Structural alteration of a BYDV-like translation element (BTE) that attenuates p35 expression in three mild Tobacco bushy top virus isolates

Wang, Deya; Chang, Yu; Liu, Shanshan; Wang, Guolu; Shi, Kaichuang; Li, Xiangdong; Yuan, Xuefeng

doi:10.1038/s41598-017-04598-5

Cited by 13 publications

(19 citation statements)

References 36 publications

(53 reference statements)

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“…It usually increases the accumulation of one of the doubly infected viruses in the plant by facilitating its replication ( Vance, 1991 ; Karyeija et al, 2000 ) or movement to more tissues ( Barker, 1987 ; Zhou et al, 2017 ), or causes more severe symptoms in hosts than those caused by each single infection ( Untiveros et al, 2007 ; Syller, 2012 ). Synergistic interactions between poleroviruses and umbraviruses have destructive effects on crop plants ( Naidu et al, 1998 ; Mo et al, 2002 ). In previous studies, we have described that the N. benthamiana plants that co-infiltrated with BrYV and PEMV 2 developed mild leaf curling and produced chlorotic spots at 14 dpi.…”

Section: Discussionmentioning

confidence: 99%

The Conserved Proline18 in the Polerovirus P3a Is Important for Brassica Yellows Virus Systemic Infection

Zhang

Zhao

et al. 2018

Front. Microbiol.

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ORF3a, a newly identified non-AUG-initiated ORF encoded by members of genera Polerovirus and Luteovirus, is required for long-distance movement in plants. However, the mechanism of action of P3a in viral systemic movement is still not clear. In this study, sequencing of a brassica yellows virus (BrYV) mutant defective in systemic infection revealed two-nucleotide variation at positions 3406 and 3467 in the genome. Subsequent nucleotide substitution analysis proved that only the non-synonymous substitution (C→U) at position 3406, resulting in P3aP18L, abolished the systemic infection of BrYV. Preliminary investigation showed that wild type BrYV was able to load into the petiole of the agroinfiltrated Nicotiana benthamiana leaves, whereas the mutant displayed very low efficiency. Further experiments revealed that the P3a and its mutant P3aP18L localized to the Golgi apparatus and near plasmodesmata, as well as the endoplasmic reticulum. Both P3a and P3aP18L were able to self-interact in vivo, however, the mutant P3aP18L seemed to form more stable dimer than wild type. More interestingly, we confirmed firstly that the ectopic expression of P3a of other poleroviruses and luteoviruses, as well as co-infection with Pea enation mosaic virus 2 (PEMV 2), restored the ability of systemic movement of BrYV P3a defective mutant, indicating that the P3a is functionally conserved in poleroviruses and luteoviruses and is redundant when BrYV co-infects with PEMV 2. These observations provide a novel insight into the conserved function of P3a and its underlying mechanism in the systemic infection.

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Section: Discussionmentioning

confidence: 99%

The Conserved Proline18 in the Polerovirus P3a Is Important for Brassica Yellows Virus Systemic Infection

Zhang

Zhao

et al. 2018

Front. Microbiol.

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“…All plasmids were constructed based on the firefly luciferase reporter construct pT7-F-3′-UTRssp vector (Wang et al, 2017) via PCR amplification, enzyme digestion and ligation. All plasmids were confirmed by DNA sequencing.…”

Section: Construction Of Plasmids and Preparation Of Dna Fragmentsmentioning

confidence: 99%

“…In vitro translation assays were performed as previously described (Wang et al, 2017). Briefly, the in vitro-synthesized RNA transcripts (3 pmol) from designated translation reporter constructs were used for a 25 µL translation reaction volume using wheat germ extracts (WGE; Promega) according to the manufacturer's instructions.…”

Section: In Vitro Translationmentioning

confidence: 99%

A novel type of IRES having variable numbers of eIF4E-binding site and synergistic effect in RNA2 of WYMV isolates

Guowei

XiangDong

et al. 2021

Preprint

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Some viral proteins were translated in cap-independent manner via internal ribosome entry site (IRES), which ever maintained conservative characteristic among different isolates of same species of virus. However, IRES activity presented 7-fold of variance in RNA2 of wheat yellow mosaic virus (WYMV) HC and LYJN isolates. Based on RNA structure probing and mutagenesis assay, the loosened middle stem of H1 and hepta-nucleotide top loop of H2 in LYJN isolate synergistically ensured the higher IRES activity than that in HC isolate. In addition, the conserved top loop of H1 ensured basic IRES activity in HC and LYJN isolates. RNA2 5′-UTR specifically interacted with the wheat eIF4E, which was accomplished by the top loop of H1 in HC isolate or the top loop of H1 and H2 in LYJN isolate. Different IRES activity of WYMV RNA2 was regulated by different numbers of eIF4E-binding site and their synergistic effect, which was accomplished by the proximity of H1 and H2 due to the flexibility of middle stem in H1. It is represented a novel evolution pattern of IRES.

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“…All plasmids were constructed based on the firefly luciferase reporter construct pT7-F-3'UTRssp vector [28] and confirmed by DNA sequencing. pFluc-WY-R1-5 U was constructed by inserting the 5'UTR (162 nt) of WYMV RNA1 (GenBank: AF067124) between the T7 promoter and firefly luciferase (Fluc) gene in the pT7-F-3'UTRssp vector using BamH I and Sma I; the in vitro transcript from this plasmid was designated F-WY-R1-5 U. pFluc-WY-R1-3 U and pFluc-WY-R1-3 U·15A, with or without 15 adenylates, respectively, were separately constructed by inserting the 3'UTR (258 nt) of WYMV RNA1 (GenBank: AF067124) downstream of the Fluc gene in the pT7-F-3'UTRssp vector using Nru I and Ssp I; the in vitro transcript from these plasmids were designated F-WY-R1-3 U and F-WY-R1-3 U·15A.…”

Section: Plasmids and Dna Fragmentsmentioning

confidence: 99%

“…In vitro translation assays were performed as previously described [28]. Briefly, 3 pmol of in vitro-synthesized RNA transcripts from designated translation reporter constructs were used for a 25 μl translation reaction using wheat germ extracts (WGE; Promega) according to the manufacturer's instructions.…”

Section: In Vitro Translation Assaysmentioning

confidence: 99%

Variable 3’polyadenylation of Wheat yellow mosaic virus and its novel effects on translation and replication

Geng

Chang

et al. 2019

Virol J

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Background Polyadenylation influences many aspects of mRNA as well as viral RNA. variable polyadenylation at the 3' end have been reported in RNA viruses. It is interesting to identify the characteristic and potential role of 3' polyadenylation of Wheat yellow mosaic virus (WYMV), which has been reported to contain two genomic RNAs with 3' poly(A) tails and caused severe disease on wheat in East Asia region. Methods 3' RACE was used to identify sequences of the 3′ end in WYMV RNAs from naturally infected wheat by WYMV. In vitro translation assay was performed to analyze effect of UTRs of WYMV with or without 3’polyadenylation on translation. In vitro replication mediated by WYMV NIb protein were performed to evaluate effect of variable polyadenylation on replication. Results Variable polyadenylation in WYMV RNAs was identified via 3′ RACE. WYMV RNAs in naturally infected wheat in China simultaneously present with regions of long, short, or no adenylation at the 3' ends. The effects of variable polyadenylation on translation and replication of WYMV RNAs were evaluated. 5’UTR and 3’UTR of WYMV RNA1 or RNA2 synergistically enhanced the translation of the firefly luciferase ( Fluc ) gene in in vitro WGE system, whereas additional adenylates had an oppositive effect on this enhancement on translation mediated by UTRs of WYMV. Additional adenylates remarkably inhibited the synthesis of complementary strand from viral genome RNA during the in vitro replication mediated by WYMV NIb protein. Conclusions 3' end of WYMV RNAs present variable polyadenylation even no polyadenylation. 3' polyadenylation have opposite effect on translation mediated by UTRs of WYMV RNA1 or RNA2. 3' polyadenylation have negative effect on minus-strand synthesis of WYMV RNA in vitro. Variable polyadenylation of WYMV RNAs may provide sufficient selection on the template for translation and replication. Electronic supplementary material The online version of this article (10.1186/s12985-019-1130-z) contains supplementary material, which is available to authorized users.

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Structural alteration of a BYDV-like translation element (BTE) that attenuates p35 expression in three mild Tobacco bushy top virus isolates

Cited by 13 publications

References 36 publications

The Conserved Proline18 in the Polerovirus P3a Is Important for Brassica Yellows Virus Systemic Infection

The Conserved Proline18 in the Polerovirus P3a Is Important for Brassica Yellows Virus Systemic Infection

A novel type of IRES having variable numbers of eIF4E-binding site and synergistic effect in RNA2 of WYMV isolates

Variable 3’polyadenylation of Wheat yellow mosaic virus and its novel effects on translation and replication

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