Formation of poliomyelitis subviral particles in the yeast <i>Saccharomyces cerevisiae</i>

Jore, J.p.m.; Veldhuisen, G.; Kottenhagen, M.J.; Pouwels, Peter H.; Foriers, André; Rombaut, Bartholomeus; Boeyé, Albert

doi:10.1002/yea.320100706

Cited by 6 publications

(4 citation statements)

References 34 publications

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“…Consistent with these morphogenetic features observed in higher eukaryotic cells, expression in yeast of these viral major components reveals intracellular assembly of VLPs, which are purified after deliberate disruption of yeast cells (5,6,41,42). However, this preparation procedure for VLPs is inappropriate for enveloped viruses because the particles form only concomitant with budding from the host lipid membrane at which time they adopt the lipid envelope.…”

Section: Discussionmentioning

confidence: 77%

HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomyces cerevisiae

Sakuragi

Goto

Sano

et al. 2002

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

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Expression of retroviral Gag protein in yeast has previously shown Gag targeting to the plasma membrane but little or no production of Gag virus-like particles (VLPs). Here we show that, after removal of the cell wall, the expression of HIV type 1 Gag protein in Saccharomyces cerevisiae spheroplasts allowed simultaneous budding of VLPs from the plasma membrane. Our data show that (i) the VLPs released from yeast spheroplasts were spherical and had morphological features, such as membrane apposed electrondense layers, characteristic of the immature form of HIV particles; (ii) the VLPs were completely enclosed in the plasma membrane derived from yeast, which is denser than that of higher eukaryotic cells; (iii) the VLP Gag shells remained intact after treatment of nonionic detergent; and (iv) the VLPs were released soon after removal of the cell wall and accumulated up to 300 g͞liter of culture. Our results also show that VLP production was abolished by amino acid substitution of the Gag N-terminal myristoylglycine and impaired when Gag C-terminal deletions were extended beyond the nucleocapsid domain. These results were consistent with those obtained previously in higher eukaryotic expression systems, suggesting that similar Gag domains were used for VLP assembly. We suggest that the system described here offers significant advantages for studying host factors required for VLP budding. The system also may be available for production of vector virus-free VLPs for practical applications such as vaccine development.

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Section: Discussionmentioning

confidence: 77%

HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomyces cerevisiae

Sakuragi

Goto

Sano

et al. 2002

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

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“…As an alternative, therefore, virus may be internal- Since the early 1980s, various yeasts have been used as an expression system for the production of VLPs. Hepatitis B (53), poliovirus (18), and a range of PVs VLPs, including HPV6 (13,38,45,55), cottontail rabbit PV (16), HPV16 (39,44,45), HPV18 (13), and HPV11 (3,19,30), have been produced in yeasts. Yeasts have also been used to study the assembly of PV VLPs and the efficiency of VLP production.…”

Section: Discussionmentioning

confidence: 99%

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

Zhao

Frazer

2002

J Virol

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We recently demonstrated that Saccharomyces cerevisiae protoplasts can take up bovine papillomavirus type 1 (BPV1) virions and that viral episomal DNA is replicated after uptake. Here we demonstrate that BPV virus-like particles are assembled in infected S. cerevisiae cultures from newly synthesized capsid proteins and also package newly synthesized DNA, including full-length and truncated viral DNA and S. cerevisiae-derived DNA. Virus particles prepared in S. cerevisiae are able to convey packaged DNA to Cos1 cells and to transform C127 cells. Infectivity was blocked by antisera to BPV1 L1 but not antisera to BPV1 E4. We conclude that S. cerevisiae is permissive for the replication of BPV1 virus.Papillomaviruses (PVs) are exclusively epitheliotropic viruses and have evolved a unique replication strategy that depends upon the differentiation program of keratinocytes (15). Though transient viral episome replication can occur in a number of in vitro cells (37), only keratinocytes, or cells with the potential for squamous maturation, can be productively infected, since viral capsid proteins are synthesized and virions are assembled only in terminally differentiated keratinocytes. PV capsid proteins, expressed in mammalian cells (61), insect cells (22), and E. coli (20,27), can be used to study virion assembly and DNA encapsidation (43,44,52,(57)(58)(59)(60). However, there remain large gaps in the understanding of PV life cycle. Kreider et al. (24) first reported the use of athymic mouse xenograft culture to produce infectious human PV type 11 (HPV11) in vivo. In vitro raft culture systems have allowed differentiation-specific viral amplification, late gene expression, and virion morphogenesis for HPV31 (9, 46) and other PV types (2, 34). Recently, infectious particles have been produced (2,8,31,35,40), although the viral yield is generally small compared to input virions. However, only a small number of HPV types can be successfully grown in athymic and scid mouse xenograft systems or raft culture systems (55), and propagation of large numbers of viral particles in vitro is yet to be achieved (2).Lambert et al. (26) first used the S. cerevisiae system to study the expression and function of the bovine PV type 1 (BPV1) E2 gene. Dostatni et al. (5) used S. cerevisiae to express fulllength BPV1 E2 protein and assayed in vitro its capacity to modulate transcription. Prakash et al. (41) reported that BPV1 E2 protein regulates viral transcription by binding as a dimer to the DNA sequence ACCGN4CGGT. According to previous studies of viral DNA replication in yeast (21, 42), the basic requirements for viral cis and trans elements for episome replication are similar between S. cerevisiae and mammalian cells.We have recently observed that S. cerevisiae protoplasts, which have extensive endocytotic activity (10), can take up BPV1 virions, and the BPV1 episome can replicate (56). In the present study, we have studied whether S. cerevisiae exposed to PV virions can support production of infectious virions. MATERIALS AND METHO...

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“…In contrast, nonenveloped viruses have evolved alternative pathways for release of infectious virions from infected cells, often requiring cooperativity among viral structural, nonstructural, and host proteins for successful release from infected cells (47). Although it has been established that coexpression of enterovirus structural polyprotein P1 and nonstructural polyprotein 3CD leads to the successful production of virions in vitro (20,(48)(49)(50) and protective immune responses in vivo (12,(21)(22)(23)(24)51), the necessity of this coexpression strategy had yet to be tested in the context of mRNA vaccines. We evaluated three alternative strategies, including secretion of VP1 and cytosolic expression of P1 polyprotein or P1 subunits, the latter mediated by ribosomal skipping motifs and recently described to produce VLPs in vitro (52).…”

Section: Discussionmentioning

confidence: 99%

“…To date, nucleic acid vaccine approaches for nonenveloped viruses, including enteroviruses, have focused on the expression of capsid subunits, including VP1, from DNA vaccine platforms (15)(16)(17). Recombinant protein approaches have established that coexpression of enterovirus P1 and 3CD polyproteins in yeast (11,(18)(19)(20) or insect cells (12,(21)(22)(23) can yield recombinant VLPs and that these antigens induce neutralizing antibody responses in animals. Subsequently, it was demonstrated that P1-3CD coexpression in vivo, achieved by an adenoviral vector, could similarly achieve neutralizing responses in mice (24).…”

Section: Introductionmentioning

confidence: 99%

A self-amplifying RNA vaccine prevents enterovirus D68 infection and disease in preclinical models

Warner,

Archer,

Park

et al. 2024

Sci. Transl. Med.

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The recent emergence and rapid response to severe acute respiratory syndrome coronavirus 2 was enabled by prototype pathogen and vaccine platform approaches, driven by the preemptive application of RNA vaccine technology to the related Middle East respiratory syndrome coronavirus. Recently, the National Institutes of Allergy and Infectious Diseases identified nine virus families of concern, eight enveloped virus families and one nonenveloped virus family, for which vaccine generation is a priority. Although RNA vaccines have been described for a variety of enveloped viruses, a roadmap for their use against nonenveloped viruses is lacking. Enterovirus D68 was recently designated a prototype pathogen within the family Picornaviridae of nonenveloped viruses because of its rapid evolution and respiratory route of transmission, coupled with a lack of diverse anti-enterovirus vaccine approaches in development. Here, we describe a proof-of-concept approach using a clinical stage RNA vaccine platform that induced robust enterovirus D68–neutralizing antibody responses in mice and nonhuman primates and prevented upper and lower respiratory tract infections and neurological disease in mice. In addition, we used our platform to rapidly characterize the antigenic diversity within the six genotypes of enterovirus D68, providing the necessary data to inform multivalent vaccine compositions that can elicit optimal breadth of neutralizing responses. These results demonstrate that RNA vaccines can be used as tools in our pandemic-preparedness toolbox for nonenveloped viruses.

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Formation of poliomyelitis subviral particles in the yeast Saccharomyces cerevisiae

Cited by 6 publications

References 34 publications

HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomyces cerevisiae

HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomyces cerevisiae

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

A self-amplifying RNA vaccine prevents enterovirus D68 infection and disease in preclinical models

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