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Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus
The unique region of mRNA 5 of murine hepatitis virus contains two open reading frames, ORF 5a and ORF 5b. The downstream ORF 5b encodes the envelope (E) protein, an integral membrane protein of the virus. We have shown previously that the expression of ORF 5b is mediated by the internal entry of ri...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer-Verlag
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087331/ https://www.ncbi.nlm.nih.gov/pubmed/10416375 http://dx.doi.org/10.1007/s007050050556 |
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author | Jendrach, M. Thiel, V. Siddell, S. |
author_facet | Jendrach, M. Thiel, V. Siddell, S. |
author_sort | Jendrach, M. |
collection | PubMed |
description | The unique region of mRNA 5 of murine hepatitis virus contains two open reading frames, ORF 5a and ORF 5b. The downstream ORF 5b encodes the envelope (E) protein, an integral membrane protein of the virus. We have shown previously that the expression of ORF 5b is mediated by the internal entry of ribosomes. In the experiments reported here, we have used the in vitro translation of synthetic mRNAs to identify the region of mRNA 5 that mediates internal ribosome entry. Our results show that the 5′ border of the MHV mRNA 5 IRES element is located between nucleotides 227 and 244 in ORF 5a, while the 3′ border is located between nucleotides 140 and 172 in ORF 5b. The MHV mRNA 5 IRES element, therefore, contains not more than 280 nucleotides and encompasses the ORF 5b initiation codon. As evidenced by electrophoretic mobility shift assays, the IRES element of mRNA 5 interacts specifically with protein factors present in an L-cell lysate. |
format | Online Article Text |
id | pubmed-7087331 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Springer-Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-70873312020-03-23 Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus Jendrach, M. Thiel, V. Siddell, S. Arch Virol Article The unique region of mRNA 5 of murine hepatitis virus contains two open reading frames, ORF 5a and ORF 5b. The downstream ORF 5b encodes the envelope (E) protein, an integral membrane protein of the virus. We have shown previously that the expression of ORF 5b is mediated by the internal entry of ribosomes. In the experiments reported here, we have used the in vitro translation of synthetic mRNAs to identify the region of mRNA 5 that mediates internal ribosome entry. Our results show that the 5′ border of the MHV mRNA 5 IRES element is located between nucleotides 227 and 244 in ORF 5a, while the 3′ border is located between nucleotides 140 and 172 in ORF 5b. The MHV mRNA 5 IRES element, therefore, contains not more than 280 nucleotides and encompasses the ORF 5b initiation codon. As evidenced by electrophoretic mobility shift assays, the IRES element of mRNA 5 interacts specifically with protein factors present in an L-cell lysate. Springer-Verlag 2014-05-18 1999 /pmc/articles/PMC7087331/ /pubmed/10416375 http://dx.doi.org/10.1007/s007050050556 Text en © Springer-Verlag 1999 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Article Jendrach, M. Thiel, V. Siddell, S. Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus |
title | Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus |
title_full | Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus |
title_fullStr | Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus |
title_full_unstemmed | Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus |
title_short | Characterization of an internal ribosome entry site within mRNA 5 of murine hepatitis virus |
title_sort | characterization of an internal ribosome entry site within mrna 5 of murine hepatitis virus |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087331/ https://www.ncbi.nlm.nih.gov/pubmed/10416375 http://dx.doi.org/10.1007/s007050050556 |
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