Insights into RNA Virus Mutant Spectrum and Lethal Mutagenesis Events: Replicative Interference and Complementation by Multiple Point Mutants

Perales, Celia; Mateo, Roberto; Mateu, Mauricio G.; Domingo, Esteban

doi:10.1016/j.jmb.2007.03.074

Cited by 91 publications

(116 citation statements)

References 51 publications

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“…However, no attempt was made, to our knowledge, to quantify the efficiency of complementation of DIP by helper viruses. Quantitative analyses of complementation have been reported for animal viruses in cell culture (e.g., Novella et al, 2004;Wilke et al, 2004;Perales et al, 2007), but dynamics of infection in cell culture an in the multicellular animal or plant host could vary largely. The few reported quantitative analyses of complementation in the multicellular host come all from the plant virus field.…”

Section: Discussionmentioning

confidence: 99%

A quantitative analysis of complementation of deleterious mutants in plant virus populations

Fraile

Sacristán

García‐Arenal

2008

Span. j. agric. res.

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Complementation can be defined as the process by which the function affected by a mutation is provided in trans by fully competent genotypes. Complementation can, thus, counter the effects of selection of deleterious mutants. Complementation of mutants defective for replication, movement and transmission has been often described in experiments with viruses and, occasionally, has been reported to occur in their natural populations. However, the role of complementation in virus evolution has been overlooked. Here is provided a quantitative estimate of the efficiency of complementation, defined as the probability that a non-functional mutant accomplishes a function relative to a functional one. For this, the frequency of mutants of Tobacco mosaic virus (TMV) defective for cell to cell movement was estimated in wild type tobacco plants and in transgenic plants expressing the TMV movement protein (MP) from a transgene. Mutants lethal for cell-to-cell movement were complemented by wild-type TMV in the first case, and by the transgene-expressed MP in the second case. Assuming that complementation is fully efficient in the transgenic plants, a value for the efficiency of complementation of 0.34 was obtained. Thus, complementation can efficiently counter selection on lethal mutants, and may have an important role on virus evolution. Complementation may be relevant for management of viral diseases if the complemented deleterious mutation is linked to other functions affecting the pathogenicity or epidemiology of the virus.Additional key words: mutant complementation, selection, Tobacco mosaic virus, virus evolution. Resumen Análisis cuantitativo de la complementación de mutantes deletéreos en poblaciones de virus vegetalesLa complementación puede definirse como el proceso por el cual una función afectada por una mutación es provista en trans por genotipos competentes para dicha función. La complementación de mutantes defectivos para replicación, movimiento o transmisión se ha descrito con frecuencia en poblaciones experimentales de virus y, ocasionalmente, en poblaciones de campo. Sin embargo, el papel de la complementación en la evolución de los virus no se ha considerado. En este trabajo presentamos una estima de la eficacia de la complementación, definida como la probabilidad de que un mutante no funcional realice la función respecto a un genotipo funcional. Para ello se estimó la frecuencia de mutantes del virus del mosaico del tabaco (TMV) defectivos para el movimiento de célula a célula en plantas silvestres de tabaco y en plantas transgénicas que expresan constitutivamente la proteína del movimiento (MP) de TMV. En el primer caso, los mutantes letales para movimiento serán complementados por genotipos silvestres de TMV, y en el segundo, por la MP transgénica. Suponiendo que la complementación en las plantas transgénicas es totalmente eficaz, se obtuvo un valor de la eficacia de complementación de 0,34. Esto muestra que la complementación puede contrarrestar eficazmente el efecto de la selección sobre mutant...

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Section: Discussionmentioning

confidence: 99%

A quantitative analysis of complementation of deleterious mutants in plant virus populations

Fraile

Sacristán

García‐Arenal

2008

Span. j. agric. res.

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“…In other virus systems, specific interference by viral mutants required that the interfering genomes be replication-competent (Crowder & Kirkegaard, 2005;Perales et al, 2007). To test whether interfering preparations expressed viral antigens, the supernatants of cultures persistently infected with LCMV in the presence of FU, either with or without detectable infectivity [culture media obtained at 96 h post-infection (p.i.)…”

Section: Detection Of Virus-specific Antigens and Virus Particles In mentioning

confidence: 99%

“…Parallel studies carried out with the picornavirus foot-and-mouth disease virus (FMDV) have provided independent evidence for the suppressive effects of mutant spectra (González-Ló pez et al, 2004). In particular, well-characterized capsid and polymerase mutants of FMDV exerted a specific interference on FMDV RNA replication, and contributed to extinction (Perales et al, 2007(Perales et al, , 2009). …”

Section: Introductionmentioning

confidence: 99%

An interfering activity against lymphocytic choriomeningitis virus replication associated with enhanced mutagenesis

Martı́n

Abia

Domingo

et al. 2009

Journal of General Virology

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Previous studies have documented that, in the presence of the mutagenic base analogue 5-fluorouracil (FU), lymphocytic choriomeningitis virus (LCMV) that persisted in BHK-21 cells decreased its infectivity to a larger extent than intracellular viral RNA levels, prior to virus extinction. This observation, together with in silico simulations, led to the proposal of the lethal defection model of virus extinction. This model suggests the participation of defective-interfering genomes in the loss of infectivity by increased mutagenesis. Since LCMV naturally produces defective-interfering particles, it was important to show that a capacity to interfere is produced in association with FU treatment. Here, we document that BHK-21 cells persistently infected with LCMV grown in the presence of FU, but not in its absence, generated an interfering activity that suppressed LCMV infectivity. Interference was specific for LCMV and was sensitive to UV irradiation and its activity was dose-and time-dependent. The interfering preparations produced positive LCMV immunofluorescence and viral particles seen by electron microscopy when used to infect cells, despite some preparations being devoid of detectable infectivity. Interference did not involve significant increases of mutant spectrum complexity, as predicted by the lethal defection model. The results provide support for a specific interference associated with LCMV when the virus replicates in the presence of FU. The excess of interference relative to that observed in the absence of FU is necessary for LCMV extinction.

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“…The amount of extract analysed was normalized using actin, identified with a specific mAb (anti-b-actin clone AC-15; Sigma). Actin was chosen due to its long intracellular half-life, and the amount of extract analysed corresponded to the linear region of the relationship between the amount of extract and the intensity of the actin band by Western blot (Perales et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

“…Then, an adequate dilution in PBS (with 0.1 % powdered milk) of the primary antibody was added, and the membrane incubated for 2 h. The membrane was then washed three times for 15 min in TPBS (Tween 20 at 0.05 %,v/v,in PBS) and incubated with the corresponding secondary antibody conjugated to peroxidase (1 : 10 000 dilution in TPBS). After 1 h of incubation, three washes with PBS were carried out, and the membrane was developed by chemiluminescence (ECL; Amersham) (Herrera et al, 2008;Moreno et al, 2014;Perales et al, 2007). The amount of cell extract used for electrophoretic analysis was normalized to a constant amount of cellular actin, measured by reactivity with monoclonal antibody (anti-b-actin clone AC-15; Sigma), and corresponded to a concentration of protein in the linear region of the relationship between the Western blot signal and the protein concentration.…”

Section: Methodsmentioning

confidence: 99%

Distance effects during polyprotein processing in the complementation between defective FMDV RNAs

Moreno

Perales

2016

Journal of General Virology

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Passage of foot-and-mouth disease virus (FMDV) in BHK-21 cells resulted in the segmentation of the viral genome into two defective RNAs lacking part of either the L-or the capsid-coding region. The two RNAs are infectious by complementation. Electroporation of L-defective RNA in BHK-21 cells resulted in the accumulation of the precursor P3 located away from the deleted sequence. Expression of L in trans led to the processing of P3, indicating that there is a connection between L protease activity and the secondary cleavages carried out by 3C protease within P3. These results suggest that the complementation mechanism between defective RNAs is not restricted to supplying the L and capsid proteins but that distance effects on polyprotein processing events are also implicated.

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Insights into RNA Virus Mutant Spectrum and Lethal Mutagenesis Events: Replicative Interference and Complementation by Multiple Point Mutants

Cited by 91 publications

References 51 publications

A quantitative analysis of complementation of deleterious mutants in plant virus populations

A quantitative analysis of complementation of deleterious mutants in plant virus populations

An interfering activity against lymphocytic choriomeningitis virus replication associated with enhanced mutagenesis

Distance effects during polyprotein processing in the complementation between defective FMDV RNAs

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