Repair and mutagenesis of the genome of a deletion mutant of the coronavirus mouse hepatitis virus by targeted RNA recombination

Koetzner, Cheri A.; Parker, Monica M.; Ricard, Cynthia S.; Sturman, Lawrence S.; Masters, Paul S.

doi:10.1128/jvi.66.4.1841-1848.1992

Cited by 131 publications

(213 citation statements)

References 34 publications

Supporting

Mentioning

207

Contrasting

Order By: Relevance

“…The plaque-cloned A59 strain of MHV (MHV-A59) (Lai et al, 1981) and its temperature-sensitive mutant Alb4 (Koetzner et al, 1992) were used for this study. MHV-A59 and Alb4 stock viruses were propagated in mouse DBT cells (Hirano et al, 1974) at 37 • C and 33 • C, respectively.…”

Section: Viruses and Cellsmentioning

confidence: 99%

“…We assumed that N proteins in MHV virion nucleocapsid were tightly associated with each other, and asked whether this putative tight N protein association actually was maintained after RNase A-treatment of the virion nucleocapsid had extensively degraded the RNA. MHV-A59 and its temperature-sensitive mutant Alb4 (Koetzner et al, 1992) were used to address this question. MHV N protein consists of three domains, each of which is connected by a short spacer sequence (Koetzner et al, 1992).…”

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

“…MHV-A59 and its temperature-sensitive mutant Alb4 (Koetzner et al, 1992) were used to address this question. MHV N protein consists of three domains, each of which is connected by a short spacer sequence (Koetzner et al, 1992). Alb4 replicates well at 33 • C, and its N protein has a deletion of 29-amino-acids within one of the spacer regions (Koetzner et al, 1992).…”

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

“…MHV N protein consists of three domains, each of which is connected by a short spacer sequence (Koetzner et al, 1992). Alb4 replicates well at 33 • C, and its N protein has a deletion of 29-amino-acids within one of the spacer regions (Koetzner et al, 1992). We raised an anti-peptide antibody, anti-N-spacer antibody, against part of the specific 29-amino-acid sequence, with the expectation that this antibody would recognize only the MHV-A59 N protein and not the Alb4 N protein.…”

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

“…We wanted to test whether the putative tight association of N proteins in the viral nucleocapsid was real enough to be maintained even after extensive degradation of the genomic RNA; hypothetically, RNase A-treatment of the virion nucleocapsid from coinfected cells followed by immunoprecipitation using anti-N-spacer antibody would likely result in the coimmunoprecipitation of Alb4 N protein with wt MHV-A59 N protein only if the two protein species had previously been associated with each other. Due to its deletion the Alb4 N protein migrates faster than MHV-A59 N protein in SDS-PAGE (Koetzner et al, 1992), which makes the identification of each N protein feasible. If the N protein monomers in the viral nucleocapsid are not associated with each other, then anti-N-spacer antibody most likely should immunoprecipitate only the MHV-A59 N protein from the same sample preparation.…”

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

See 4 more Smart Citations

Characterization of N protein self-association in coronavirus ribonucleoprotein complexes

2003

View full text Add to dashboard Cite

Mouse hepatitis virus (MHV) nucleocapsid (N) protein binds to the large, single-stranded, positive-sense viral genomic RNA to form a helical nucleocapsid structure in mature virions. In addition N protein binds the intracellular form of the genomic RNA, all of the MHV subgenomic mRNAs, and expressed non-MHV RNA transcripts to form ribonucleoprotein (RNP) complexes in infected cells. Among the intracellular viral RNP complexes, only the genomic RNP complex is packaged into virus particles. The present study demonstrated that N protein in the MHV virion nucleocapsid and in the intracellular genome-length RNP complex that bound to viral envelope M protein was tightly self-associated such that its association was retained even after extensive RNase A-treatment of the RNP complexes. The RNase A-resistant tight N protein association in the virion nucleocapsid was not mediated by an intermolecular disulfide bridge between N proteins. In contrast, N protein association in the majority of the intracellular RNP complexes was susceptible to RNase A-treatment. Because the RNP complexes that specifically interact with the M protein are selectively packaged into MHV particles, the present data suggested that there was a distinct difference between N protein association in viral genomic RNP complexes that undergo packaging into virus particles and the subgenomic RNP complexes that are not packaged into MHV particles.

show abstract

Section: Viruses and Cellsmentioning

confidence: 99%

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

Section: Experimental Approach For Analyzing N Protein Self-associatimentioning

confidence: 99%

See 3 more Smart Citations

Characterization of N protein self-association in coronavirus ribonucleoprotein complexes

2003

View full text Add to dashboard Cite

show abstract

Coronavirus Reverse Genetics

Armesto

Bentley

Bickerton

et al. 2012

Reverse Genetics of RNA Viruses

View full text Add to dashboard Cite

The Coronaviridae form part of the order Nidovirales, which comprises two subfamilies, the Coronavirinae and Torovirinae. There are three genera of coronaviruses, alpha-, beta-and gammacoronaviruses (Carstens, 2010), which were so named for their visual resemblance to the corona of the sun in negatively stained preparations (Figure 2.1) (Tyrrell et al., 1968). Representative members of each of the coronavirus genera are shown in Table 2.1. Possibly the most publicised coronavirus of recent years has been the human coronavirus SARS-CoV, which emerged in China in 2002 causing the severe acute respiratory syndrome epidemic (

show abstract

Recombination in large RNA viruses: Coronaviruses

Lai

1996

Seminars in Virology

View full text Add to dashboard Cite

Coronaviruses contain a very large RNA genome, which undergoes recombination at a very high frequency of nearly 25% for the entire genome. Recombination has been demonstrated to occur between viral genomes and between defective-interfering (DI) RNAs and viral RNA. It provides an evolutionary tool for both viral RNAs and DI RNA and may account for the diversity in the genomic structure of coronaviruses. The capacity of coronaviruses to undergo recombination may be related to its mRNA transcription mechanism, which involves discontinuous RNA synthesis, suggesting the nonprocessive nature of the viral polymerase. Recombination is used as a tool for the mutagenesis of viral genomic RNA.

show abstract

Repair and mutagenesis of the genome of a deletion mutant of the coronavirus mouse hepatitis virus by targeted RNA recombination

Cited by 131 publications

References 34 publications

Characterization of N protein self-association in coronavirus ribonucleoprotein complexes

Characterization of N protein self-association in coronavirus ribonucleoprotein complexes

Coronavirus Reverse Genetics

Recombination in large RNA viruses: Coronaviruses

Contact Info

Product

Resources

About