
,
Cell Shape
,
Fibroblasts - metabolism
,
Fibroblasts - pathology
,
Fibroblasts - ultrastructure
,
membrane ruffling
,
Mice
,
Mice, Nude
,
multicellular spheroids
,
Mutant Proteins - metabolism
,
Mutation - genetics
,
Polymerization
,
Rabbits
,
Rats
,
Spheroids, Cellular - metabolism
,
Spheroids, Cellular - pathology
,
Spheroids, Cellular - ultrastructure
,
Stress Fibers - metabolism
,
transformation
,
tumorigenic
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