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Use of Green Fluorescent Protein (GFT)-Chimeras to Study Cytoskeletal Assembly and Dynamics

Published online by Cambridge University Press:  02 July 2020

G. G. Gundersen ,
A. Mikhailov ,
J. L. Martys ,
L. Ho ,
R. K. H. Liem ,
L. Smelinov  and
E.E. Marcantonio
G. G. Gundersen
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
A. Mikhailov
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
J. L. Martys
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
L. Ho
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
R. K. H. Liem
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
L. Smelinov
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
E.E. Marcantonio
Affiliation:
Departments of Cell Biology & Anatomy and Pathology, Columbia University, NY, NY10032
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Extract

The cytoskeleton plays an important role in cell structure, polarity, locomotion and division. Individual elements of the cytoskeleton are composed of subunit proteins which assemble and disassemble in specific places and times within the cell. Knowledge of the temporal and spatial regulation of subunit assembly and disassembly is essential to understanding how the cytoskeleton contributes to cellular activities. The assembly and dynamics of two cytoskeletal structures, namely adhesion plaques (APs) and intermediate filaments (IFs), have been difficult to study by traditional methods. We have generated GFP-chimeras to label these structures and to study their dynamics in motile fibroblasts.

To study the dynamics of APs, we prepared stable 3T3 cell lines expressing a GFP-β1 integrin chimera. The chimera was prepared by fusing GFP to the transmembrane and cytoplasmic portions of β1 intergrin, since previous studies had shown that the cytoplasmic tail of β integrins is sufficient to direct integrins to APs.

Type
Detection and Application of Green (and other Colored) Fluorescent Proteins
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 1010 - 1011
Copyright
Copyright © Microscopy Society of America

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References

l.LaFlamme, S.E. et al J. Cell Biol. 126 (1994) 1287.CrossRefGoogle Scholar
2.Klymkowsky, M.. Nature 291(1981) 249.CrossRefGoogle Scholar
3Gurland, G. and Gundersen, G.G.. J. Cell Biol. 131 (1995) 1275.CrossRefGoogle Scholar