Purification of Virus-like Particles of Recombinant Human Papillomavirus Type 11 Major Capsid Protein L1 from Saccharomyces cerevisiae

Cook, James C.; Joyce, Joseph G.; George, Hugh A.; Schultz, Loren D.; Hurni, William M.; Jansen, Kathrin U.; Hepler, Robert; Ip, Charlotte C. Yu; Lowe, Robert S.; Keller, Paul M.; Lehman, E. Dale

doi:10.1006/prep.1999.1155

Cited by 93 publications

(69 citation statements)

References 26 publications

(18 reference statements)

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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%

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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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Section: Discussionmentioning

confidence: 99%

“…After 4 days, fractions of VLP density were prepared as described above, dialyzed against PBS, and incubated with 10 U of DNase I in 10ϫ DNase buffer at 37°C for 30 min. Fractions were mixed with 50 l of 3 N NaOH and boiled for 5 min, and the amount of incorporated 3 H was determined by liquid scintillation counting (Beckman LS 7000).…”

mentioning

confidence: 99%

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

Zhao

Frazer

2002

J Virol

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“…[25][26][27] Purification of the recombinant VLPs, and the disassembly/ reassembly process was previously described. 28,29 The protein concentration was determined with the Pierce bicinchoninic acid (BCA) protein assay method, and then verified with UV absorbance at 280 nm.…”

Section: Hpv 6 11 16 and 18 L1 Vlpsmentioning

confidence: 99%

Characterization of virus-like particles in GARDASIL® by cryo transmission electron microscopy

Zhao

Potter

Carragher

et al. 2013

Human Vaccines & Immunotherapeutics

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“…HPV-11 L1 VLP were a generous gift from Merck Sharp & Dohme (West Point, Pa.). The HPV-11 L1 major coat protein was produced by using a Saccharomyces cerevisiae expression system as previously described (10,27). A panel of 95 synthetic 15-mer peptides spanning the entire sequence of the HPV-11 L1 VLP was synthesized by Eurogentec (Seraing, Belguim).…”

Section: Donorsmentioning

confidence: 99%

Analysis of CD4 ⁺ T-Cell Responses to Human Papillomavirus (HPV) Type 11 L1 in Healthy Adults Reveals a High Degree of Responsiveness and Cross-Reactivity with Other HPV Types

Williams

Hart

Wang

et al. 2002

J Virol

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Human papillomavirus type 11 (HPV-11؉ T-cell responses were cross-reactive. The degree of cross-reactivity was inversely related to the degree of L1 sequence diversity between these viruses. Finally, responses to HPV-11 L1 peptides were elicited from ex vivo CD45RO؉ peripheral blood mononuclear cells, demonstrating that recognition of HPV-11 was a specific memory response and not due to in vitro selection during tissue culture. This is the first study of CD4 ؉ T-cell responses to HPV-11 in healthy subjects and demonstrates marked cross-reactivity with other skin and genital HPV types. This cross-reactivity may be of significance for vaccine strategies against HPV-associated clinical diseases.

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Purification of Virus-like Particles of Recombinant Human Papillomavirus Type 11 Major Capsid Protein L1 from Saccharomyces cerevisiae

Cited by 93 publications

References 26 publications

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

Characterization of virus-like particles in GARDASIL® by cryo transmission electron microscopy

Analysis of CD4 ⁺ T-Cell Responses to Human Papillomavirus (HPV) Type 11 L1 in Healthy Adults Reveals a High Degree of Responsiveness and Cross-Reactivity with Other HPV Types

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Purification of Virus-like Particles of Recombinant Human Papillomavirus Type 11 Major Capsid Protein L1 from Saccharomyces cerevisiae

Cited by 93 publications

References 26 publications

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

Saccharomyces cerevisiaeIs Permissive for Replication of Bovine Papillomavirus Type 1

Characterization of virus-like particles in GARDASIL® by cryo transmission electron microscopy

Analysis of CD4 + T-Cell Responses to Human Papillomavirus (HPV) Type 11 L1 in Healthy Adults Reveals a High Degree of Responsiveness and Cross-Reactivity with Other HPV Types

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Analysis of CD4 ⁺ T-Cell Responses to Human Papillomavirus (HPV) Type 11 L1 in Healthy Adults Reveals a High Degree of Responsiveness and Cross-Reactivity with Other HPV Types