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

Thiel, Volker; Siddell, Stuart G.

doi:10.1099/0022-1317-75-11-3041

Cited by 74 publications

(69 citation statements)

References 34 publications

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“…Except for the smallest mRNA, encoding just the N protein, they are all structurally polycistronic but express only their 5Ј-terminal open reading frame (ORF). The E protein, however, is translated from a downstream ORF by internal ribosome entry (3,44). Our quantitations of the synthesis of the S, N, M, and E proteins during infection show similar kinetics for all of them.…”

Section: Discussionmentioning

confidence: 65%

Characterization of the Coronavirus Mouse Hepatitis Virus Strain A59 Small Membrane Protein E

Raamsman

Locker

Hooge

et al. 2000

J Virol

175

204

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The small envelope (E) protein has recently been shown to play an essential role in the assembly of coronaviruses. Expression studies revealed that for formation of the viral envelope, actually only the E protein and the membrane (M) protein are required. Since little is known about this generally low-abundance virion component, we have characterized the E protein of mouse hepatitis virus strain A59 (MHV-A59), an 83-residue polypeptide. Using an antiserum to the hydrophilic carboxy terminus of this otherwise hydrophobic protein, we found that the E protein was synthesized in infected cells with similar kinetics as the other viral structural proteins. The protein appeared to be quite stable both during infection and when expressed individually using a vaccinia virus expression system. Consistent with the lack of a predicted cleavage site, the protein was found to become integrated in membranes without involvement of a cleaved signal peptide, nor were any other modifications of the polypeptide observed. Immunofluorescence analysis of cells expressing the E protein demonstrated that the hydrophilic tail is exposed on the cytoplasmic side. Accordingly, this domain of the protein could not be detected on the outside of virions but appeared to be inside, where it was protected from proteolytic degradation. The results lead to a topological model in which the polypeptide is buried within the membrane, spanning the lipid bilayer once, possibly twice, and exposing only its carboxy-terminal domain. Finally, electron microscopic studies demonstrated that expression of the E protein in cells induced the formation of characteristic membrane structures also observed in MHV-A59-infected cells, apparently consisting of masses of tubular, smooth, convoluted membranes. As judged by their colabeling with antibodies to E and to Rab-1, a marker for the intermediate compartment and endoplasmic reticulum, the E protein accumulates in and induces curvature into these pre-Golgi membranes where coronaviruses have been shown earlier to assemble by budding.Coronaviruses, a family of viruses belonging to the newly established order of the Nidovirales (for reviews, see references 8 and 37) have enveloped virions containing a nonsegmented, plus-stranded RNA genome. The RNA is packaged by the nucleocapsid (N) protein into a helical nucleocapsid. The surrounding envelope contains three, and sometimes four, membrane proteins. The spike (S) protein, a type I glycoprotein, occurs as trimers that constitute the characteristic surface projections. These function primarily in virus entry, being responsible for binding to the receptor on the target cell and for mediating fusion of viral and cellular membranes. The membrane (M) protein is a triple-spanning glycoprotein. It is the most abundant envelope protein component having essential functions in virus assembly. The hemagglutinin-esterase protein is present in only a subset of coronaviruses. The type I glycoprotein occurs in virions in disulfide-linked homodimeric form. Its biological role in the vi...

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Section: Discussionmentioning

confidence: 65%

Characterization of the Coronavirus Mouse Hepatitis Virus Strain A59 Small Membrane Protein E

Raamsman

Locker

Hooge

et al. 2000

J Virol

175

204

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“…RNA sequences having IRES properties has been found in number of viruses including most, if not all, the members of the picornavirus family [8], in hepatitis C [9], murine hepatitis virus [10], pestivirus [11], Moloney mouse leukaemia virus [12] and Friend mouse leukaemia virus [13]. These structures have a more or less similar mechanism of action [14,15].…”

Section: Introductionmentioning

confidence: 99%

The RU5 (‘R’) region from human leukaemia viruses (HTLV‐1) contains an internal ribosome entry site (IRES)‐like sequence

Attal¹,

Théron²,

Taboit³

et al. 1996

FEBS Letters

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RNA fragments containing the complete R region and the beginning of the U5 region ('R') from the human T cell leukaemia virus 1 (HTLV-1) stimulated the translation of the second eistrons in bicistronic mRNAs. The 5' untranslated region from SV40 early genes (SU) which was unable to stimulate translation of second cistrons amplified markedly the internal ribosome entry site (IRES) effect of the HTLV-1 'R' fragments. The 'R' regions from HTLV-1 have therefore properties similar to internal ribosome entry sites (IRES) originally found in picornavirus. The beginning of the U5 region from HTLV-1 contains a polypyrimidine sequence which is known to play an essential role in the IRES activity in picornavirus. The same experiments carried out using the 'R' region from bovine leukaemia virus (BLV) showed that this sequence has at most a weak IRES effect. One retroviruses HTLV-1 and perhaps others contain therefore an IRES activity. Interestingly, the combined SU 'R' sequence worked efficiently with different cistrons, different promoters and in all tested cell lines, whereas the poliovirus IRES was active in CHO cells but not in the mouse mammary cell line HCll. The SU 'R' sequence may therefore preferably be used to generate active bicistronic mRNAs.

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“…IRES function depends on primary, secondary and tertiary RNA structure. Since their discovery in picornaviruses, such IRES elements have also been identified within a number of other viral and cellular mRNAs (BERLIOZ et al 1995;BERNSTEIN et al 1997;GAN and RHOADS 1996;IrZUKA et al 1995;Lru and INGLIS 1992;MACEJAK and SARNOW 1991;MAGA et al 1995;NANBRU et al 1997;OH et al 1992;PRATS et al 1992;SIMONS et al 1996;STONELEY et al 1998;TEERINK et al 1995;THIEL and SIDDELL 1994;THOMAS et al 1991;VAGNER et al 1995a,b;VERVER et al 1991;YE et al 1997).…”

mentioning

confidence: 95%

Internal Ribosome Entry Site-Mediated Translation in Hepatitis C Virus Replication

Rijnbrand

Lemon

2000

Current Topics in Microbiology and Immunology

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

Cited by 74 publications

References 34 publications

Characterization of the Coronavirus Mouse Hepatitis Virus Strain A59 Small Membrane Protein E

Characterization of the Coronavirus Mouse Hepatitis Virus Strain A59 Small Membrane Protein E

The RU5 (‘R’) region from human leukaemia viruses (HTLV‐1) contains an internal ribosome entry site (IRES)‐like sequence

Internal Ribosome Entry Site-Mediated Translation in Hepatitis C Virus Replication

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