Genome structure and transcriptional regulation of human coronavirus NL63
<p>Abstract</p> <p>Background</p> <p>Two human coronaviruses are known since the 1960s: HCoV-229E and HCoV-OC43. SARS-CoV was discovered in the early spring of 2003, followed by the identification of HCoV-NL63, the fourth member of the coronaviridae family that infects...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2004-11-01
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| Series: | Virology Journal |
| Online Access: | http://www.virologyj.com/content/1/1/7 |
| _version_ | 1818209091397353472 |
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| author | Berkhout Ben Jebbink Maarten F Pyrc Krzysztof van der Hoek Lia |
| author_facet | Berkhout Ben Jebbink Maarten F Pyrc Krzysztof van der Hoek Lia |
| author_sort | Berkhout Ben |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Two human coronaviruses are known since the 1960s: HCoV-229E and HCoV-OC43. SARS-CoV was discovered in the early spring of 2003, followed by the identification of HCoV-NL63, the fourth member of the coronaviridae family that infects humans. In this study, we describe the genome structure and the transcription strategy of HCoV-NL63 by experimental analysis of the viral subgenomic mRNAs.</p> <p>Results</p> <p>The genome of HCoV-NL63 has the following gene order: 1a-1b-S-ORF3-E-M-N. The GC content of the HCoV-NL63 genome is extremely low (34%) compared to other coronaviruses, and we therefore performed additional analysis of the nucleotide composition. Overall, the RNA genome is very low in C and high in U, and this is also reflected in the codon usage. Inspection of the nucleotide composition along the genome indicates that the C-count increases significantly in the last one-third of the genome at the expense of U and G. We document the production of subgenomic (sg) mRNAs coding for the S, ORF3, E, M and N proteins. We did not detect any additional sg mRNA. Furthermore, we sequenced the 5' end of all sg mRNAs, confirming the presence of an identical leader sequence in each sg mRNA. Northern blot analysis indicated that the expression level among the sg mRNAs differs significantly, with the sg mRNA encoding nucleocapsid (N) being the most abundant.</p> <p>Conclusions</p> <p>The presented data give insight into the viral evolution and mutational patterns in coronaviral genome. Furthermore our data show that HCoV-NL63 employs the discontinuous replication strategy with generation of subgenomic mRNAs during the (-) strand synthesis. Because HCoV-NL63 has a low pathogenicity and is able to grow easily in cell culture, this virus can be a powerful tool to study SARS coronavirus pathogenesis.</p> |
| first_indexed | 2024-12-12T04:55:12Z |
| format | Article |
| id | doaj.art-3df89d24044143aba892a7d8595a787c |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-12T04:55:12Z |
| publishDate | 2004-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-3df89d24044143aba892a7d8595a787c2022-12-22T00:37:22ZengBMCVirology Journal1743-422X2004-11-0111710.1186/1743-422X-1-7Genome structure and transcriptional regulation of human coronavirus NL63Berkhout BenJebbink Maarten FPyrc Krzysztofvan der Hoek Lia<p>Abstract</p> <p>Background</p> <p>Two human coronaviruses are known since the 1960s: HCoV-229E and HCoV-OC43. SARS-CoV was discovered in the early spring of 2003, followed by the identification of HCoV-NL63, the fourth member of the coronaviridae family that infects humans. In this study, we describe the genome structure and the transcription strategy of HCoV-NL63 by experimental analysis of the viral subgenomic mRNAs.</p> <p>Results</p> <p>The genome of HCoV-NL63 has the following gene order: 1a-1b-S-ORF3-E-M-N. The GC content of the HCoV-NL63 genome is extremely low (34%) compared to other coronaviruses, and we therefore performed additional analysis of the nucleotide composition. Overall, the RNA genome is very low in C and high in U, and this is also reflected in the codon usage. Inspection of the nucleotide composition along the genome indicates that the C-count increases significantly in the last one-third of the genome at the expense of U and G. We document the production of subgenomic (sg) mRNAs coding for the S, ORF3, E, M and N proteins. We did not detect any additional sg mRNA. Furthermore, we sequenced the 5' end of all sg mRNAs, confirming the presence of an identical leader sequence in each sg mRNA. Northern blot analysis indicated that the expression level among the sg mRNAs differs significantly, with the sg mRNA encoding nucleocapsid (N) being the most abundant.</p> <p>Conclusions</p> <p>The presented data give insight into the viral evolution and mutational patterns in coronaviral genome. Furthermore our data show that HCoV-NL63 employs the discontinuous replication strategy with generation of subgenomic mRNAs during the (-) strand synthesis. Because HCoV-NL63 has a low pathogenicity and is able to grow easily in cell culture, this virus can be a powerful tool to study SARS coronavirus pathogenesis.</p>http://www.virologyj.com/content/1/1/7 |
| spellingShingle | Berkhout Ben Jebbink Maarten F Pyrc Krzysztof van der Hoek Lia Genome structure and transcriptional regulation of human coronavirus NL63 Virology Journal |
| title | Genome structure and transcriptional regulation of human coronavirus NL63 |
| title_full | Genome structure and transcriptional regulation of human coronavirus NL63 |
| title_fullStr | Genome structure and transcriptional regulation of human coronavirus NL63 |
| title_full_unstemmed | Genome structure and transcriptional regulation of human coronavirus NL63 |
| title_short | Genome structure and transcriptional regulation of human coronavirus NL63 |
| title_sort | genome structure and transcriptional regulation of human coronavirus nl63 |
| url | http://www.virologyj.com/content/1/1/7 |
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