Spontaneous Mutation in the Movement Protein of Citrus Leprosis Virus C2, in a Heterologous Virus Infection Context, Increases Cell-to-Cell Transport and Generates Fitness Advantage

Previous results using a movement defective alfalfa mosaic virus (AMV) vector revealed that citrus leprosis virus C (CiLV-C) movement protein (MP) generates a more efficient local movement, but not more systemic transport, than citrus leprosis virus C2 (CiLV-C2) MP, MPs belonging to two important viruses for the citrus industry. Here, competition experiment assays in transgenic tobacco plants (P12) between transcripts of AMV constructs expressing the cilevirus MPs, followed by several biological passages, showed the prevalence of the AMV construct carrying the CiLV-C2 MP. The analysis of AMV RNA 3 progeny recovered from P12 plant at the second viral passage revealed the presence of a mix of progeny encompassing the CiLV-C2 MP wild type (MPWT) and two variants carrying serines instead phenylalanines at positions 72 (MPS72F) or 259 (MPS259F), respectively. We evaluated the effects of each modified residue in virus replication, and cell-to-cell and long-distance movements. Results indicated that phenylalanine at position 259 favors viral cell-to-cell transport with an improvement in viral fitness, but has no effect on viral replication, whereas mutation at position 72 (MPS72F) has a penalty in the viral fitness. Our findings indicate that the prevalence of a viral population may be correlated with its greater efficiency in cell-to-cell and systemic movements.

[1]  E. Kitajima,et al.  Unravelling the involvement of cilevirus p32 protein in the viral transport , 2021, Scientific reports.

[2]  E. Kitajima,et al.  Membrane Association and Topology of Citrus Leprosis Virus C2 Movement and Capsid Proteins , 2021, Microorganisms.

[3]  Y. Dorokhov,et al.  Diversity of Plant Virus Movement Proteins: What Do They Have in Common? , 2020, Processes.

[4]  E. Kitajima,et al.  Dichorhaviruses Movement Protein and Nucleoprotein Form a Protein Complex That May Be Required for Virus Spread and Interacts in vivo With Viral Movement-Related Cilevirus Proteins , 2020, Frontiers in Microbiology.

[5]  E. Kitajima,et al.  Citrus Leprosis Virus C Encodes Three Proteins With Gene Silencing Suppression Activity , 2020, Frontiers in Microbiology.

[6]  E. Kitajima,et al.  Brevipalpus-transmitted viruses: parallelism beyond a common vector or convergent evolution of distantly related pathogens? , 2018, Current opinion in virology.

[7]  Marilia S. Silva,et al.  Dissecting the Subcellular Localization, Intracellular Trafficking, Interactions, Membrane Association, and Topology of Citrus Leprosis Virus C Proteins , 2018, Front. Plant Sci..

[8]  S. Li,et al.  Single amino acid in V2 encoded by TYLCV is responsible for its self-interaction, aggregates and pathogenicity , 2018, Scientific Reports.

[9]  V. Pallás,et al.  The functional analysis of distinct tospovirus movement proteins (NSM) reveals different capabilities in tubule formation, cell-to-cell and systemic virus movement among the tospovirus species. , 2017, Virus research.

[10]  C. T. Anderson,et al.  Identification of Ourmiavirus 30K movement protein amino acid residues involved in symptomatology, viral movement, subcellular localization and tubule formation. , 2016, Molecular plant pathology.

[11]  A. Obrępalska-Stęplowska,et al.  A single amino acid substitution in movement protein of tomato torrado virus influences ToTV infectivity in Solanum lycopersicum. , 2016, Virus research.

[12]  M. Melzer,et al.  Role Bending: Complex Relationships Between Viruses, Hosts, and Vectors Related to Citrus Leprosis, an Emerging Disease. , 2015, Phytopathology.

[13]  J. García,et al.  Viral factors involved in plant pathogenesis. , 2015, Current opinion in virology.

[14]  S. Elena,et al.  Evolution of plant virus movement proteins from the 30K superfamily and of their homologs integrated in plant genomes. , 2015, Virology.

[15]  E. Moriones,et al.  The movement protein (NSm) of Tomato spotted wilt virus is the avirulence determinant in the tomato Sw-5 gene-based resistance. , 2014, Molecular plant pathology.

[16]  R.-H. Wen,et al.  Fitness penalty in susceptible host is associated with virulence of Soybean mosaic virus on Rsv1-genotype soybean: a consequence of perturbation of HC-Pro and not P3. , 2013, Molecular plant pathology.

[17]  R. Brlansky,et al.  A novel virus of the genus Cilevirus causing symptoms similar to citrus leprosis. , 2013, Phytopathology.

[18]  V. Pallás,et al.  Systemic transport of Alfalfa mosaic virus can be mediated by the movement proteins of several viruses assigned to five genera of the 30K family. , 2013, The Journal of general virology.

[19]  J. García,et al.  How do plant viruses induce disease? Interactions and interference with host components. , 2011, The Journal of general virology.

[20]  E. Massad,et al.  Modeling the competition between viruses in a complex plant-pathogen system. , 2010, Phytopathology.

[21]  V. Pallás,et al.  Implication of the C terminus of the Prunus necrotic ringspot virus movement protein in cell-to-cell transport and in its interaction with the coat protein. , 2010, The Journal of general virology.

[22]  E. Kitajima,et al.  Citrus Leprosis: Centennial of an Unusual Mite-Virus Pathosystem. , 2010, Plant disease.

[23]  V. Pallás,et al.  Caulimoviridae Tubule-Guided Transport Is Dictated by Movement Protein Properties , 2010, Journal of Virology.

[24]  E. Massad,et al.  Modeling the Dynamics of Viral Evolution Considering Competition Within Individual Hosts and at Population Level: The Effects of Treatment , 2010, Bulletin of mathematical biology.

[25]  D. J. Lewandowski,et al.  Identification of domains of the Tomato spotted wilt virus NSm protein involved in tubule formation, movement and symptomatology. , 2009, Virology.

[26]  E. Kitajima,et al.  Natural Infection of Swinglea glutinosa by the Citrus leprosis virus Cytoplasmic Type (CiLV-C) in Colombia. , 2008, Plant disease.

[27]  A. A. Souza,et al.  Complete nucleotide sequence, genomic organization and phylogenetic analysis of Citrus leprosis virus cytoplasmic type. , 2006, The Journal of general virology.

[28]  P. Arruda,et al.  The Complete Nucleotide Sequence and Genomic Organization of Citrus Leprosis Associated Virus, Cytoplasmatic type (CiLV-C) , 2006, Virus Genes.

[29]  V. Pallás,et al.  Cell-to-cell movement of Alfalfa mosaic virus can be mediated by the movement proteins of Ilar-, bromo-, cucumo-, tobamo- and comoviruses and does not require virion formation. , 2006, Virology.

[30]  W. J. Lucas,et al.  Plant viral movement proteins: agents for cell-to-cell trafficking of viral genomes. , 2006, Virology.

[31]  D. J. Lewandowski,et al.  The tubule-forming NSm protein from Tomato spotted wilt virus complements cell-to-cell and long-distance movement of Tobacco mosaic virus hybrids. , 2005, Virology.

[32]  P. Palukaitis,et al.  Cucumber mosaic virus 2a polymerase and 3a movement proteins independently affect both virus movement and the timing of symptom development in zucchini squash. , 2005, The Journal of general virology.

[33]  J. Bol,et al.  Role of the alfalfa mosaic virus movement protein and coat protein in virus transport. , 2001, Molecular plant-microbe interactions : MPMI.

[34]  D T Jones,et al.  Protein secondary structure prediction based on position-specific scoring matrices. , 1999, Journal of molecular biology.

[35]  B. Ding.  Intercellular protein trafficking through plasmodesmata , 1998, Plant Molecular Biology.

[36]  W. J. Lucas,et al.  Mutations in viral movement protein alter systemic infection and identify an intercellular barrier to entry into the phloem long-distance transport system. , 1998, Virology.

[37]  I. Furusawa,et al.  Deletion of the C-terminal 33 amino acids of cucumber mosaic virus movement protein enables a chimeric brome mosaic virus to move from cell to cell , 1997, Journal of virology.

[38]  Alison G. Power,et al.  Competition between Viruses in a Complex Plant‐‐Pathogen System , 1996 .

[39]  V. Pallás,et al.  Non-isotopic tissue-printing hybridization: a new technique to study long-distance plant virus movement. , 1995, Journal of virological methods.

[40]  P. Taschner,et al.  Replication of an incomplete alfalfa mosaic virus genome in plants transformed with viral replicase genes. , 1991, Virology.

[41]  W. J. Lucas,et al.  Movement Protein of Tobacco Mosaic Virus Modifies Plasmodesmatal Size Exclusion Limit , 1989, Science.

[42]  J. Bol,et al.  Expression of alfalfa mosaic virus cDNA1 and 2 in transgenic tobacco plants. , 1988, Virology.

[43]  T. Hall,et al.  Expression of alfalfa mosaic virus RNA 4 cDNA transcripts in vitro and in vivo. , 1985, Virology.

[44]  J. Navarro,et al.  Key checkpoints in the movement of plant viruses through the host. , 2019, Advances in virus research.

[45]  Nick V Grishin,et al.  PROMALS3D: multiple protein sequence alignment enhanced with evolutionary and three-dimensional structural information. , 2014, Methods in molecular biology.

[46]  M. Heinlein,et al.  Cellular pathways for viral transport through plasmodesmata , 2010, Protoplasma.

[47]  L. Torrance,et al.  Role of plant virus movement proteins. , 2008, Methods in molecular biology.

[48]  M. Heinlein,et al.  Macromolecular transport and signaling through plasmodesmata. , 2004, International review of cytology.

[49]  M. Suzuki,et al.  Combination of amino acids in the 3a protein and the coat protein of Cucumber mosaic virus determines symptom expression and viral spread in bottle gourd , 2001, Archives of Virology.

[50]  J. Bol,et al.  Engineering of Alfalfa mosaic virus RNA 3 into an expression vector , 2001, Archives of Virology.

[51]  U. Melcher.  The '30K' superfamily of viral movement proteins. , 2000, The Journal of general virology.

[52]  V. Pallás,et al.  Detection of plant RNA viruses by nonisotopic dot-blot hybridization. , 1998, Methods in molecular biology.

[53]  D. Klessig,et al.  A single amino acid change in turnip crinkle virus movement protein p8 affects RNA binding and virulence on Arabidopsis thaliana. , 1998, Journal of virology.