Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
J Virol
2006 Feb 01;804:1653-61. doi: 10.1128/JVI.80.4.1653-1661.2006.
Show Gene links
Show Anatomy links
Site-directed mutagenesis of the Nidovirus replicative endoribonuclease NendoU exerts pleiotropic effects on the arterivirus life cycle.
Posthuma CC
,
Nedialkova DD
,
Zevenhoven-Dobbe JC
,
Blokhuis JH
,
Gorbalenya AE
,
Snijder EJ
.
???displayArticle.abstract???
The highly conserved NendoU replicative domain of nidoviruses (arteriviruses, coronaviruses, and roniviruses) belongs to a small protein family whose cellular branch is prototyped by XendoU, a Xenopus laevis endoribonuclease involved in nucleolar RNA processing. Recently, sequence-specific in vitro endoribonuclease activity was demonstrated for the NendoU-containing nonstructural protein (nsp) 15 of several coronaviruses. To investigate the biological role of this novel enzymatic activity, we have characterized a comprehensive set of arterivirus NendoU mutants. Deleting parts of the NendoU domain from nsp11 of equine arteritis virus was lethal. Site-directed mutagenesis of conserved residues exerted pleiotropic effects. In a first-cycle analysis, replacement of two conserved Asp residues in the C-terminal part of NendoU rendered viral RNA synthesis and virus production undetectable. In contrast, mutagenesis of other conserved residues, including two putative catalytic His residues that are absolutely conserved in NendoU and cellular homologs, produced viable mutants displaying reduced plaque sizes (20 to 80% reduction) and reduced yields of infectious progeny of up to 5 log units. A more detailed analysis of these mutants revealed a moderate reduction in RNA synthesis, with subgenomic RNA synthesis consistently being more strongly affected than genome replication. Our data suggest that the arterivirus nsp11 is a multifunctional protein with a key role in viral RNA synthesis and additional functions in the viral life cycle that are as yet poorly defined.
Bhardwaj,
The severe acute respiratory syndrome coronavirus Nsp15 protein is an endoribonuclease that prefers manganese as a cofactor.
2004, Pubmed
Bhardwaj,
The severe acute respiratory syndrome coronavirus Nsp15 protein is an endoribonuclease that prefers manganese as a cofactor.
2004,
Pubmed
Cavanagh,
Nidovirales: a new order comprising Coronaviridae and Arteriviridae.
1997,
Pubmed
Cowley,
Gill-associated virus of Penaeus monodon prawns: an invertebrate virus with ORF1a and ORF1b genes related to arteri- and coronaviruses.
2000,
Pubmed
Cowley,
Gill-associated nidovirus of Penaeus monodon prawns transcribes 3'-coterminal subgenomic mRNAs that do not possess 5'-leader sequences.
2002,
Pubmed
Feder,
Molecular phylogenetics of the RrmJ/fibrillarin superfamily of ribose 2'-O-methyltransferases.
2003,
Pubmed
Gioia,
Functional characterization of XendoU, the endoribonuclease involved in small nucleolar RNA biosynthesis.
2005,
Pubmed
,
Xenbase
Gorbalenya,
Big nidovirus genome. When count and order of domains matter.
2001,
Pubmed
Gorbalenya,
Coronavirus genome: prediction of putative functional domains in the non-structural polyprotein by comparative amino acid sequence analysis.
1989,
Pubmed
Ivanov,
Major genetic marker of nidoviruses encodes a replicative endoribonuclease.
2004,
Pubmed
Landt,
A general method for rapid site-directed mutagenesis using the polymerase chain reaction.
1990,
Pubmed
Laneve,
Purification, cloning, and characterization of XendoU, a novel endoribonuclease involved in processing of intron-encoded small nucleolar RNAs in Xenopus laevis.
2003,
Pubmed
,
Xenbase
Molenkamp,
The arterivirus replicase is the only viral protein required for genome replication and subgenomic mRNA transcription.
2000,
Pubmed
Pasternak,
Sequence requirements for RNA strand transfer during nidovirus discontinuous subgenomic RNA synthesis.
2001,
Pubmed
Sawicki,
Coronavirus transcription: a perspective.
2005,
Pubmed
Sawicki,
Coronaviruses use discontinuous extension for synthesis of subgenome-length negative strands.
1995,
Pubmed
Seybert,
A complex zinc finger controls the enzymatic activities of nidovirus helicases.
2005,
Pubmed
Seybert,
Biochemical characterization of the equine arteritis virus helicase suggests a close functional relationship between arterivirus and coronavirus helicases.
2000,
Pubmed
Snijder,
Unique and conserved features of genome and proteome of SARS-coronavirus, an early split-off from the coronavirus group 2 lineage.
2003,
Pubmed
Tijms,
A zinc finger-containing papain-like protease couples subgenomic mRNA synthesis to genome translation in a positive-stranded RNA virus.
2001,
Pubmed
Van Den Born,
Secondary structure and function of the 5'-proximal region of the equine arteritis virus RNA genome.
2004,
Pubmed
Wieringa,
Structural protein requirements in equine arteritis virus assembly.
2004,
Pubmed
Yan,
Assessment of putative protein targets derived from the SARS genome.
2003,
Pubmed
Zevenhoven-Dobbe,
Rescue of disabled infectious single-cycle (DISC) equine arteritis virus by using complementing cell lines that express minor structural glycoproteins.
2004,
Pubmed
Ziebuhr,
Virus-encoded proteinases and proteolytic processing in the Nidovirales.
2000,
Pubmed
Ziebuhr,
Molecular biology of severe acute respiratory syndrome coronavirus.
2004,
Pubmed
de Vries,
All subgenomic mRNAs of equine arteritis virus contain a common leader sequence.
1990,
Pubmed
de Vries,
Structural proteins of equine arteritis virus.
1992,
Pubmed
den Boon,
Equine arteritis virus is not a togavirus but belongs to the coronaviruslike superfamily.
1991,
Pubmed
van Dinten,
Proteolytic processing of the open reading frame 1b-encoded part of arterivirus replicase is mediated by nsp4 serine protease and Is essential for virus replication.
1999,
Pubmed
van Dinten,
The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis.
2000,
Pubmed
van Dinten,
An infectious arterivirus cDNA clone: identification of a replicase point mutation that abolishes discontinuous mRNA transcription.
1997,
Pubmed
van Marle,
Characterization of an equine arteritis virus replicase mutant defective in subgenomic mRNA synthesis.
1999,
Pubmed
van Marle,
Arterivirus discontinuous mRNA transcription is guided by base pairing between sense and antisense transcription-regulating sequences.
1999,
Pubmed
van Vliet,
Discontinuous and non-discontinuous subgenomic RNA transcription in a nidovirus.
2002,
Pubmed
van den Born,
Discontinuous subgenomic RNA synthesis in arteriviruses is guided by an RNA hairpin structure located in the genomic leader region.
2005,
Pubmed
van der Meer,
ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex.
1998,
Pubmed
von Grotthuss,
mRNA cap-1 methyltransferase in the SARS genome.
2003,
Pubmed