scholarly journals Intermediate Filaments in Motion: Observations of Intermediate Filaments in Cells Using Green Fluorescent Protein-Vimentin

1999 ◽  
Vol 10 (5) ◽  
pp. 1289-1295 ◽  
Author(s):  
Jayme L. Martys ◽  
Chung-Liang Ho ◽  
Ronald K. H. Liem ◽  
Gregg G. Gundersen
Keyword(s):  
Green Fluorescent Protein ◽  
Intermediate Filaments ◽  
Fluorescent Protein ◽  
Green Fluorescent ◽  
In Cells

Novel features of intermediate filament dynamics revealed by green fluorescent protein chimeras

1998 ◽  
Vol 111 (13) ◽  
pp. 1767-1778 ◽  
Author(s):  
C.L. Ho ◽  
J.L. Martys ◽  
A. Mikhailov ◽  
G.G. Gundersen ◽  
R.K. Liem
Keyword(s):  
Green Fluorescent Protein ◽  
Dynamic Behavior ◽  
Cell Lines ◽  
Intermediate Filaments ◽  
Intermediate Filament ◽  
Fluorescent Protein ◽  
Nih3t3 Cells ◽  
Green Fluorescent ◽  
Filament Network ◽  
Perinuclear Area

In order to study the dynamic behavior of intermediate filament networks in living cells, we have prepared constructs fusing green fluorescent protein to intermediate filament proteins. Vimentin fused to green fluorescent protein labeled the endogenous intermediate filament network. We generated stable SW13 and NIH3T3 cell lines that express an enhanced green fluorescent protein fused to the N-terminus of full-length vimentin. We were able to observe the dynamic behavior of the intermediate filament network in these cells for periods as long as 4 hours (images acquired every 2 minutes). In both cell lines, the vimentin network constantly moves in a wavy manner. In the NIH3T3 cells, we observed extension of individual vimentin filaments at the edge of the cell. This movement is dependent on microtubules, since the addition of nocodazole stopped the extension of the intermediate filaments. Injection of anti-IFA causes the redistribution or ‘collapse’ of intermediate filaments. We injected anti-IFA antibodies into NIH3T3 cells stably expressing green fluorescent protein fused to vimentin and found that individual intermediate filaments move slowly towards the perinuclear area without obvious disassembly. These results demonstrate that individual intermediate filaments are translocated during the collapse, rather than undergoing disassembly-induced redistribution. Injections of tubulin antibodies disrupt the interactions between intermediate filaments and stable microtubules and cause the collapse of the vimentin network showing that these interactions play an important role in keeping the intermediate filament network extended. The nocodazole inhibition of intermediate filament extension and the anti-IFA microinjection experiments are consistent with a model in which intermediate filaments exhibit an extended distribution when tethered to microtubules, but are translocated to the perinuclear area when these connections are severed.

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

1998 ◽  
Vol 4 (S2) ◽  
pp. 1010-1011
Author(s):  
G. G. Gundersen ◽  
A. Mikhailov ◽  
J. L. Martys ◽  
L. Ho ◽  
R. K. H. Liem ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Cell Lines ◽  
Intermediate Filaments ◽  
Fluorescent Protein ◽  
Cell Structure ◽  
Β1 Integrin ◽  
Spatial Regulation ◽  
Temporal And Spatial ◽  
Green Fluorescent ◽  
3T3 Cell Lines

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.

Imaging myosin 10 in cells

2004 ◽  
Vol 32 (5) ◽  
pp. 689-693 ◽  
Author(s):  
D. Tacon ◽  
P.J. Knight ◽  
M. Peckham
Keyword(s):  
Green Fluorescent Protein ◽  
Cell Biology ◽  
Fluorescent Protein ◽  
Major Task ◽  
Green Fluorescent ◽  
In Cells

Cellular motors (kinesin, dynein and myosin) are ubiquitous. A major task in cell biology is to determine how they function in cells. Here we focus on myosin 10, an intrafilopodial motor, and show how imaging green fluorescent protein fused to myosin 10 or its tail domains can help us understand the function of this myosin.

scholarly journals Complexes between Herpes Simplex Virus Glycoproteins gD, gB, and gH Detected in Cells by Complementation of Split Enhanced Green Fluorescent Protein

2007 ◽  
Vol 81 (20) ◽  
pp. 11532-11537 ◽  
Author(s):  
Elisa Avitabile ◽  
Cristina Forghieri ◽  
Gabriella Campadelli-Fiume
Keyword(s):  
Herpes Simplex Virus ◽  
Green Fluorescent Protein ◽  
Herpes Simplex ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Split Egfp ◽  
Green Fluorescent ◽  
Simplex Virus ◽  
In Cells

ABSTRACT The interactions between herpes simplex virus gD and its nectin1 receptor or between gD, gB, and gH were analyzed by complementation of the N and C portions of split enhanced green fluorescent protein (EGFP) fused to the glycoproteins. The gDN-NectC complex was readily detected; the gDN-gCC complex was undetectable, highlighting the specificity of the assay. Split EGFP complementation was detected between proteins designated gDN+gHC, gDN+gBC, and gHN+gBC+wtgD (gB was deleted of endocytosis motifs), both in cells transfected with two-tree glycoproteins and in syncytia. The in situ assay provides evidence that gD interacts with gH and gB independently of each other and supports a model whereby gH and gB in complex exert their activities to gD.

scholarly journals Interaction of a Host Protein with Core Complexes of Bacteriophage Φ6 To Control Transcription

2010 ◽  
Vol 84 (9) ◽  
pp. 4821-4825 ◽  
Author(s):  
Xueying Qiao ◽  
Yang Sun ◽  
Jian Qiao ◽  
Leonard Mindich
Keyword(s):  
Green Fluorescent Protein ◽  
Rna Polymerase ◽  
Fluorescent Protein ◽  
Host Protein ◽  
Genomic Segment ◽  
Double Stranded Rna ◽  
Infection Period ◽  
The Family ◽  
Green Fluorescent ◽  
In Cells

ABSTRACT Bacteriophages of the family Cystoviridae have genomes consisting of three double-stranded RNA (dsRNA) segments, L, S, and M, packaged within a polyhedral capsid along with RNA polymerase. Transcription of genomic segment L is activated by the interaction of host protein YajQ with the capsid structure. Segment L codes for the proteins of the inner capsid, which are expressed early in infection. Green fluorescent protein (GFP) fusions with YajQ produce uniform fluorescence in uninfected cells and in cells infected with viruses not dependent on YajQ. Punctate fluorescence develops when cells are infected with YajQ-dependent viruses. It appears that the host protein binds to the infecting particles and remains with them during the entire infection period.

scholarly journals When a Day Makes a Difference. Interpreting Data from Endoplasmic Reticulum-Targeted Green Fluorescent Protein Fusions in Cells Grown in Suspension Culture

2002 ◽  
Vol 128 (2) ◽  
pp. 341-344 ◽  
Author(s):  
Staffan Persson ◽  
John Love ◽  
Pei-Lan Tsou ◽  
Dominique Robertson ◽  
William F. Thompson ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Green Fluorescent Protein ◽  
Suspension Culture ◽  
Fluorescent Protein ◽  
Green Fluorescent ◽  
In Cells

Microinjection reveals cell-to-cell movement of green fluorescent protein in cells of maize coleoptiles

Planta ◽  
2001 ◽  
Vol 212 (5-6) ◽  
pp. 692-695 ◽  
Author(s):  
C. L. Wymer ◽  
J. M. Fernández-ábalos ◽  
J. H. Doonan
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Cell Movement ◽  
Green Fluorescent ◽  
In Cells

Sex differences in cells expressing green fluorescent protein under the control of the estrogen receptor-α promoter in the hypothalamus of mice

2015 ◽  
Vol 101 ◽  
pp. 44-52 ◽  
Author(s):  
Chaw Kyi-Tha-Thu ◽  
Kota Okoshi ◽  
Hiroto Ito ◽  
Ken-Ichi Matsuda ◽  
Mitsuhiro Kawata ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Sex Differences ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Estrogen Receptor Α ◽  
Green Fluorescent ◽  
In Cells
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