Viral silencing suppressors and cellular proteins partner with plant RRP6-like exoribonucleases
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ORIGINAL PAPER
Viral silencing suppressors and cellular proteins partner with plant RRP6‑like exoribonucleases Miguel Ángel Freire1 Received: 20 February 2020 / Accepted: 2 June 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract RNA silencing and RNA decay are functionally interlaced, regulate gene expression and play a pivotal role in antiviral responses. As a counter-defensive strategy, many plant and mammalian viruses encode suppressors which interfere with both mechanisms. However, the protein interactions that connect these pathways remain elusive. Previous work reported that RNA silencing suppressors from different potyviruses, together with translation initiation factors EIF(iso)4E, interacted with the C-terminal region of the tobacco exoribonuclease RRP6-like 2, a component of the RNA decay exosome complex. Here, we investigate whether other viral silencing suppressors and cellular proteins might also bind RRP6-like exoribonucleases. A candidate search approach based on yeast two-hybrid protein interaction assays showed that three other unrelated viral suppressors, two from plant viruses and one from a mammalian virus, bound the C-terminus of the tobacco RRP6-like 2, the full-length of the Arabidopsis RRP6L1 protein and its C-terminal region. In addition, RRP6-like proteins were found to interact with members of the cellular double-stranded RNA-binding protein (DRB) family involved in RNA silencing. The C-terminal regions of RRP6L proteins are engaged in homotypic and heterotypic interactions and were predicted to be disordered. Collectively, these results suggest a protein interaction network that connects components of RNA decay and RNA silencing that is targeted by viral silencing suppressors. Keywords DRB proteins · Heat shock protein · RNA decay · RNase H-like · Viral silencing suppressor · Translation initiation factor EIF(iso)4E
Introduction RNA silencing and RNA decay are RNA-mediated gene inactivation pathways that regulate all levels of gene expression in eukaryotes and act as an antiviral immune system in plants. RNA silencing is a sequence-specific mechanism that relies on siRNAs and miRNAs assembling into ribonucleoprotein complexes. These complexes block transcription of Edited by Karel Petrzik. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11262-020-01775-z) contains supplementary material, which is available to authorized users. * Miguel Ángel Freire [email protected] 1
Instituto Multidisciplinario de Biología Vegetal (IMBIV), CONICET, Universidad Nacional de Córdoba (UNC), Av. Vélez Sarsfield 299, CC 495, 5000 Córdoba, Argentina
the homologous DNA loci by DNA methylation or histone modification (Transcriptional gene silencing, TGS); or cleave and/or repress the translation of the target mRNA (Post-transcriptional gene silencing, PTGS) [1]. RNA silencing is triggered by double-stranded RNA (dsRNA) precursors. Double-stranded RNA intermediates may be generated from different transcriptional unit
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