Complementation of P37 (F13L gene) knock-out in vaccinia virus by a cell line expressing the gene constitutively.

Borrego, Belén; Lorenzo, Marı́a M.; Blasco, Rafael

doi:10.1099/0022-1317-80-2-425

Cited by 19 publications

(15 citation statements)

References 27 publications

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“…In agreement with this idea, a recent report showed that retention of a significant fraction of the B5R protein in the endoplasmic reticulum does not affect the wrapping process but rather leads to a reduction in the amount of B5R incorporated into EEV (26). Several of the EEV-specific proteins have been expressed previously in transfected cells (7,19,38,40). In general, our results using the SFV expression system are in good agreement with these studies.…”

Section: Vol 74 2000 Expression Of Vaccinia Virus Eev Envelope Protsupporting

confidence: 92%

Intracellular Localization of Vaccinia Virus Extracellular Enveloped Virus Envelope Proteins Individually Expressed Using a Semliki Forest Virus Replicon

Lorenzo¹,

Galindo²,

Griffiths³

et al. 2000

J Virol

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The extracellular enveloped virus (EEV) form of vaccinia virus is bound by an envelope which is acquired by wrapping of intracellular virus particles with cytoplasmic vesicles containing trans-Golgi network markers. Six virus-encoded proteins have been reported as components of the EEV envelope. Of these, four proteins (A33R, A34R, A56R, and B5R) are glycoproteins, one (A36R) is a nonglycosylated transmembrane protein, and one (F13L) is a palmitylated peripheral membrane protein. During infection, these proteins localize to the Golgi complex, where they are incorporated into infectious virus that is then transported and released into the extracellular medium. We have investigated the fates of these proteins after expressing them individually in the absence of vaccinia infection, using a Semliki Forest virus expression system. Significant amounts of proteins A33R and A56R efficiently reached the cell surface, suggesting that they do not contain retention signals for intracellular compartments. In contrast, proteins A34R and F13L were retained intracellularly but showed distributions different from that of the normal infection. Protein A36R was partially retained intracellularly, decorating both the Golgi complex and structures associated with actin fibers. A36R was also transported to the plasma membrane, where it accumulated at the tips of cell projections. Protein B5R was efficiently targeted to the Golgi region. A green fluorescent protein fusion with the last 42 C-terminal amino acids of B5R was sufficient to target the chimeric protein to the Golgi region. However, B5R-deficient vaccinia virus showed a normal localization pattern for other EEV envelope proteins. These results point to the transmembrane or cytosolic domain of B5R protein as one, but not the only, determinant of the retention of EEV proteins in the wrapping compartment.

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Section: Vol 74 2000 Expression Of Vaccinia Virus Eev Envelope Protsupporting

confidence: 92%

Intracellular Localization of Vaccinia Virus Extracellular Enveloped Virus Envelope Proteins Individually Expressed Using a Semliki Forest Virus Replicon

Lorenzo¹,

Galindo²,

Griffiths³

et al. 2000

J Virol

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“…Finally, the fusion protein F13-GFP showed perinuclear staining and diffuse cytoplasmic localization. The intracellular localization of these proteins was in general agreement to that previously reported (5,15,19,21,35), indicating that the tags used did not have a generalized effect on their distribution. From the above-described results, we concluded that these proteins, except GFP-tagged F12, showed at least some degree of accumulation in areas close to the nucleus (Fig.…”

supporting

confidence: 88%

“…In addition, several examples of modulation in the distribution of certain IEV envelope proteins have been reported. For instance, the distribution of F13 and its palmitylation are modified by infection (5). Also, the distribution of B5 is modified by coexpression of F13 (15), and the incorporation of A36 into IEV membranes has been shown to depend on the expression of A33 (45).…”

Section: Discussionmentioning

confidence: 99%

Interaction between Vaccinia Virus Extracellular Virus Envelope A33 and B5 Glycoproteins

Perdiguero¹,

Blasco²

2006

J Virol

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The extracellular form of vaccinia virus acquires its outer envelope by wrapping with cytoplasmic membranes that contain at least seven virus-encoded proteins, of which four are glycoproteins. We searched for interactions between the vaccinia virus A33 glycoprotein and proteins A34, A36, B5, F12, and F13. First, when myc epitope-tagged A33 was expressed in combination with other envelope proteins, A33 colocalized with B5 and A36, suggesting that direct A33-B5 and A33-A36 interactions occur in the absence of infection. A recombinant vaccinia virus (vA33Rmyc) was constructed by introduction of the myc-tagged A33 version (A33myc) into A33-deficient vaccinia virus. A33myc partially restored plaque formation and colocalized with enveloped virions in infected cells. Coimmunoprecipitation experiments with extracts of vA33Rmyc-infected cells confirmed the existence of a physical association of A33 with A36 and B5. Of these, the A33-B5 interaction is a novel finding, whereas the interaction between A33 and A36 has been previously characterized. Vaccinia virus is the most studied member of the family Poxviridae, which includes structurally large and genetically complex DNA viruses that replicate and assemble in the cytoplasm of infected cells. Vaccinia virus particles are assembled and released through different stages, including those of the noninfectious immature virus (IV), the intracellular mature virus (IMV), the intracellular enveloped virus (IEV), the cellassociated enveloped virus (CEV), and the extracellular enveloped virus (EEV) (24,36,37). IMV particles are cytosolic and remain intracellular until the cells are lysed. Some IMVs leave the assembly areas in a microtubule-dependent movement and become wrapped by a double layer of intracellular membranes derived from the early endosomes (38, 39) or trans-Golgi network (34) to form IEV. Subsequently, IEVs move to the cell surface (again requiring microtubules), where the outer membrane fuses with the plasma membrane, thus generating enveloped virions, with an extra membrane with respect to IMV, at the cell surface. Particles that remain attached to the plasma membrane are termed CEV, while those released are called EEV (2). In cell culture, enveloped virions (CEV plus EEV) are responsible for virus transmission, since mutants that form normal amounts of IMV but are blocked in virus wrapping are impaired in transmission (1). Specific roles in virus transmission have been assigned to CEV and EEV. CEV particles can induce the formation of actin tails, which are important for efficient cell-to-cell spread. Conversely, EEVs mediate longrange dissemination of virus (2, 6, 28).Seven virus-encoded IEV envelope proteins have been described and characterized. The A33 (29), A34 (7), B5 (9, 16), F13 (14), and A56 (35) proteins are present in IEV, CEV, and EEV, although approximately one-third of EEV particles lack A56 (20). Proteins A36 and F12 are present in IEV but absent from CEV and EEV envelopes (26,39,40,46). The proteins present in the IEV and EEV envelopes have been sho...

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“…It has been previously reported that F13L-deleted viruses are "severely attenuated" (25,26), although the details of those studies have not been presented. Nude and SCID mice were susceptible to intranasal VV-WR challenge even at doses as low as 100 pfu, but survived a F13L-KO challenge with 10 6 pfu and showed no evidence of disease during the 2-month monitoring period.…”

Section: Discussionmentioning

confidence: 99%

Efficacy of ST-246 versus lethal poxvirus challenge in immunodeficient mice

Grosenbach

Berhanu

King

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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The threat of smallpox as a bioweapon and the emerging threat of human monkeypox, among other poxviral diseases, highlight the need for effective poxvirus countermeasures. ST-246, which targets the F13L protein in vaccinia virus and its homologs in other orthopoxvirus species, provides full protection from lethal poxviral disease in numerous animal models and seems to be safe in humans. All previous evaluations of ST-246 efficacy have been in immunocompetent animals. However, the risk of severe poxviral disease is greater in immunodeficient hosts. Here we report on the efficacy of ST-246 in preventing or treating lethal poxviral disease in immunodeficient mice. After lethal challenge with the Western Reserve strain of vaccinia, Nude, SCID, and J H knockout mice additionally depleted of CD4 + and CD8 + T cells were not fully protected by ST-246, although survival was significantly extended. However, CD4 + T cell deficient, CD8 + T cell deficient, J H knockout, and J H knockout mice also deficient for CD4 + or CD8 + T cells survived lethal challenge when treated with ST-246 starting on the day of challenge. Delaying treatment until 72 h after infection reduced ST-246 efficacy in some models but provided full protection from lethal challenge in most. These findings suggest that ST-246 may be effective in controlling smallpox or other pathogenic orthopoxviruses in some immunodeficient human populations for whom the vaccine is contraindicated.antiviral | immunodeficiency | ST-246 | smallpox | orthopoxvirus

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Complementation of P37 (F13L gene) knock-out in vaccinia virus by a cell line expressing the gene constitutively.

Abstract: Vaccinia virus produces two different infectious forms, intracellular mature virus (IMV) and extracellular enveloped virus (EEV

Cited by 19 publications

References 27 publications

Intracellular Localization of Vaccinia Virus Extracellular Enveloped Virus Envelope Proteins Individually Expressed Using a Semliki Forest Virus Replicon

Intracellular Localization of Vaccinia Virus Extracellular Enveloped Virus Envelope Proteins Individually Expressed Using a Semliki Forest Virus Replicon

Interaction between Vaccinia Virus Extracellular Virus Envelope A33 and B5 Glycoproteins

Efficacy of ST-246 versus lethal poxvirus challenge in immunodeficient mice

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