Abstract
Nonstructural Protein 1 (nsp1), generated by Severe Acute Respiratory Syndrome Coronavirus (SARS‐CoV), is responsible for host cell mRNA degradation via endonucleolytic cleavage. While the cellular mRNA is targeted for cleavage, few viral mRNAs are resistant. These mRNAs include mRNA from SARS‐CoV, hepatitis C virus and cricket paralysis virus. Other studies concluded that a stem‐loop forming sequence in the 5′‐UTR (untranslated region) may be responsible for this cleavage resistance. We designed multiple mutations in the UTR to partially or fully disrupt the stem‐loop, or reinstate a stem‐loop with mutated sequence. These RNAs were subjected to nsp1 mediated cleavage and RNA stability was quantified using reverse transcription followed by quantitative polymerase chain reaction. We used 18S ribosomal RNA as a control since it does not undergo nsp1 mediated cleavage. Surprisingly, our results show that a partially disrupted stem‐loop leads to enhanced stability of the RNA. In order to understand any sequence driven stability that arises due to stronger binding by nsp1, we are currently investigating RNA binding properties of nsp1.Support or Funding InformationSC INBRE faculty fellows awardSCICU summer fellows awardFurman fellows awardThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
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