Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation

Citrus leaf blotch virus has a single-stranded positive-sense genomic RNA (gRNA) of 8747 nt organized in three open reading frames (ORFs). The ORF1, encoding a polyprotein involved in replication, is translated directly from the gRNA, whereas ORFs encoding the movement (MP) and coat (CP) proteins ar...

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Main Authors: Renovell, Agueda, Gago-Zachert, Selma, Ruiz-Ruiz, Susana, Velázquez, Karelia, Navarro, Luis, Moreno, Pedro, Vives, María C., Guerri, José
Format: article
Language:Inglés
Published: 2017
Online Access:http://hdl.handle.net/20.500.11939/4409
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author Renovell, Agueda
Gago-Zachert, Selma
Ruiz-Ruiz, Susana
Velázquez, Karelia
Navarro, Luis
Moreno, Pedro
Vives, María C.
Guerri, José
author_browse Gago-Zachert, Selma
Guerri, José
Moreno, Pedro
Navarro, Luis
Renovell, Agueda
Ruiz-Ruiz, Susana
Velázquez, Karelia
Vives, María C.
author_facet Renovell, Agueda
Gago-Zachert, Selma
Ruiz-Ruiz, Susana
Velázquez, Karelia
Navarro, Luis
Moreno, Pedro
Vives, María C.
Guerri, José
author_sort Renovell, Agueda
collection ReDivia
description Citrus leaf blotch virus has a single-stranded positive-sense genomic RNA (gRNA) of 8747 nt organized in three open reading frames (ORFs). The ORF1, encoding a polyprotein involved in replication, is translated directly from the gRNA, whereas ORFs encoding the movement (MP) and coat (CP) proteins are expressed via 3' coterminal subgenomic RNAs (sgRNAs). We characterized the minimal promoter region critical for the CP-sgRNA expression in infected cells by deletion analyses using Agrobacterium-mediated infection of Nicotiana benthamiana plants. The minimal CP-sgRNA promoter was mapped between nucleotides -67 and +50 nt around the transcription start site. Surprisingly, larger deletions in the region between the CP-sgRNA transcription start site and the CP translation initiation codon resulted in increased CP-sgRNA accumulation, suggesting that this sequence could modulate the CP-sgRNA transcription. Site-specific mutational analysis of the transcription start site revealed that the +1 guanylate and the +2 adenylate are important for CP-sgRNA synthesis. (C) 2010 Elsevier Inc. All rights reserved.
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spelling ReDivia44092025-04-25T14:43:04Z Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation Renovell, Agueda Gago-Zachert, Selma Ruiz-Ruiz, Susana Velázquez, Karelia Navarro, Luis Moreno, Pedro Vives, María C. Guerri, José Citrus leaf blotch virus has a single-stranded positive-sense genomic RNA (gRNA) of 8747 nt organized in three open reading frames (ORFs). The ORF1, encoding a polyprotein involved in replication, is translated directly from the gRNA, whereas ORFs encoding the movement (MP) and coat (CP) proteins are expressed via 3' coterminal subgenomic RNAs (sgRNAs). We characterized the minimal promoter region critical for the CP-sgRNA expression in infected cells by deletion analyses using Agrobacterium-mediated infection of Nicotiana benthamiana plants. The minimal CP-sgRNA promoter was mapped between nucleotides -67 and +50 nt around the transcription start site. Surprisingly, larger deletions in the region between the CP-sgRNA transcription start site and the CP translation initiation codon resulted in increased CP-sgRNA accumulation, suggesting that this sequence could modulate the CP-sgRNA transcription. Site-specific mutational analysis of the transcription start site revealed that the +1 guanylate and the +2 adenylate are important for CP-sgRNA synthesis. (C) 2010 Elsevier Inc. All rights reserved. 2017-06-01T10:10:03Z 2017-06-01T10:10:03Z 2010 OCT 25 article Renovell, Agueda, Gago, Selma, Ruiz-Ruiz, S., Velazquez, Karelia, Navarro, L., Moreno, P., C. Vives, Mari, Guerri, J. (2010). Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation. Virology, 406(2), 360-369. 0042-6822 http://hdl.handle.net/20.500.11939/4409 10.1016/j.virol.2010.07.034 en openAccess Impreso
spellingShingle Renovell, Agueda
Gago-Zachert, Selma
Ruiz-Ruiz, Susana
Velázquez, Karelia
Navarro, Luis
Moreno, Pedro
Vives, María C.
Guerri, José
Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation
title Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation
title_full Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation
title_fullStr Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation
title_full_unstemmed Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation
title_short Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation
title_sort mapping the subgenomic rna promoter of the citrus leaf blotch virus coat protein gene by agrobacterium mediated inoculation
url http://hdl.handle.net/20.500.11939/4409
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