Short interfering RNA induced generation and translation of stable 5' mRNA cleavage intermediates

Singhania, Richa, Pavey, Sandra, Payne, Elizabeth, Gu, Wenyi, Clancy, Jennifer, Jubair, Luqman, Preiss, Thomas, Saunders, Nicholas and McMillan, Nigel A. J. (2016) Short interfering RNA induced generation and translation of stable 5' mRNA cleavage intermediates. Biochimica Et Biophysica Acta-Gene Regulatory Mechanisms, 1859 8: 1034-1042. doi:10.1016/j.bbagrm.2016.06.005

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Author Singhania, Richa
Pavey, Sandra
Payne, Elizabeth
Gu, Wenyi
Clancy, Jennifer
Jubair, Luqman
Preiss, Thomas
Saunders, Nicholas
McMillan, Nigel A. J.
Title Short interfering RNA induced generation and translation of stable 5' mRNA cleavage intermediates
Journal name Biochimica Et Biophysica Acta-Gene Regulatory Mechanisms   Check publisher's open access policy
ISSN 1874-9399
Publication date 2016-08-01
Year available 2016
Sub-type Article (original research)
DOI 10.1016/j.bbagrm.2016.06.005
Open Access Status File (Author Post-print)
Volume 1859
Issue 8
Start page 1034
End page 1042
Total pages 9
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Language eng
Subject 1304 Biophysics
1315 Structural Biology
1303 Biochemistry
1312 Molecular Biology
1311 Genetics
Abstract Sequence-specific degradation of homologous mRNA is the main mechanism by which short-interfering RNAs (siRNAs) suppress gene expression. Generally, it is assumed that the mRNA fragments resulting from Ago2 cleavage are rapidly degraded, thus making the transcript translation-incompetent. However, the molecular mechanisms involved in the post-cleavage mRNA decay are not completely understood and the fate of cleavage intermediates has been poorly studied. Using specific siRNAs and short-hairpin RNAs (shRNAs) we show that the 5′ and 3′ mRNA cleavage fragments of human papilloma virus type 16 (HPV-16) E6/7 mRNA, over-expressed in cervical malignancies, are unevenly degraded. Intriguingly, the 5′ mRNA fragment was more abundant and displayed a greater stability than the corresponding 3′ mRNA fragment in RNAi-treated cells. Further analysis revealed that the 5′ mRNA fragment was polysome-associated, indicating its active translation, and this was further confirmed by using tagged E7 protein to show that C-terminally truncated proteins were produced in treated cells. Overall, our findings provide new insight into the degradation of siRNA-targeted transcripts and show that RNAi can alter protein expression in cells as a result of preferential stabilization and translation of the 5′ cleavage fragment. These results challenge the current model of siRNA-mediated RNAi and provide a significant step forward towards understanding non-canonical pathways of siRNA gene silencing.
Keyword siRNA
mRNA cleavage
mRNA decay
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
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