Mitogen‐activated protein kinase phosphatase 1 reduces the replication efficiency of <i>Bamboo mosaic virus</i> in <i>Nicotiana benthamiana</i>

Lee, Cheng-Cheng; Wu, Yan; Hsueh, Chia‐Hsin; Huang, Yu‐Ting; Hsu, Yau‐Heiu; Meng, Menghsiao

doi:10.1111/mpp.12701

Cited by 7 publications

(8 citation statements)

References 51 publications

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“…Plasmids. Binary plasmids pERep-HA and pKSF4 were used to produce the hemagglutinin tag-fused REP BaMV (REP BaMV -HA) and the satBaMV SF4 variant, respectively, in N. benthamiana as in previous reports (9,10). The cDNA encoding N. benthamiana PCNA was cloned by PCR from a leaf cDNA preparation according to the transcriptomic information available in the N. benthamiana Genome and Transcriptome Sequencing Consortium website (http://benthgenome.qut.edu.au/).…”

Section: Methodsmentioning

confidence: 99%

“…An infectious clone pCBG, containing a duplicated CP promoter-driven GFP gene, enabled us to monitor the replication and systemic movement of BaMV in host plants based on the expression of GFP (3). In this study, pCBG and pCF were used to establish the infection of BaMV and Foxtail mosaic virus (FoMV) in protoplasts, respectively (4,10). pXTAL, an infectious clone of Potato virus X (PVX), was constructed by substituting the BaMV cDNA in BaMV infectious clone with the cDNA prepared from a local PVX strain (GenBank accession no.…”

Section: Methodsmentioning

confidence: 99%

“…Foxtail mosaic virus (FoMV), phylogenetically close to BaMV, shares several similar host-pathogen interactions to BaMV when they proliferate in N. benthamiana. For example, the plant MAPK phosphatase 1, a negative regulator in the MAPK pathway, reduces the replication efficiency of both BaMV and FoMV (10), and the cytoplasmic 5=¡3= exonuclease increases the accumulation level of BaMV as well as FoMV (9). The effect of PCNA on FoMV was also tested in this study.…”

Section: Figmentioning

confidence: 99%

“…Moreover, SF4 may facilitate the recruitment of host factors to constitute a competent viral replication complex. In fact, the REP BaMV -containing membrane fraction prepared from the agroinfiltrated N. benthamiana has facilitated the in vitro RdRp activity assay by using the endogenous SF4 as the template (9,10). It is worth noting that addition of ionic detergents, e.g., 0.1% sodium dodecyl sulfate (SDS), in the REP BaMV -containing membrane preparation was able to boost the in vitro BaMV RdRp activity, suggesting a strong physical stability of the membrane-associated viral replication complex.…”

mentioning

confidence: 99%

“…Among them, a cytoplasmic 5=¡3= exoribonuclease is capable of increasing BaMV accumulation by presumably removing uncapped aberrant viral RNAs, which otherwise might interfere with the replication of normal viral RNAs (9). The N. benthamiana mitogen-activated protein kinase phosphatase 1 (NbMKP1), a negative regulator in the plant mitogen-activated protein kinase (MAPK) pathway, attenuates the replication efficiency of BaMV, suggesting that an environment favorable for BaMV replication is created by the activation of MAPK pathway (10).…”

mentioning

confidence: 99%

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Proliferating Cell Nuclear Antigen Suppresses RNA Replication of Bamboo Mosaic Virus through an Interaction with the Viral Genome

Lee

Wang

Leu

et al. 2019

J Virol

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Bamboo mosaic virus (BaMV), a member of the Potexvirus genus, has a monopartite positive-strand RNA genome on which five open reading frames (ORFs) are organized. ORF1 encodes a 155-kDa nonstructural protein (REPBaMV) that plays a core function in replication/transcription of the viral genome. To find out cellular factors modulating the replication efficiency of BaMV, a putative REPBaMV-associated protein complex from Nicotiana benthamiana leaf was isolated on an SDS-PAGE gel, and a few proteins preferentially associated with REPBaMV were identified by tandem mass spectrometry. Among them, proliferating cell nuclear antigen (PCNA) was particularly noted. Overexpression of PCNA strongly suppressed the accumulation of BaMV coat protein and RNAs in leaf protoplasts. In addition, PCNA exhibited an inhibitory effect on BaMV polymerase activity. A pulldown assay confirmed a binding capability of PCNA toward BaMV genomic RNA. Mutations at D41 or F114 residues, which are critical for PCNA to function in nuclear DNA replication and repair, disabled PCNA from binding BaMV genomic RNA as well as suppressing BaMV replication. This suggests that PCNA bound to the viral RNA may interfere with the formation of a potent replication complex or block the replication process. Interestingly, BaMV is almost invisible in the newly emerging leaves where PCNA is actively expressed. Accordingly, PCNA is probably one of the factors restricting the proliferation of BaMV in young leaves. Foxtail mosaic virus and Potato virus X were also suppressed by PCNA in the protoplast experiment, suggesting a general inhibitory effect of PCNA on the replication of potexviruses. IMPORTANCE Knowing the dynamic interplay between plant RNA viruses and their host is a basic step toward first understanding how the viruses survive the plant defense mechanisms and second gaining knowledge of pathogenic control in the field. This study found that plant proliferating cell nuclear antigen (PCNA) imposes a strong inhibition on the replication of several potexviruses, including Bamboo mosaic virus, Foxtail mosaic virus, and Potato virus X. Based on the tests on Bamboo mosaic virus, PCNA is able to bind the viral genomic RNA, and this binding is a prerequisite for the protein to suppress the virus replication. This study also suggests that PCNA plays an important role in restricting the proliferation of potexviruses in the rapidly dividing tissues of plants.

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Section: Methodsmentioning

confidence: 99%