In Vitro synthesis of two polypeptides from a nonstructural gene of coronavirus mouse hepatitis virus strain A59

Budzilowicz, Carol; Weiss, Susan R.

doi:10.1016/0042-6822(87)90293-5

Cited by 55 publications

(52 citation statements)

References 32 publications

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“…The unique regions of the mRNAs encoding the N, M and S proteins of IBV, TGEV and MHV (see references in the section on virion proteins), the non-structural 15K protein of MHV and mRNAs 4 and 7 of TGEV each comprise only one ORF. In contrast, two ORFs are present in the unique region of the genomic RNA and of mRNA B of IBV, mRNAs 2 and 5 Budzilowicz & Weiss, 1987) of MHV and in mRNA 3 of TGEV , while three ORFs have been identified at the unique 5' end of IBV mRNA D (Boursnell et al, 1985b). These mRNAs may therefore be functionally polycistronic.…”

Section: Translation Strategymentioning

confidence: 99%

“…In vitro translation of RNA transcribed from cDNA containing both ORFs of MHV-A59 mRNA 5 indicated that the downstream reading frame is preferentially translated (Budzilowicz & Weiss, 1987). This ORF was shown to be expressed in infected cells (Leibowitz et al, 1988).…”

Section: Binns T D K Brown and S C Inglis Personal Communicationmentioning

confidence: 99%

“…A molar ratio of 1:2:2:7:11 has been estimated for D3 :S1 : indicate that the unique regions (enclosed by the junction sequences) of the corresponding mRNAs contain more than one ORF (for details see text). For IBV the data have been compiled from Boursnell et al (1987), for MHV from , , , Budzilowicz & Weiss (1987), Schmidtetal. (1987), Luytjesetal.…”

Section: Translation Strategymentioning

confidence: 99%

See 2 more Smart Citations

Coronaviruses: Structure and Genome Expression

Spaan

Cavanagh²,

Horzinek

1988

Journal of General Virology

503

485

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Section: Translation Strategymentioning

confidence: 99%

Section: Binns T D K Brown and S C Inglis Personal Communicationmentioning

confidence: 99%

Section: Translation Strategymentioning

confidence: 99%

See 1 more Smart Citation

Coronaviruses: Structure and Genome Expression

Spaan

Cavanagh²,

Horzinek

1988

Journal of General Virology

503

485

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“…These mRNAs appear to be functionally monocistronic. However, mRNA 1 and the subgenomic mRNA 5 are exceptions (Skinner et al, 1984;Budzilowicz & Weiss, 1987;Pachuk et al, 1989). The unique region of MHV mRNA 1 contains two large ORFs that constitute the coronavirus RNA polymerase locus.…”

Section: Introductionmentioning

confidence: 99%

Internal Ribosome Entry in the Coding Region of Murine Hepatitis Virus mRNA 5

Thiel

Siddell

1994

Journal of General Virology

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The unique region of murine hepatitis virus (MHV) mRNA 5 has two open reading frames, ORF 5a and ORF 5b, that encode small proteins of unknown function. In the experiments described here, we have used the in vitro translation of synthetic mRNAs to examine the expression of these ORFs. Our results show that a synthetic mRNA containing both ORFs is functionally bicistronic. More importantly, the expression of ORF 5b, but not ORF 5a, is maintained in a tricistronic mRNA containing an additional Y-proximal ORF. Thus, in the context of the MHV mRNA 5 unique region, the initiation of protein synthesis on ORF 5b can occur independently of ribosomes that enter from the 5' end of the mRNA. We conclude that the translation of ORF 5b is mediated by the internal entry of ribosomes.

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“…It could also form heterogeneous complexes with S, HE, and N proteins (36 -38). The interaction of these structural proteins with M may facilitate their assembly into the virion particles (2,36,39). However, M must also interact with E to be assembled into the virion, as expression of M alone could not induce the formation of VLP.…”

Section: Discussionmentioning

confidence: 99%

The Missing Link in Coronavirus Assembly

Lim

Liu

2001

Journal of Biological Chemistry

141

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One missing link in the coronavirus assembly is the physical interaction between two crucial structural proteins, the membrane (M) and envelope (E) proteins. In this study, we demonstrate that the coronavirus infectious bronchitis virus E can physically interact, via a putative peripheral domain, with M. Deletion of this domain resulted in a drastic reduction in the incorporation of M into virus-like particles. Immunofluorescent staining of cells coexpressing M and E supports that E interacts with M and relocates M to the same subcellular compartments that E resides in. E was retained in the pre-Golgi membranes, prior to being translocated to the Golgi apparatus and the secretory vesicles; M was observed to exhibit similar localization and translocation profiles as E when coexpressed with E. Deletion studies identified the C-terminal 6-residue RDKLYS as the endoplasmic reticulum retention signal of E, and site-directed mutagenesis of the ؊4 lysine residue to glutamine resulted in the accumulation of E in the Golgi apparatus. The third domain of E that plays a crucial role in virus budding is a putative transmembrane domain present at the N-terminal region, because deletion of the domain resulted in a free distribution of the mutant protein and in dysfunctional viral assembly.Morphogenesis and assembly of mammalian enveloped RNA viruses are complex processes. During these processes, the viral core, consisting of viral RNA and core proteins, becomes wrapped in a membranous structure (viral envelope) derived from host cell membranes to form virion particles. The assembly process, which is referred to as budding, usually occurs at either the plasma or intracellular membranes. The formation of viral core, envelope, and the assembly of virus particles would involve interaction between viral proteins and host membrane components, among viral structural proteins and between viral proteins and viral RNA. Characterization of these events at the molecular level has been greatly facilitated by the understanding that viral structural proteins may contain all of the necessary information to dictate the assembly process. In fact, it was observed that coexpression of viral structural proteins would result in the formation of virus-like particles (VLPs) 1 in many different viral systems (1-9). In this study, we exploit the coronavirus VLP system to study the interaction between two structural proteins during coronavirus assembly and the implication of this interaction in the assembly and release of coronavirus particles. Coronavirus is the largest RNA virus known so far. It has a positive-sense, single-strand RNA genome of 27-30 kilobases in length. Despite of the huge genome size, coronavirus typically contains four structural proteins, i.e. a type I spike (S) glycoprotein required for infectivity, a phosphorylated nucleocapsid (N) protein that interacts with the viral genome to form a helical core, a major type III integral membrane (M) protein, and a minor type III envelope (E) protein (10 -15). A fifth protein, the hemagglut...

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In Vitro synthesis of two polypeptides from a nonstructural gene of coronavirus mouse hepatitis virus strain A59

Cited by 55 publications

References 32 publications

Coronaviruses: Structure and Genome Expression

Coronaviruses: Structure and Genome Expression

Internal Ribosome Entry in the Coding Region of Murine Hepatitis Virus mRNA 5

The Missing Link in Coronavirus Assembly

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