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Volume 97, Issue 7

Parainfluenza virus chimeric mini-replicons indicate a novel regulatory element in the leader promoter Free

Abstract

Gene expression of paramyxoviruses is regulated by genome-encoded -acting elements; however, whether all the required elements for viral growth have been identified is not clear. Using a mini-replicon system, it has been shown that human parainfluenza virus type 2 (hPIV2) polymerase can recognize the promoter elements of parainfluenza virus type 5 (PIV5), but reporter activity is lower in this case. We constructed a series of luciferase-encoding chimeric PIV2/5 mini-genomes that are basically hPIV2, but whose leader (le), mRNA start signal and trailer sequence are partially replaced with those of PIV5. Studies of the chimeric PIV2/5 mini-replicons demonstrated that replacement of hPIV2 le with PIV5 le results in remarkably weak luciferase expression. Further mutagenesis identified the responsible region as positions 25–30 of the PIV5 le. Using recombinant hPIV2, the impact of this region on viral life cycles was assessed. Insertion of the mutation at this region facilitated viral growth, genomic replication and mRNA transcription at the early stage of infection, which elicited severe cell damage. In contrast, at the late infection stage it caused a reduction in viral transcription. Here, we identify a novel -acting element in the internal region of an le sequence that is involved in the regulation of polymerase, and which contributes to maintaining a balance between viral growth and cytotoxicity.

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Keyword(s): leader sequence , minireplicon and parainfluenza virus
© 2016 The Authors
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2016-07-01
2024-04-25
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Parainfluenza virus chimeric mini-replicons indicate a novel regulatory element in the leader promoter
J. Gen. Virol. 97, 1520 (2016); https://doi.org/10.1099/jgv.0.000479
/content/journal/jgv/10.1099/jgv.0.000479
/content/journal/jgv/10.1099/jgv.0.000479
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