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

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.

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Mammalian cell lines expressing functional RNA polymerase II tagged with the green fluorescent protein

Journal of Cell Science ◽

10.1242/jcs.113.15.2679 ◽

2000 ◽

Vol 113 (15) ◽

pp. 2679-2683 ◽

Cited By ~ 3

Author(s):

K. Sugaya ◽

M. Vigneron ◽

P.R. Cook

Keyword(s):

Green Fluorescent Protein ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Cell Lines ◽

Fluorescent Protein ◽

Living Cells ◽

Temperature Sensitive ◽

Mammalian Cell Lines ◽

Eukaryotic Genes ◽

Green Fluorescent

RNA polymerase II is a multi-subunit enzyme responsible for transcription of most eukaryotic genes. It associates with other complexes to form enormous multifunctional ‘holoenzymes’ involved in splicing and polyadenylation. We wished to study these different complexes in living cells, so we generated cell lines expressing the largest, catalytic, subunit of the polymerase tagged with the green fluorescent protein. The tagged enzyme complements a deficiency in tsTM4 cells that have a temperature-sensitive mutation in the largest subunit. Some of the tagged subunit is incorporated into engaged transcription complexes like the wild-type protein; it both resists extraction with sarkosyl and is hyperphosphorylated at its C terminus. Remarkably, subunits bearing such a tag can be incorporated into the active enzyme, despite the size and complexity of the polymerizing complex. Therefore, these cells should prove useful in the analysis of the dynamics of transcription in living cells.

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Imaging myosin 10 in cells

Biochemical Society Transactions ◽

10.1042/bst0320689 ◽

2004 ◽

Vol 32 (5) ◽

pp. 689-693 ◽

Cited By ~ 10

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.

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Non-Target Effects of Green Fluorescent Protein (GFP)-Derived Double-Stranded RNA (dsRNA-GFP) Used in Honey Bee RNA Interference (RNAi) Assays

Insects ◽

10.3390/insects4010090 ◽

2013 ◽

Vol 4 (1) ◽

pp. 90-103 ◽

Cited By ~ 43

Author(s):

Francis Nunes ◽

Aline Aleixo ◽

Angel Barchuk ◽

Ana Bomtorin ◽

Christina Grozinger ◽

...

Keyword(s):

Green Fluorescent Protein ◽

Rna Interference ◽

Honey Bee ◽

Fluorescent Protein ◽

Double Stranded Rna ◽

Green Fluorescent ◽

Target Effects

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

Journal of Virology ◽

10.1128/jvi.01343-07 ◽

2007 ◽

Vol 81 (20) ◽

pp. 11532-11537 ◽

Cited By ~ 83

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.

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Oligonucleotide-mediated gene editing is underestimated in cells expressing mutated green fluorescent protein and is positively associated with target protein expression

The Journal of Gene Medicine ◽

10.1002/jgm.1639 ◽

2012 ◽

Vol 14 (2) ◽

pp. 109-119 ◽

Cited By ~ 2

Author(s):

Petra Disterer ◽

Ioannis Papaioannou ◽

Vanessa C. Evans ◽

J. Paul Simons ◽

James S. Owen

Keyword(s):

Green Fluorescent Protein ◽

Protein Expression ◽

Gene Editing ◽

Fluorescent Protein ◽

Target Protein ◽

Green Fluorescent ◽

In Cells

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

Molecular Biology of the Cell ◽

10.1091/mbc.10.5.1289 ◽

1999 ◽

Vol 10 (5) ◽

pp. 1289-1295 ◽

Cited By ~ 50

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

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

PLANT PHYSIOLOGY ◽

10.1104/pp.010840 ◽

2002 ◽

Vol 128 (2) ◽

pp. 341-344 ◽

Cited By ~ 4

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

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Comparing Avocado, Swamp Bay, and Camphortree as Hosts of Raffaelea lauricola Using a Green Fluorescent Protein (GFP)-Labeled Strain of the Pathogen

Phytopathology ◽

10.1094/phyto-02-16-0072-r ◽

2017 ◽

Vol 107 (1) ◽

pp. 70-74 ◽

Cited By ~ 6

Author(s):

A. S. Campbell ◽

R. C. Ploetz ◽

J. A. Rollins

Keyword(s):

Green Fluorescent Protein ◽

Fluorescent Protein ◽

Severe Disease ◽

Persea Americana ◽

Laurel Wilt ◽

Xyleborus Glabratus ◽

Raffaelea Lauricola ◽

The Family ◽

Pathogen Colonization ◽

Green Fluorescent

Raffaelea lauricola, a fungal symbiont of the ambrosia beetle Xyleborus glabratus, causes laurel wilt in members of the Lauraceae plant family. North American species in the family, such as avocado (Persea americana) and swamp bay (P. palustris), are particularly susceptible to laurel wilt, whereas the Asian camphortree (Cinnamomum camphora) is relatively tolerant. To determine whether susceptibility is related to pathogen colonization, a green fluorescent protein-labeled strain of R. lauricola was generated and used to inoculate avocado, swamp bay, and camphortree. Trees were harvested 3, 10, and 30 days after inoculation (DAI), and disease severity was rated on a 1-to-10 scale. By 30 DAI, avocado and swamp bay developed significantly more severe disease than camphortree (mean severities of 6.8 and 5.5 versus 1.6, P < 0.003). The extent of xylem colonization was recorded as the percentage of lumena that were colonized by the pathogen. More xylem was colonized in avocado than camphortree (0.9% versus 0.1%, P < 0.03) but colonization in swamp bay (0.4%) did not differ significantly from either host. Although there were significant correlations between xylem colonization and laurel wilt severity in avocado (r = 0.74), swamp bay (r = 0.82), and camphortree (r = 0.87), even severely affected trees of all species were scarcely colonized by the pathogen.

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A photoactivatable green-fluorescent protein from the phylum Ctenophora

Proceedings of The Royal Society B Biological Sciences ◽

10.1098/rspb.2009.1774 ◽

2009 ◽

Vol 277 (1685) ◽

pp. 1155-1160 ◽

Cited By ~ 25

Author(s):

Steven H. D. Haddock ◽

Nadia Mastroianni ◽

Lynne M. Christianson

Keyword(s):

Green Fluorescent Protein ◽

Fluorescent Protein ◽

Fluorescent Proteins ◽

Green Fluorescent Proteins ◽

Optical Probes ◽

Fluorescent Emission ◽

The Family ◽

Fluorescent State ◽

Green Fluorescent

Genes for the family of green-fluorescent proteins (GFPs) have been found in more than 100 species of animals, with some species containing six or more copies producing a variety of colours. Thus far, however, these species have all been within three phyla: Cnidaria, Arthropoda and Chordata. We have discovered GFP-type fluorescent proteins in the phylum Ctenophora, the comb jellies. The ctenophore proteins share the x YG chromophore motif of all other characterized GFP-type proteins. These proteins exhibit the uncommon property of reversible photoactivation, in which fluorescent emission becomes brighter upon exposure to light, then gradually decays to a non-fluorescent state. In addition to providing potentially useful optical probes with novel properties, finding a fluorescent protein in one of the earliest diverging metazoans adds further support to the possibility that these genes are likely to occur throughout animals.

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