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Importance of Unique Secondary Structures in Genomic RNA1 3′ CITE in Blackcurrant Reversion Nepovirus Translation
Author(s) -
Baquero Galvis Laura D.,
Shields Elizabeth,
FilbinWong Megan E.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.525.4
Subject(s) - rna , translation (biology) , biology , eukaryotic translation , internal ribosome entry site , genetics , non coding rna , nucleic acid structure , nucleic acid secondary structure , computational biology , messenger rna , gene
Translation initiation and regulation commonly involve end‐to‐end interactions between proteins (initiation factors) bound to a 7‐methyl guanosine triphosphate cap at the 5′ end, and at a poly(A) tail, at the 3′ end of the messenger RNA. End‐to‐end communication is also prevalent in many RNA viruses, such as the Blackcurrant Reversion Nepovirus (BRV), which has two bipartite genomic RNAs (RNA1 and RNA2), both of which contain the 3′ poly(A) tail but lack the 5′ cap. The mechanism implemented by BRV during translation initiation is not clearly defined. In other cap‐less, positive‐sense RNA plant viruses, structured RNA in the 5′ and 3′ UTRs bind via proposed RNA‐RNA kissing‐loop interactions indicating that the recruitment of protein synthesis machinery is directly related to their primary, secondary and higher‐order structure. One form of non‐canonical (cap‐independent) translation involves structured RNA called the cap‐independent translation enhancers (CITEs). These components are proposed to bind to initiation factors as a 5′cap‐replacement and play an important role in the efficient translation of uncapped RNA genomes. Considering the end‐to‐end communication and recruitment of translation factors is dependent upon RNA structure, our main goal focuses on determining the secondary and possible tertiary structures of the 3′ CITE in the genomic RNAs found in the BRV, specifically RNA1. With this in mind, we have probed our purified RNA with a series of covalent nucleobase, sugar and backbone modifiers in addition to testing the function of a series of structural mutants via reporter assays. We found that the RNA1 CITE structure plays an important role in the process of viral protein synthesis by either facilitating end‐to‐end interaction with the 5′ UTR and/or directly recruiting important translation protein machinery. These findings shed light on structure‐based mechanisms used for end‐to‐end communication in non‐canonical mRNAs. Support or Funding Information MSU Denver Colorado‐Wyoming Alliance for Minority Participation Undergraduate Research Grant, MSU Denver Dean's Grant, MSU Denver Provost Grant This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .