The interaction of Lolium latent virus major coat protein with ankyrin repeat protein NbANKr redirects it to chloroplasts and modulates virus infection

The Lolium latent virus (LoLV) major coat protein sequence contains a typical chloroplast transit peptide (cTP) domain. In infected Nicotiana benthamiana leaf tissue, LoLV coat proteins can be detected at the chloroplast. In transient expression, several N-terminal deletions of the CP sequence, incr...

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Bibliographic Details
Published in:Journal of general virology 2018-05, Vol.99 (5), p.730-742
Main Authors: Vaira, A M, Lim, H S, Bauchan, G, Gulbronson, C J, Miozzi, L, Vinals, N, Natilla, A, Hammond, J
Format: Article
Language:English
Subjects:
Ankyrin Repeat
Capsid Proteins - genetics
Chloroplast Proteins - genetics
Chloroplasts - virology
Gene Expression Regulation, Plant
Gene Silencing
Nicotiana - virology
Plant Diseases - virology
Plant Leaves - virology
Plant Proteins - genetics
Plant Viruses - genetics
Protein Sorting Signals - genetics
RNA, Viral - genetics
Two-Hybrid System Techniques
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Summary:The Lolium latent virus (LoLV) major coat protein sequence contains a typical chloroplast transit peptide (cTP) domain. In infected Nicotiana benthamiana leaf tissue, LoLV coat proteins can be detected at the chloroplast. In transient expression, several N-terminal deletions of the CP sequence, increasing in length, result in disruption of the domain functionality, markedly affecting intracellular localization. A yeast two-hybrid-based study using LoLV CP as bait identified several potentially interacting Arabidopsis host proteins, most of them with chloroplast-linked pathways. One of them, an ankyrin repeat protein, was studied in detail. The N. benthamiana homologue (NbANKr) targets chloroplasts, is able to co-localize with LoLV CP at chloroplast membranes in transient expression and shows a robust interaction with LoLV CP in vivo by BiFC, which has been confirmed by yeast two-hybrid data. Silencing NbANKr genes in N. benthamiana plants, prior to challenging with LoLV by mechanical inoculation, affects LoLV infection, significantly reducing the level of viral RNA in young leaves, compared to levels in control plants, and suggesting an inhibition of virus movement. Silencing of NbANKr has no obvious effect on plant phenotype, but is able to interfere with LoLV infection, opening the way for a new strategy for virus infection control.
ISSN:0022-1317
1465-2099
DOI:10.1099/jgv.0.001043