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Title

A chloroplast protein binds a viroid RNA in vivo and facilitates its hammerhead-mediated self-cleavage

AuthorsDaròs Arnau, José Antonio CSIC ORCID ; Flores, Ricardo CSIC ORCID
KeywordsAvocado sunblotch viroid
Catalytic RNAs
Hammerhead ribozymes
RNA-binding proteins
RNA chaperones
Issue Date2002
PublisherNature Publishing Group
CitationEMBO Journal 21(4): 749–759 (2002)
AbstractViroids, small single-stranded circular RNAs (246–401 nucleotides), do not have mRNA capacity and must recruit host proteins to assist in the steps of their biological cycle. The nature of these cellular factors is poorly understood due to a lack of reliable experimental approaches. Here, to screen for host proteins interacting with viroid RNAs in vivo, we UV-irradiated avocado leaves infected with avocado sunblotch viroid (ASBVd), the type member of chloroplast viroids containing hammerhead ribozymes. This resulted in the detection of several ASBVd–host protein adducts. Tandem mass spectrometry analysis of the most abundant cross-linked species identified the protein component as two closely related chloroplast RNA-binding proteins (PARBP33 and PARBP35) of a family whose members previously have been shown to be involved in stabilization, maturation and editing of chloroplast transcripts. PARBP33 behaves as an RNA chaperone that stimulates in vitro the hammerhead-mediated self-cleavage of the multimeric ASBVd transcripts that result from rolling circle replication, indicating that this reaction, despite its RNA-based mechanism, is facilitated by proteins. The structural and functional parallelism between PARBP33 and PARBP35, and some proteins involved in viral RNA replication, indicates that viroids and RNA viruses recruit similar host proteins for their replication.
DescriptionFinal full-text version of the paper available at http://www.nature.com/emboj/index.html
URIhttp://hdl.handle.net/10261/3614
DOI10.1093/emboj/21.4.749
ISSN1460-2075
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