title: Severe acute respiratory syndrome coronavirus group-specific open reading frames encode nonessential functions for replication in cell cultures and mice.
abstract: SARS coronavirus (SARS-CoV) encodes several unique group-specific open reading frames (ORFs) relative to other known coronaviruses. To determine the significance of the SARS-CoV group-specific ORFs in virus replication in vitro and in mice, we systematically deleted five of the eight group-specific ORFs, ORF3a, OF3b, ORF6, ORF7a, and ORF7b, and characterized recombinant virus replication and gene expression in vitro. Deletion of the group-specific ORFs of SARS-CoV, either alone or in various combinations, did not dramatically influence replication efficiency in cell culture or in the levels of viral RNA synthesis. The greatest reduction in virus growth was noted following ORF3a deletion. SARS-CoV spike (S) glycoprotein does not encode a rough endoplasmic reticulum (rER)/Golgi retention signal, and it has been suggested that ORF3a interacts with and targets S glycoprotein retention in the rER/Golgi apparatus. Deletion of ORF3a did not alter subcellular localization of the S glycoprotein from distinct punctuate localization in the rER/Golgi apparatus. These data suggest that ORF3a plays little role in the targeting of S localization in the rER/Golgi apparatus. In addition, insertion of the 29-bp deletion fusing ORF8a/b into the single ORF8, noted in early-stage SARS-CoV human and civet cat isolates, had little if any impact on in vitro growth or RNA synthesis. All recombinant viruses replicated to wild-type levels in the murine model, suggesting that either the group-specific ORFs play little role in in vivo replication efficiency or that the mouse model is not of sufficient quality for discerning the role of the group-specific ORFs in disease origin and development.
authors.affiliation: Department of Epidemiology, School of Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7435, USA.
authors.forename: Boyd
authors.initials: B
authors.lastname: Yount
authors.forename: Rhonda S
authors.initials: RS
authors.lastname: Roberts
authors.forename: Amy C
authors.initials: AC
authors.lastname: Sims
authors.forename: Damon
authors.initials: D
authors.lastname: Deming
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Frieman
authors.forename: Jennifer
authors.initials: J
authors.lastname: Sparks
authors.forename: Mark R
authors.initials: MR
authors.lastname: Denison
authors.forename: Nancy
authors.initials: N
authors.lastname: Davis
authors.forename: Ralph S
authors.initials: RS
authors.lastname: Baric
mesh_terms: D000818:Animals; D018352:Coronavirus Infections / Q000821:virology*; D051379:Mice; D016366:Open Reading Frames / Q000235:genetics; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D014779:Virus Replication / Q000502:physiology*; D000086082:Viroporin Proteins
chemical_list: D014764:Viral Proteins; D015678:Viral Structural Proteins; C515313:ORF3A protein, SARS coronavirus; D000086082:Viroporin Proteins