scholarly journals The Capsid Protein of Turnip Crinkle Virus Overcomes Two Separate Defense Barriers To Facilitate Systemic Movement of the Virus in Arabidopsis

2010 ◽  
Vol 84 (15) ◽  
pp. 7793-7802 ◽  
Author(s):  
Mingxia Cao ◽  
Xiaohong Ye ◽  
Kristen Willie ◽  
Junyan Lin ◽  
Xiuchun Zhang ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Viral Genome ◽  
Single Amino Acid ◽  
Silencing Suppressor ◽  
Turnip Crinkle Virus ◽  
Coding Region ◽  
Long Distance ◽  
Virus Particles ◽  
Systemic Movement ◽  
Long Distance Movement

ABSTRACT The capsid protein (CP) of Turnip crinkle virus (TCV) is a multifunctional protein needed for virus assembly, suppression of RNA silencing-based antiviral defense, and long-distance movement in infected plants. In this report, we have examined genetic requirements for the different functions of TCV CP and evaluated the interdependence of these functions. A series of TCV mutants containing alterations in the CP coding region were generated. These alterations range from single-amino-acid substitutions and domain truncations to knockouts of CP translation. The latter category also contained two constructs in which the CP coding region was replaced by either the cDNA of a silencing suppressor of a different virus or that of green fluorescent protein. These mutants were used to infect Arabidopsis plants with diminished antiviral silencing capability (dcl2 dcl3 dcl4 plants). There was a strong correlation between the ability of mutants to reach systemic leaves and the silencing suppressor activity of mutant CP. Virus particles were not essential for entry of the viral genome into vascular bundles in the inoculated leaves in the absence of antiviral silencing. However, virus particles were necessary for egress of the viral genome from the vasculature of systemic leaves. Our experiments demonstrate that TCV CP not only allows the viral genome to access the systemic movement channel through silencing suppression but also ensures its smooth egress by way of assembled virus particles. These results illustrate that efficient long-distance movement of TCV requires both functions afforded by the CP.

scholarly journals Unusual Long-Distance Movement Strategies of Potato mop-top virus RNAs in Nicotiana benthamiana

2009 ◽  
Vol 22 (4) ◽  
pp. 381-390 ◽  
Author(s):  
Lesley Torrance ◽  
Nina I. Lukhovitskaya ◽  
Mikhail V. Schepetilnikov ◽  
Graham H. Cowan ◽  
Angelika Ziegler ◽  
...  
Keyword(s):  
Capsid Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leaf Extracts ◽  
Long Distance ◽  
Particle Assembly ◽  
Virus Particles ◽  
Systemic Movement ◽  
Translational Readthrough ◽  
Potato Mop Top Virus ◽  
Long Distance Movement

The Potato mop-top virus (PMTV) genome encodes replicase, movement, and capsid proteins on three different RNA species that are encapsidated within tubular rod-shaped particles. Previously, we showed that the protein produced on translational readthrough (RT) of the coat protein (CP) gene, CP-RT, is associated with one extremity of the virus particles, and that the two RNAs encoding replicase and movement proteins can move long distance in the absence of the third RNA (RNA-CP) that encodes the capsid proteins, CP and CP-RT. Here, we examined the roles of the CP and CP-RT proteins on RNA movement using infectious clones carrying mutations in the CP and CP-RT coding domains. The results showed that, in infections established with mutant RNA-CP expressing CP together with truncated CP-RT, systemic movement of the mutant RNA-CP was inhibited but not the movement of the other two RNAs. Furthermore, RNA-CP long-distance movement was inhibited in a mutant clone expressing only CP in the absence of the CP-RT polypeptide. CP-RT was not necessary for particle assembly because virions were observed in leaf extracts infected with the CP-RT deletion mutants. RNA-CP moved long distance when protein expression was suppressed completely or when CP expression was suppressed so that only CP-RT or truncated CP-RT was expressed. CP-RT but not CP interacted with the movement protein TGB1 in the yeast two-hybrid system. CP-RT and TGB1 were detected by enzyme-linked immunosorbent assay in virus particles and the long-distance movement of RNA-CP was correlated with expression of CP-RT that interacted with TGB1; mutant RNA-CP expressing truncated CP-RT proteins that did not interact with TGB1 formed virions but did not move to upper noninoculated leaves. The results indicate that PMTV RNA-CP can move long distance in two distinct forms: either as a viral ribonucleoprotein complex or as particles that are most likely associated with CP-RT and TGB1.

scholarly journals Formation of virions is strictly required for turnip yellows virus long-distance movement in plants

2014 ◽  
Vol 95 (2) ◽  
pp. 496-505 ◽  
Author(s):  
Clémence Hipper ◽  
Baptiste Monsion ◽  
Diane Bortolamiol-Bécet ◽  
Véronique Ziegler-Graff ◽  
Véronique Brault
Keyword(s):  
Minor Component ◽  
Long Distance ◽  
Virus Particles ◽  
Systemic Movement ◽  
Long Distance Transport ◽  
Viral Particles ◽  
Turnip Yellows Virus ◽  
In Trans ◽  
Long Distance Movement ◽  
Distance Transport

Viral genomic RNA of the Turnip yellows virus (TuYV; genus Polerovirus; family Luteoviridae) is protected in virions formed by the major capsid protein (CP) and the minor component, the readthrough (RT*) protein. Long-distance transport, used commonly by viruses to systemically infect host plants, occurs in phloem sieve elements and two viral forms of transport have been described: virions and ribonucleoprotein (RNP) complexes. With regard to poleroviruses, virions have always been presumed to be the long-distance transport form, but the potential role of RNP complexes has not been investigated. Here, we examined the requirement of virions for polerovirus systemic movement by analysing CP-targeted mutants that were unable to form viral particles. We confirmed that TuYV mutants that cannot encapsidate into virions are not able to reach systemic leaves. To completely discard the possibility that the introduced mutations in CP simply blocked the formation or the movement of RNP complexes, we tested in trans complementation of TuYV CP mutants by providing WT CP expressed in transgenic plants. WT CP was able to facilitate systemic movement of TuYV CP mutants and this observation was always correlated with the formation of virions. This demonstrated clearly that virus particles are essential for polerovirus systemic movement.

scholarly journals Capsid Protein Determinants Involved in Cell-to-Cell and Long Distance Movement of Tobacco Etch Potyvirus

Virology ◽  
1995 ◽  
Vol 206 (2) ◽  
pp. 1007-1016 ◽  
Author(s):  
Valerian V. Dolja ◽  
Ruth Haldeman-Cahill ◽  
Amanda E. Montgomery ◽  
Kathryn A. Vandenbosch ◽  
James C. Carrington
Keyword(s):  
Capsid Protein ◽  
Long Distance ◽  
Tobacco Etch Potyvirus ◽  
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 Systemic movement of a movement-deficient strain of Cucumber mosaic virus in zucchini squash is facilitated by a cucurbit-infecting potyvirus

2002 ◽  
Vol 83 (12) ◽  
pp. 3173-3178 ◽  
Author(s):  
Seung Kook Choi ◽  
Ju Yeon Yoon ◽  
Ki Hyun Ryu ◽  
Jang Kyung Choi ◽  
Peter Palukaitis ◽  
...  
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Cucurbita Pepo ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Long Distance ◽  
Systemic Movement ◽  
Tissue Print ◽  
Long Distance Movement

Zucchini squash (Cucurbita pepo) is a systemic host for most strains of the cucumovirus Cucumber mosaic virus (CMV), although the long-distance movement of the M strain of CMV (M-CMV) is inhibited in some cultivars. However, co-infection of zucchini plants with M-CMV and the potyvirus Zucchini yellow mosaic virus strain A (ZYMV-A) allowed M-CMV to move systemically, as demonstrated by tissue-print analysis. These doubly infected plants exhibited severe synergism in pathology. Infection of zucchini squash by M-CMV and an attenuated strain of ZYMV (ZYMV-AG) showed a milder synergy in pathology, in which ZYMV-AG also facilitated the long-distance movement of M-CMV similar to that promoted by ZYMV-A. Variation in the extent of synergy in pathology by the two strains of ZYMV did not correlate with differences in levels of accumulation of either virus. Thus, the extent of synergy in pathology is at least in part independent of the resistance-neutralizing function of the potyvirus.

scholarly journals Heterologous Replicase from Cucumoviruses can Replicate Viral RNAs, but is Defective in Transcribing Subgenomic RNA4A or Facilitating Viral Movement

Viruses ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 590 ◽  
Author(s):  
Shuangyu Gao ◽  
Jinda Lu ◽  
Xiaodong Cheng ◽  
Zhouhang Gu ◽  
Qiansheng Liao ◽  
...  
Keyword(s):  
Early Stage ◽  
Helicase Domain ◽  
Undetectable Level ◽  
Long Distance ◽  
Systemic Movement ◽  
Tomato Aspermy Virus ◽  
C Terminus ◽  
The Family ◽  
Viral Movement ◽  
Long Distance Movement

Interspecific exchange of RNA1 or RNA2 between the cucumoviruses cucumber mosaic virus (CMV) and tomato aspermy virus (TAV) was reported to be non-viable in plants previously. Here we investigated viability of the reassortants between CMV and TAV in Nicotiana benthamiana plants by Agrobacterium-mediated viral inoculation. The reassortants were composed of CMV RNA1 and TAV RNA2 plus RNA3 replicated in the inoculated leaves, while they were defective in viral systemic movement at the early stage of infection. Interestingly, the reassortant containing TAV RNA1 and CMV RNA2 and RNA3 infected plants systemically, but produced RNA4A (the RNA2 subgenome) at an undetectable level. The defect in production of RNA4A was due to the 1a protein encoded by TAV RNA1, and partially restored by replacing the C-terminus (helicase domain) in TAV 1a with that of CMV 1a. Collectively, exchange of the replicase components between CMV and TAV was acceptable for viral replication, but was defective in either directing transcription of subgenomic RNA4A or facilitating viral long-distance movement. Our finding may shed some light on evolution of subgenomic RNA4A in the family Bromoviridae.

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