scholarly journals The N-Terminal Domain of PMTV TGB1 Movement Protein Is Required for Nucleolar Localization, Microtubule Association, and Long-Distance Movement

2010 ◽  
Vol 23 (11) ◽  
pp. 1486-1497 ◽  
Author(s):  
Kathryn M. Wright ◽  
Graham H. Cowan ◽  
Nina I. Lukhovitskaya ◽  
Jens Tilsner ◽  
Alison G. Roberts ◽  
...  
Keyword(s):  
Amino Acids ◽  
Fluorescent Protein ◽  
Movement Protein ◽  
Leading Edge ◽  
Epidermal Cells ◽  
Viral Rna ◽  
Nucleolar Localization ◽  
Long Distance ◽  
Long Distance Movement ◽  
In Cells

The triple-gene-block (TGB)1 protein of Potato mop-top virus (PMTV) was fused to fluorescent proteins and expressed in epidermal cells of Nicotiana benthamiana under the control of the 35S promoter. TGB1 fluorescence was observed in the cytoplasm, nucleus, and nucleolus and occasionally associated with microtubules. When expressed from a modified virus (PMTV.YFP-TGB1) which formed local lesions but was not competent for systemic movement, yellow fluorescent protein (YFP)-TGB1 labeled plasmodesmata in cells at the leading edge of the lesion and plasmodesmata, microtubules, nuclei, and nucleoli in cells immediately behind the leading edge. Deletion of 84 amino acids from the N-terminus of unlabeled TGB1 within the PMTV genome abolished movement of viral RNA to noninoculated leaves. When the same deletion was introduced into PMTV.YFP-TGB1, labeling of microtubules and nucleoli was abolished. The N-terminal 84 amino acids of TGB1 were fused to green fluorescent protein (GFP) and expressed in epidermal cells where GFP localized strongly to the nucleolus (not seen with unfused GFP), indicating that these amino acids contain a nucleolar localization signal; the fusion protein did not label microtubules. This is the first report of nucleolar and microtubule association of a TGB movement protein. The results suggest that PMTV TGB1 requires interaction with nuclear components and, possibly, microtubules for long-distance movement of viral RNA.

scholarly journals Cellular Targets of Functional and Dysfunctional Mutants of Tobacco Mosaic Virus Movement Protein Fused to Green Fluorescent Protein

2000 ◽  
Vol 74 (23) ◽  
pp. 11339-11346 ◽  
Author(s):  
Vitaly Boyko ◽  
Jessica van der Laak ◽  
Jacqueline Ferralli ◽  
Elena Suslova ◽  
Myoung-Ok Kwon ◽  
...  
Keyword(s):  
Amino Acids ◽  
Green Fluorescent Protein ◽  
Tobacco Mosaic Virus ◽  
Inclusion Bodies ◽  
Fluorescent Protein ◽  
Movement Protein ◽  
Leading Edge ◽  
Intercellular Transport ◽  
C Terminus ◽  
Green Fluorescent

ABSTRACT Intercellular transport of tobacco mosaic virus (TMV) RNA involves the accumulation of virus-encoded movement protein (MP) in plasmodesmata (Pd), in endoplasmic reticulum (ER)-derived inclusion bodies, and on microtubules. The functional significance of these interactions in viral RNA (vRNA) movement was tested in planta and in protoplasts with TMV derivatives expressing N- and C-terminal deletion mutants of MP fused to the green fluorescent protein. Deletion of 55 amino acids from the C terminus of MP did not interfere with the vRNA transport function of MP:GFP but abolished its accumulation in inclusion bodies, indicating that accumulation of MP at these ER-derived sites is not a requirement for function in vRNA intercellular movement. Deletion of 66 amino acids from the C terminus of MP inactivated the protein, and viral infection occurred only upon complementation in plants transgenic for MP. The functional deficiency of the mutant protein correlated with its inability to associate with microtubules and, independently, with its absence from Pd at the leading edge of infection. Inactivation of MP by N-terminal deletions was correlated with the inability of the protein to target Pd throughout the infection site, whereas its associations with microtubules and inclusion bodies were unaffected. The observations support a role of MP-interacting microtubules in TMV RNA movement and indicate that MP targets microtubules and Pd by independent mechanisms. Moreover, accumulation of MP in Pd late in infection is insufficient to support viral movement, confirming that intercellular transport of vRNA relies on the presence of MP in Pd at the leading edge of infection.

scholarly journals Temperature-Dependent Wsm1 and Wsm2 Gene-Specific Blockage of Viral Long-Distance Transport Provides Resistance to Wheat streak mosaic virus and Triticum mosaic virus in Wheat

2016 ◽  
Vol 29 (9) ◽  
pp. 724-738 ◽  
Author(s):  
Satyanarayana Tatineni ◽  
Everlyne N. Wosula ◽  
Melissa Bartels ◽  
Gary L. Hein ◽  
Robert A. Graybosch
Keyword(s):  
Mosaic Virus ◽  
Fluorescent Protein ◽  
Wheat Streak Mosaic Virus ◽  
Viral Resistance ◽  
Wheat Cultivars ◽  
Temperature Dependent ◽  
Long Distance ◽  
Long Distance Transport ◽  
Long Distance Movement ◽  
Distance Transport

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) are economically important viral pathogens of wheat. Wheat cvs. Mace, carrying the Wsm1 gene, is resistant to WSMV and TriMV, and Snowmass, with Wsm2, is resistant to WSMV. Viral resistance in both cultivars is temperature sensitive and is effective at 18°C or below but not at higher temperatures. The underlying mechanisms of viral resistance of Wsm1 and Wsm2, nonallelic single dominant genes, are not known. In this study, we found that fluorescent protein–tagged WSMV and TriMV elicited foci that were approximately similar in number and size at 18 and 24°C, on inoculated leaves of resistant and susceptible wheat cultivars. These data suggest that resistant wheat cultivars at 18°C facilitated efficient cell-to-cell movement. Additionally, WSMV and TriMV efficiently replicated in inoculated leaves of resistant wheat cultivars at 18°C but failed to establish systemic infection, suggesting that Wsm1- and Wsm2-mediated resistance debilitated viral long-distance transport. Furthermore, we found that neither virus was able to enter the leaf sheaths of inoculated leaves or crowns of resistant wheat cultivars at 18°C but both were able to do so at 24°C. Thus, wheat cvs. Mace and Snowmass provide resistance at the long-distance movement stage by specifically blocking virus entry into the vasculature. Taken together, these data suggest that both Wsm1 and Wsm2 genes similarly confer virus resistance by temperature-dependent impairment of viral long-distance movement.

scholarly journals The Rate of Cell-to-Cell Movement in Squash of Cucumber Mosaic Virus Is Affected by Sequences of the Capsid Protein

1999 ◽  
Vol 12 (7) ◽  
pp. 628-632 ◽  
Author(s):  
Sek-Man Wong ◽  
Sharon Swee-Chin Thio ◽  
Michael H. Shintaku ◽  
Peter Palukaitis
Keyword(s):  
Amino Acids ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Capsid Protein ◽  
Cell Movement ◽  
Systemic Infection ◽  
Long Distance ◽  
Local Movement ◽  
Long Distance Movement

The M strain of cucumber mosaic virus (CMV) does not infect squash plants systemically and moves very slowly in inoculated cotyledons. Systemic infection and an increase in the rate of local movement were observed when amino acids 129 or 214 of the M-CMV capsid protein (CP) were altered to those present in the Fny strain of CMV. While the opposite alterations to the CP of Fny-CMV inhibited systemic infection of squash, they did not show the same effects on the rates of both cell-to-cell and long-distance movement. However, the ability of CMV to infect squash systemically was affected by the rate of cell-to-cell movement.

scholarly journals The Benyvirus RNA Silencing Suppressor Is Essential for Long-Distance Movement, Requires Both Zinc-Finger and NoLS Basic Residues but Not a Nucleolar Localization for Its Silencing-Suppression Activity

2013 ◽  
Vol 26 (2) ◽  
pp. 168-181 ◽  
Author(s):  
Sotaro Chiba ◽  
Kamal Hleibieh ◽  
Alice Delbianco ◽  
Elodie Klein ◽  
Claudio Ratti ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Rna Silencing ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Nucleolar Localization ◽  
Silencing Suppressor ◽  
Long Distance ◽  
Microscopy Imaging ◽  
Silencing Suppression ◽  
Long Distance Movement

The RNA silencing-suppression properties of Beet necrotic yellow vein virus (BNYVV) and Beet soil-borne mosaic virus (BSBMV) cysteine-rich p14 proteins have been investigated. Suppression of RNA silencing activities were made evident using viral infection of silenced Nicotiana benthamiana 16C, N. benthamiana agroinfiltrated with green fluorescent protein (GFP), and GF-FG hairpin triggers supplemented with viral suppressor of RNA silencing (VSR) constructs or using complementation of a silencing-suppressor-defective BNYVV virus in Chenopodium quinoa. Northern blot analyses of small-interfering RNAs (siRNAs) in agroinfiltration tests revealed reduced amounts of siRNA, especially secondary siRNA, suggesting that benyvirus VSR act downstream of the siRNA production. Using confocal laser-scanning microscopy imaging of infected protoplasts expressing functional p14 protein fused to an enhanced GFP reporter, we showed that benyvirus p14 accumulated in the nucleolus and the cytoplasm independently of other viral factors. Site-directed mutagenesis showed the importance of the nucleolar localization signal embedded in a C4 zinc-finger domain in the VSR function and intrinsic stability of the p14 protein. Conversely, RNA silencing suppression appeared independent of the nucleolar localization of the protein, and a correlation between BNYVV VSR expression and long-distance movement was established.

scholarly journals Inhibition of long-distance movement of RNA silencing signals in Nicotiana benthamiana by Apple chlorotic leaf spot virus 50 kDa movement protein

Virology ◽  
2008 ◽  
Vol 382 (2) ◽  
pp. 199-206 ◽  
Author(s):  
Hajime Yaegashi ◽  
Akihiro Tamura ◽  
Masamichi Isogai ◽  
Nobuyuki Yoshikawa
Keyword(s):  
Leaf Spot ◽  
Nicotiana Benthamiana ◽  
Rna Silencing ◽  
Movement Protein ◽  
Long Distance ◽  
Spot Virus ◽  
Chlorotic Leaf ◽  
Long Distance Movement

scholarly journals Hibiscus Chlorotic Ringspot Virus Coat Protein Is Essential for Cell-to-Cell and Long-Distance Movement but Not for Viral RNA Replication

PLoS ONE ◽  
2014 ◽  
Vol 9 (11) ◽  
pp. e113347 ◽  
Author(s):  
Shengniao Niu ◽  
Francisco M. Gil-Salas ◽  
Sunil Kumar Tewary ◽  
Ashwin Kuppusamy Samales ◽  
John Johnson ◽  
...  
Keyword(s):  
Coat Protein ◽  
Rna Replication ◽  
Ringspot Virus ◽  
Viral Rna ◽  
Long Distance ◽  
Virus Coat Protein ◽  
Virus Coat ◽  
Long Distance Movement

scholarly journals A plant virus-encoded protein facilitates long-distance movement of heterologous viral RNA

1999 ◽  
Vol 96 (4) ◽  
pp. 1212-1217 ◽  
Author(s):  
E. V. Ryabov ◽  
D. J. Robinson ◽  
M. E. Taliansky
Keyword(s):  
Plant Virus ◽  
Viral Rna ◽  
Long Distance ◽  
Long Distance Movement

scholarly journals Endogenous p53 protein generated from wild-type alternatively spliced p53 RNA in mouse epidermal cells

1994 ◽  
Vol 14 (3) ◽  
pp. 1698-1708
Author(s):  
M F Kulesz-Martin ◽  
B Lisafeld ◽  
H Huang ◽  
N D Kisiel ◽  
L Lee
Keyword(s):  
Cell Cycle ◽  
Amino Acids ◽  
Transcriptional Activation ◽  
Cultured Cells ◽  
P53 Protein ◽  
Epidermal Cells ◽  
Wild Type ◽  
Mouse Tissues ◽  
Alternatively Spliced ◽  
In Cells

We previously demonstrated that a wild-type alternatively spliced p53 (p53as) RNA exists in mouse cultured cells and normal mouse tissues at approximately 25 to 33% of the level of the major p53 RNA form. The alternative RNA transcript is 96 nucleotides longer than the major transcript as a result of alternative splicing of intron 10 sequences. The protein expected to be generated from the p53as transcript is 9 amino acids shorter than the major p53 protein and has 17 different amino acids at the carboxyl terminus. We report here that p53as protein exists in nontransformed and malignant epidermal cells and is localized to the nucleus. In addition, p53as protein is preferentially expressed during the G2 phase of the cell cycle and in cells with greater than G2 DNA content compared with the major p53 protein, which is preferentially expressed in G1. The p53as immunoreactivity is elevated and shifted to the G1 phase of the cell cycle following actinomycin D treatment of nontransformed cells but not malignant cells. In view of the dimerization and tetramerization of p53 protein which may be necessary for its DNA binding and transcriptional activation activities, the presence of p53as protein in cells has important implications for understanding the physiological function(s) of the p53 gene.

scholarly journals Dynamics of Bean Dwarf Mosaic Geminivirus Cell-to-Cell and Long-Distance Movement in Phaseolus vulgaris Revealed, Using the Green Fluorescent Protein

1998 ◽  
Vol 11 (4) ◽  
pp. 277-291 ◽  
Author(s):  
M. R. Sudarshana ◽  
H. L. Wang ◽  
W. J. Lucas ◽  
R. L. Gilbertson
Keyword(s):  
Green Fluorescent Protein ◽  
Phaseolus Vulgaris ◽  
Particle Bombardment ◽  
Fluorescent Protein ◽  
Fluorescence Analysis ◽  
Reporter System ◽  
Long Distance ◽  
Viral Dna ◽  
Green Fluorescent ◽  
Long Distance Movement

The cell-to-cell and long-distance movement of the bipartite geminivirus, bean dwarf mosaic (BDMV), in Phaseolus vulgaris plants was examined with the noninvasive reporter, the green fluorescent protein (GFP). A modified GFP gene (mGFP4) was inserted into the BDMV DNA-A component in place of the coat protein gene (BDMVA-mGFP4), and particle bombardment was used to introduce viral DNA into bean seedlings (radicle and hypocotyl tissues). Fluorescence analysis of GFP expressed from BDMVA-mGFP4 established that particle bombardment introduced viral DNA only into epidermal cells, and the requirement for the DNA-B-encoded proteins (BV1 and BC1) in the cell-to-cell movement of BDMVA-mGFP4. This GFP reporter system was used to follow the viral infection process from the seedling stage throughout the entire plant life cycle. In inoculated hypocotyls, BDMV moved from cell to cell through the cortex and showed a striking phloem tropism. Upon entry into phloem tissues, BDMV moved rapidly toward the root via the long-distance transport system, and toward the shoot apex by a combination of cell-to-cell and long-distance movement. Analysis of GFP distribution in systemically infected tissues revealed that BDMV was restricted to phloem cells in both roots and stems. In systemically infected primary and trifoliolate leaves, BDMV infected phloem cells associated with all vein orders (first through fifth), and the capacity of BDMV to exit from phloem tissue into nonphloem cells was correlated with the stage of plant development. Finally, fluorescence analysis of GFP in reproductive tissues established that BDMV infected flower, pod, and seed-coat tissues, but was excluded from the embryo.

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