Programmed ribosomal frameshifting in decoding the SARS-CoV genome.
Identifieur interne : 002572 ( PubMed/Checkpoint ); précédent : 002571; suivant : 002573Programmed ribosomal frameshifting in decoding the SARS-CoV genome.
Auteurs : Pavel V. Baranov [États-Unis] ; Clark M. Henderson ; Christine B. Anderson ; Raymond F. Gesteland ; John F. Atkins ; Michael T. HowardSource :
- Virology [ 0042-6822 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN viral (), ARN viral (génétique), Alignement de séquences, Conformation d'acide nucléique, Coronavirus (génétique), Données de séquences moléculaires, Décalage ribosomique du cadre de lecture (génétique), Gènes rapporteurs, Génome viral, Humains, Lignée cellulaire, Luciferases (génétique), Modèles moléculaires, Spectrométrie de masse ESI, Séquence nucléotidique, Virus du SRAS (génétique).
- MESH :
- génétique : ARN viral, Coronavirus, Décalage ribosomique du cadre de lecture, Luciferases, Virus du SRAS.
- ARN viral, Alignement de séquences, Conformation d'acide nucléique, Données de séquences moléculaires, Gènes rapporteurs, Génome viral, Humains, Lignée cellulaire, Modèles moléculaires, Spectrométrie de masse ESI, Séquence nucléotidique.
English descriptors
- KwdEn :
- Base Sequence, Cell Line, Coronavirus (genetics), Frameshifting, Ribosomal (genetics), Genes, Reporter, Genome, Viral, Humans, Luciferases (genetics), Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, RNA, Viral (chemistry), RNA, Viral (genetics), SARS Virus (genetics), Sequence Alignment, Spectrometry, Mass, Electrospray Ionization.
- MESH :
- chemical , chemistry : RNA, Viral.
- chemical , genetics : Luciferases, RNA, Viral.
- genetics : Coronavirus, Frameshifting, Ribosomal, SARS Virus.
- Base Sequence, Cell Line, Genes, Reporter, Genome, Viral, Humans, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Sequence Alignment, Spectrometry, Mass, Electrospray Ionization.
Abstract
Programmed ribosomal frameshifting is an essential mechanism used for the expression of orf1b in coronaviruses. Comparative analysis of the frameshift region reveals a universal shift site U_UUA_AAC, followed by a predicted downstream RNA structure in the form of either a pseudoknot or kissing stem loops. Frameshifting in SARS-CoV has been characterized in cultured mammalian cells using a dual luciferase reporter system and mass spectrometry. Mutagenic analysis of the SARS-CoV shift site and mass spectrometry of an affinity tagged frameshift product confirmed tandem tRNA slippage on the sequence U_UUA_AAC. Analysis of the downstream pseudoknot stimulator of frameshifting in SARS-CoV shows that a proposed RNA secondary structure in loop II and two unpaired nucleotides at the stem I-stem II junction in SARS-CoV are important for frameshift stimulation. These results demonstrate key sequences required for efficient frameshifting, and the utility of mass spectrometry to study ribosomal frameshifting.
DOI: 10.1016/j.virol.2004.11.038
PubMed: 15680415
Affiliations:
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pubmed:15680415Le document en format XML
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<front><div type="abstract" xml:lang="en">Programmed ribosomal frameshifting is an essential mechanism used for the expression of orf1b in coronaviruses. Comparative analysis of the frameshift region reveals a universal shift site U_UUA_AAC, followed by a predicted downstream RNA structure in the form of either a pseudoknot or kissing stem loops. Frameshifting in SARS-CoV has been characterized in cultured mammalian cells using a dual luciferase reporter system and mass spectrometry. Mutagenic analysis of the SARS-CoV shift site and mass spectrometry of an affinity tagged frameshift product confirmed tandem tRNA slippage on the sequence U_UUA_AAC. Analysis of the downstream pseudoknot stimulator of frameshifting in SARS-CoV shows that a proposed RNA secondary structure in loop II and two unpaired nucleotides at the stem I-stem II junction in SARS-CoV are important for frameshift stimulation. These results demonstrate key sequences required for efficient frameshifting, and the utility of mass spectrometry to study ribosomal frameshifting.</div>
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