Properties of African Cassava Mosaic Virus Capsid Protein Expressed in Fission Yeast

Hipp, Katharina; Schafer, Benjamin W.; Kepp, Gabi; Jeske, Holger

doi:10.3390/v8070190

Cited by 3 publications

(11 citation statements)

References 67 publications

(103 reference statements)

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“…Geminivirus proteins, including the begomovirus NSP, movement protein, AC3, AC4, BC1, BV1 and capsid protein, the curtovirus C4 and AL2/C2 proteins, and the satellite ␤C1 protein, are also substrates of host protein kinases during plant-virus interactions (27,28,(44)(45)(46)(47)(48)(49). It is well established that host Ser/Thr phosphorylation of plant viral proteins plays vital roles in attenuation of symptom development and efficiency of viral infection (28,(48)(49)(50)(51)(52).…”

Section: Discussionmentioning

confidence: 99%

Mimic Phosphorylation of a βC1 Protein Encoded by TYLCCNB Impairs Its Functions as a Viral Suppressor of RNA Silencing and a Symptom Determinant

Zhong

Wang

Xiao

et al. 2017

J Virol

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Phosphorylation of the ␤C1 protein encoded by the betasatellite of tomato yellow leaf curl China virus (TYLCCNB-␤C1) by SNF1-related protein kinase 1 (SnRK1) plays a critical role in defense of host plants against geminivirus infection in Nicotiana benthamiana. However, how phosphorylation of TYLCCNB-␤C1 impacts its pathogenic functions during viral infection remains elusive. In this study, we identified two additional tyrosine residues in TYLCCNB-␤C1 that are phosphorylated by SnRK1. The effects of TYLCCNB-␤C1 phosphorylation on its functions as a viral suppressor of RNA silencing (VSR) and a symptom determinant were investigated via phosphorylation mimic mutants in N. benthamiana plants. Mutations that mimic phosphorylation of TYLCCNB-␤C1 at tyrosine 5 and tyrosine 110 attenuated disease symptoms during viral infection. The phosphorylation mimics weakened the ability of TYLCCNB-␤C1 to reverse transcriptional gene silencing and to suppress posttranscriptional gene silencing and abolished its interaction with N. benthamiana ASYMMETRIC LEAVES 1 in N. benthamiana leaves. The mimic phosphorylation of TYLCCNB-␤C1 had no impact on its protein stability, subcellular localization, or self-association. Our data establish an inhibitory effect of phosphorylation of TYLCCNB-␤C1 on its pathogenic functions as a VSR and a symptom determinant and provide a mechanistic explanation of how SnRK1 functions as a host defense factor. IMPORTANCE Tomato yellow leaf curl China virus (TYLCCNV), which causes a severe yellow leaf curl disease in China, is a monopartite geminivirus associated with the betasatellite (TYLCCNB). TYLCCNB encodes a single pathogenicity protein, ␤C1 (TYLCCNB-␤C1), which functions as both a viral suppressor of RNA silencing (VSR) and a symptom determinant. Here, we show that mimicking phosphorylation of TYLCCNB-␤C1 weakens its ability to reverse transcriptional gene silencing, to suppress posttranscriptional gene silencing, and to interact with N. benthamiana ASYMMETRIC LEAVES 1. To our knowledge, this is the first report establishing an inhibitory effect of phosphorylation of TYLCCNB-␤C1 on its pathogenic functions as both a VSR and a symptom determinant and to provide a mechanistic explanation of how SNF1-related protein kinase 1 acts as a host defense factor. These findings expand the scope of phosphorylation-mediated defense mechanisms and contribute to further understanding of plant defense mechanisms against geminiviruses.KEYWORDS TYLCCNB-␤C1, Nicotiana benthamiana, SnRK1, geminivirus, host defense factor, posttranscriptional gene silencing, protein phosphorylation, transcriptional gene silencing

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Section: Discussionmentioning

confidence: 99%

Mimic Phosphorylation of a βC1 Protein Encoded by TYLCCNB Impairs Its Functions as a Viral Suppressor of RNA Silencing and a Symptom Determinant

Zhong

Wang

Xiao

et al. 2017

J Virol

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“…Fig. 2, and Hipp et al (2016)). The calculated apparent molecular masses were determined as 29-30 kDa for pCP and the lower yCP band, whereas 31-32 kDa were determined for the upper yCP band in different gels (Fig.…”

Section: Only a Single Cp Band Was Detected In Total Proteins From Plantsmentioning

confidence: 90%

“…The correctness of the constructs was confirmed by sequencing (Macrogene Europe, Amsterdam, The Netherlands). Compared to the database entry (GenBank accession number AJ427910), pUC19-APA9 showed a nucleotide substitution from CT to AG at position 164/165 (nucleotide numbering according to the AV1 sequence) resulting in an amino acid substitution T55M (Hipp et al, 2016).…”

Section: Construction Of Plasmidsmentioning

confidence: 99%

“…Here, other protein-protein or protein-nucleic acid interactions may be needed for the production of mature geminivirions (Hesketh et al, 2018;Hipp et al, 2017). Two CP bands were found after SDS gel electrophoresis, depending on the virus species and the host (Abouzid and Jeske, 1986;Gorovits et al, 2013aGorovits et al, , 2013b as well as after ectopic expression in fission yeast (referred to as yeast) (Hipp et al, 2016). The different migration behavior may be indicative for post-translational modification of CP, which has been investigated now in plants, in total protein extracts and mature virions.…”

Section: Introductionmentioning

confidence: 99%

“…First indications for a post-translational modification were found for Abutilon mosaic virus (AbMV) CP expressed and phosphorylated in Escherichia coli (Wege and Jeske, 1998). The precise positions of phosphorylated amino acids in the sequence of the ACMV CP (T12, S25 and S62) were determined by mass spectrometry (MS) (Hipp et al, 2016) after expression of the CP gene in yeast (yCP). The majority of yCP migrated more slowly in SDS gels than ACMV CP from purified particles (pvCP).…”

Section: Introductionmentioning

confidence: 99%

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Different forms of African cassava mosaic virus capsid protein within plants and virions

et al. 2019

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Identification of the Viral Determinant of Hypovirulence and Host Range in Sclerotiniaceae of a Genomovirus Reconstructed from the Plant Metagenome

Feng

Wang

et al. 2021

J Virol

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Uncharacterized viral genomes that encode circular replication-associated proteins of single-stranded DNA viruses have been discovered by metagenomics/metatranscriptomics approaches. Some of these novel viruses are classified under the newly formed Genomoviridae family. Here, we determine the host range of a novel genomovirus, SlaGemV-1, through the transfection of Sclerotinia sclerotiorum with infectious clones. Inoculating with the rescued virions, we further transfected Botrytis cinerea and Monilinia fruticola , two economically important members of family Sclerotiniaceae, and Fusarium oxysporum . SlaGemV-1 causes hypovirulence in S. sclerotiorum, B. cinerea , and M. fruticola . SlaGemV-1 also replicates in Spodoptera frugiperda insect cells, but not in Caenorhabditis elegans nor plants. By expressing viral-encoded genes separately through site-specific integration, the replication protein alone was sufficient to cause debilitation. Our study is the first to demonstrate the reconstruction of a metagenomically discovered genomovirus without known hosts with the potential of inducing hypovirulence, and the infectious clone allows for studying mechanisms of genomovirus-host interactions that are conserved across genera. Importance Little is known about the exact host range of widespread genomoviruses. The genome of soybean leaf-associated gemygorvirus-1 (SlaGemV-1) was originally assembled from a metagenomic/metatranscriptomic study without known hosts. Here, we rescued SlaGemV-1 and found that it could infect three important plant pathogenic fungi and Fall armyworm (S. frugiperda , Sf9) insect cells, but not a model nematode, C. elegans , or model plant species. Most importantly, SlaGemV-1 shows promise for inducing hypovirulence of the tested fungal species under family Sclerotiniaceae, including Sclerotinia sclerotiorum , Botrytis cinerea , Monilinia fruticola . The viral determinant of hypovirulence was further identified as replication initiation protein. As a proof of concept, we demonstrate that viromes discovered in plant metagenome can be a valuable genetic resource when novel viruses are rescued and characterized for their host range.

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Properties of African Cassava Mosaic Virus Capsid Protein Expressed in Fission Yeast

Cited by 3 publications

References 67 publications

Mimic Phosphorylation of a βC1 Protein Encoded by TYLCCNB Impairs Its Functions as a Viral Suppressor of RNA Silencing and a Symptom Determinant

Mimic Phosphorylation of a βC1 Protein Encoded by TYLCCNB Impairs Its Functions as a Viral Suppressor of RNA Silencing and a Symptom Determinant

Different forms of African cassava mosaic virus capsid protein within plants and virions

Identification of the Viral Determinant of Hypovirulence and Host Range in Sclerotiniaceae of a Genomovirus Reconstructed from the Plant Metagenome

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