Identification and characterization of a coronavirus packaging signal

Fosmire, Jennifer A.; Hwang, Kyongmin; Makino, Shinji

doi:10.1128/jvi.66.6.3522-3530.1992

Cited by 125 publications

(137 citation statements)

References 50 publications

(52 reference statements)

Supporting

Mentioning

132

Contrasting

Order By: Relevance

“…PSs of alpha-and gammacoronaviruses were first identified for TGEV and IBV (Escors et al, 2003;Penzes et al, 1994). MHV DI RNA studies revealed a 69-nt SL structure that was (i) located in the 3 0 region of ORF1b, (ii) confirmed to be required for DI RNA packaging, and (iii) shown to interact with the viral N protein (Fosmire et al, 1992;Molenkamp and Spaan, 1997;Woo et al, 1997). Subsequent studies indicated that a larger PS element and, possibly, additional factors are required for optimal packaging efficiency (Bos et al, 1997;Cologna and Hogue, 2000;Narayanan and Makino, 2001).…”

Section: Rna Elements Involved In Coronavirus Genome Packagingmentioning

confidence: 99%

Coronavirus cis-Acting RNA Elements

Madhugiri

Fricke

Marz

et al. 2016

Advances in Virus Research

101

111

View full text Add to dashboard Cite

Coronaviruses have exceptionally large RNA genomes of approximately 30 kilobases. Genome replication and transcription is mediated by a multisubunit protein complex comprised of more than a dozen virus-encoded proteins. The protein complex is thought to bind specific cis-acting RNA elements primarily located in the 5'- and 3'-terminal genome regions and upstream of the open reading frames located in the 3'-proximal one-third of the genome. Here, we review our current understanding of coronavirus cis-acting RNA elements, focusing on elements required for genome replication and packaging. Recent bioinformatic, biochemical, and genetic studies suggest a previously unknown level of conservation of cis-acting RNA structures among different coronavirus genera and, in some cases, even beyond genus boundaries. Also, there is increasing evidence to suggest that individual cis-acting elements may be part of higher-order RNA structures involving long-range and dynamic RNA-RNA interactions between RNA structural elements separated by thousands of nucleotides in the viral genome. We discuss the structural and functional features of these cis-acting RNA elements and their specific functions in coronavirus RNA synthesis.

show abstract

Section: Rna Elements Involved In Coronavirus Genome Packagingmentioning

confidence: 99%

Coronavirus cis-Acting RNA Elements

Madhugiri

Fricke

Marz

et al. 2016

Advances in Virus Research

101

111

View full text Add to dashboard Cite

show abstract

“…(Kim and Makino, 1995). After brief centrifugation of the supernatant, released viruses were partially purified using ultracentrifugation on a discontinuous sucrose gradient consisting of 60, 50, 30 and 20% sucrose, as described previously (Fosmire et al, 1992). After centrifugation, virus particles at the interface of 30 and 50% sucrose were collected, and further purified on a 10-60% continuous sucrose gradient at 26,000 rpm for 18 h at 4 • C in a Beckman SW28 rotor.…”

Section: Labeling Of Viral Proteins and Purification Of Virusesmentioning

confidence: 99%

“…In MHV dependency upon specific and selective packaging of intracellular genomic RNP complex is mediated by the selective interaction of one viral protein and one RNA element. The protein involved in packaging is the M protein (Narayanan et al, 2000), and the RNA element is a 190 nt-long RNA packaging signal (Narayanan and Makino, 2001b), which is present only in mRNA 1 but lacking in subgenomic mRNAs (Fosmire et al, 1992;van der Most et al, 1991). The interaction between M protein and the packaging signal leads to the subsequent packaging of only the genomic RNP complex from a pool of intracellular MHV RNP complexes (Narayanan and Makino, 2001b).…”

Section: Introductionmentioning

confidence: 99%

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

“…The most 3%-end region of MIGCAT came from the very 3%-end of the 0.46-kb long MHV-JHM genomic RNA. MIGCAT DI RNA contained MHV-JHM DI RNA cis-acting replication signals (Kim et al, 1993;Kim and Makino, 1995;Lin and Lai, 1993), a packaging signal (van der Most et al, 1991;Fosmire et al, 1992;Woo et al, 1997), and an intergenic sequence. We expected MIGCAT DI RNA to replicate and transcribe subgenomic DI RNA in MHV-infected cells.…”

mentioning

confidence: 99%

Importance of coronavirus negative-strand genomic RNA synthesis prior to subgenomic RNA transcription

Maeda

Makino

1998

Virus Research

View full text Add to dashboard Cite

The (-)-strand viral RNAs that result from after infection of cells with coronaviruses, which possess RNA genomes of message polarity, are genomic-sized and subgenomic-sized. Each of the (-)-strand subgenomic RNAs corresponds in size to each of the subgenomic mRNA species that are made in infected cells. We tested whether (-)-strand subgenomic RNAs might initially be synthesized from the input single-stranded (+)-strand genomic RNA prior to the production of subgenomic mRNAs. We used a mouse hepatitis virus (MHV) defective interfering (DI) RNA. from which subgenomic RNA was produced in DI RNA-replicating cells, because this DI RNA had a functional MHV intergenic region inserted in its interior. MHV samples containing the DI particles were irradiated with UV-light and then superinfected into cells that had been infected with MHV 4 h prior to superinfection. Northern blot analysis of intracellular RNAs that were extracted 3 h after superinfection showed that genomic DI RNA and subgenomic DI RNA had similar UV-target sizes, indicating that (-)-strand genomic DI RNA synthesis from input genomic DI RNA probably occurred prior to the subgenomic-size DI RNA synthesis. We discuss why, in the course of coronavirus transcription, (-)-strand genomic-length coronavirus RNA synthesis might occur before subgenomic-sized RNAs of either polarity are made.

show abstract

Identification and characterization of a coronavirus packaging signal

Cited by 125 publications

References 50 publications

Coronavirus cis-Acting RNA Elements

Coronavirus cis-Acting RNA Elements

Characterization of N protein self-association in coronavirus ribonucleoprotein complexes

Importance of coronavirus negative-strand genomic RNA synthesis prior to subgenomic RNA transcription

Contact Info

Product

Resources

About