Characterization of the gene encoding the A-type inclusion protein of camelpox virus and sequence comparison with other orthopoxviruses

Meyer, Hermann; Rziha, Hanns-Joachim

doi:10.1099/0022-1317-74-8-1679

Cited by 39 publications

(33 citation statements)

References 22 publications

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“…Apart from soluble exoantigens, peptides corresponding to parasite-derived iRBC plasma membrane-associated proteins were also identified in the extracellular supernatant. For example, a vacuolar protein sorting component, VPS45; a membrane-bound putative monocarboxylate transporter; LCCL domain-containing protein CCP1; a FIKK kinase (21); and a hypothetical protein with similarity to viral A type inclusion proteins (22) were identified along with a few hypothetical proteins (Table I). Predicted functions of the observed proteins are indicative of their secretory and/or surface-associated nature.…”

Section: De/ms and Lc-ms/ms Of P Falciparum Extracellular Se-mentioning

confidence: 99%

Proteome Analysis of Plasmodium falciparum Extracellular Secretory Antigens at Asexual Blood Stages Reveals a Cohort of Proteins with Possible Roles in Immune Modulation and Signaling

Singh

Mukherjee

Narayanasamy³

et al. 2009

Molecular & Cellular Proteomics

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The highly co-evolved relationship of parasites and their hosts appears to include modulation of host immune signals, although the molecular mechanisms involved in the host-parasite interplay remain poorly understood. Characterization of these key genes and their cognate proteins related to the host-parasite interplay should lead to a better understanding of this intriguing biological phenomenon. The malaria agent Plasmodium falciparum is predicted to export a cohort of several hundred proteins to remodel the host erythrocyte. However, proteins actively exported by the asexual intracellular parasite beyond the host red blood cell membrane (before merozoite egress) have been poorly investigated so far. Here we used two complementary methodologies, two-dimensional gel electrophoresis/MS and LC-MS/MS, to examine the extracellular secreted antigens at asexual blood stages of P. falciparum. We identified 27 novel antigens exported by P. falciparum in the culture medium of which some showed clustering with highly polymorphic genes on chromosomes, suggesting that they may encode putative antigenic determinants of the parasite. Immunolocalization of four novel secreted proteins confirmed their export beyond the infected red blood cell membrane. Of these, preliminary functional characterization of two novel (Sel1 repeat-containing) parasite proteins, PfSEL1 and PfSEL2 revealed that they down-regulate expression of cell surface Notch signaling molecules in host cells. Also a novel protein kinase (PfEK) and a novel protein phosphatase (PfEP) were found to, respectively, phosphorylate/dephosphorylate parasite-specific proteins in the extracellular culture supernatant. Our study thus sheds new light on malaria parasite extracellular secreted antigens of which some may be essential for parasite development and could constitute promising new drug targets. Molecular & Cellular Proteomics 8:2102-2118, 2009.

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Section: De/ms and Lc-ms/ms Of P Falciparum Extracellular Se-mentioning

confidence: 99%

Proteome Analysis of Plasmodium falciparum Extracellular Secretory Antigens at Asexual Blood Stages Reveals a Cohort of Proteins with Possible Roles in Immune Modulation and Signaling

Singh

Mukherjee

Narayanasamy³

et al. 2009

Molecular & Cellular Proteomics

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“…All CMLV isolates were initially confirmed by previously described 7,15 OPXV-specific, ATI gene-based PCR technique. Furthermore, for sequencing of the partial fragment of the C18L gene, extracted gDNA from all the CMLV isolates was subjected to amplification using 10 pmol of each primer (OPXV-specific C18L F and C18L R), 5 ml of 103 PCR buffer, 10 mmol/l each deoxyribonucleotide triphosphate (dNTP), and 0.5 IU of Taq DNA polymerase c in a thermal cycler.…”

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confidence: 99%

“…For the detection of OPXV infections in camels, Southern blot hybridization, REA, mapping of viral DNA, and A type inclusion (ATI ) genespecific PCR assays have been used. 18 Although the ATI and the hemagglutinin (HA) gene-based PCRs developed for the diagnosis and differentiation of OPXVs 2,15,18,20 have been effective, these assays may not be suitable for rapid (because the length of PCR products is long), routine, specific diagnosis of camelpox infections and their simultaneous differentiation. A specific diagnostic PCR for the detection of CMLV has not yet been reported.…”

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confidence: 99%

“…7,15 Second, to identify a target nucleotide sequence in the CMLV genome for the development of a novel CMLV-specific PCR, the sequencing of a partial fragment of the C18L gene was performed based on the authors' earlier observations. Conserved signature sequences were sought in the host range ankyrin repeat protein coding C18L gene of BPXV, and sequence analysis was subsequently used to devise a PCR strategy for the detection and differentiation of BPXV from other OPXVs.…”

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confidence: 99%

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A Polymerase Chain Reaction Strategy for the Diagnosis of Camelpox

et al. 2009

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Abstract. Camelpox is a contagious viral skin disease that is mostly seen in young camels. The disease is caused by the Camelpox virus (CMLV). In the present study, a polymerase chain reaction (PCR) assay based on the C18L gene (encoding ankyrin repeat protein) and a duplex PCR based on the C18L and DNA polymerase (DNA pol ) genes were developed. The former assay yields a specific amplicon of 243 bp of the C18L gene, whereas the duplex PCR yields 243-and 96-bp products of the C18L and DNA pol genes, respectively, in CMLV, and only a 96-bp product of the DNA pol gene in other orthopoxviruses. The limit of detection was as low as 0.4 ng of viral DNA. Both PCR assays were employed successfully for the direct detection and differentiation of CMLV from other orthopoxviruses, capripoxviruses, and parapoxviruses in both cell culture samples and clinical material. Furthermore, a highly sensitive SYBR Green dye-based, realtime PCR was optimized for quantitation of CMLV DNA. In the standard curve of the quantitative assay, the melting temperature of the specific amplicon at 77.6uC with peak measured fluorescence in dissociation plot was observed with an efficiency of 102%. To the authors' knowledge, this is the first report to describe a C18L gene-based PCR for specific diagnosis of camelpox infection.

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“…7). In most strains of VACV as well as in VARV, camelpox virus, and monkeypox virus the ATI protein is truncated as a result of frameshift mutations (2,26). This gives rise to several ORFs whose products include, in some strains, a highly immunogenic truncated product that is unable to form typical ATIs but that cross-reacts with antiserum raised against the 160-kDa protein (29).…”

Section: Mab Isolationmentioning

confidence: 99%

Identification and Characterization of Three Immunodominant Structural Proteins of Fowlpox Virus

Boulanger

Green

Jones

et al. 2002

J Virol

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Genes encoding fowlpox virus (FWPVAvipoxviruses, which replicate only in avian cells, have been developed as safe recombinant vectors for human vaccination (23,30). In mammalian cells, avipoxvirus replication is blocked but early gene expression occurs (45,46). Late gene expression occurs in some mammalian cell types infected by fowlpox virus (FWPV), the prototypic avipoxvirus, without productive virus replication, as morphogenesis appears to be blocked (42). FWPV has been much less intensively studied than vaccinia virus (VACV). It appears to form extracellular enveloped virus (EEV) by budding rather then wrapping (6,16 (27). The complete sequence of a pathogenic strain of FWPV (1) includes genes encoding homologs of at least 21 known VACV structural proteins, but FWPV appears to lack homologs of intracellular mature virus (IMV) proteins encoded by A27L and D8L as well as an intracellular enveloped virus protein encoded by A36R and the EEV proteins encoded by B5R, A33R, and A56R (1).Although genes encoding FWPV structural proteins have been identified by sequence homology, very little is known about the proteins they encode. We produced monoclonal antibodies (MAbs) and polyclonal antibodies against FWPV proteins as markers to study FWPV morphogenesis and identified genes encoding three FWPV immunodominant structural proteins. MATERIALS AND METHODSVirus and cells. The origins, propagation, and purification of FWPV strains (FP9, HP1, and Poxine) and canarypox virus (CNPV) have been described previously (5).MAbs. BALB/c mice were immunized twice intraperitonally 3 weeks apart with a crude preparation of Poxine in Quil A, a saponin-derived adjuvant (Superfos, Vedbaek, Denmark). For the second fusion, mice were injected twice 4 weeks apart with 100 g of sucrose gradient-purified FP9 virus (5). Hybridoma supernatants were screened by enzyme-linked immunosorbent assay or by indirect immunofluorescence assay, with Poxine-infected or noninfected chicken embryo fibroblasts (CEFs) as the antigen. The 3D9 MAb directed against the 39-kDa protein was kindly provided by C. P. Czerny.Polyclonal sera. Chicken hyperimmune sera against FP9, Poxine, or CNPV were produced as described previously (5).Radioimmunoprecipitation. Infected (multiplicity of infection, 10) or uninfected CEFs were incubated in methionine-free medium containing 2% dialyzed fetal calf serum. Two hours later, the medium was replaced with fresh medium containing 1.85 MBq of [ 35 S]methionine/ml and incubated for 2 h. The medium was removed, and the cells were washed once with phosphate-buffered saline (PBS) and lysed in 2 ml of radioimmunoprecipitation assay (RIPA) buffer (2.5 mM Tris [pH 7.2], 0.15 M NaCl, 1% Triton X-100, 0.1% sodium dodecyl sulfate [SDS], 1% sodium deoxycholate). Lysates were centrifuged for 15 min (Eppendorf centrifuge), and supernatants were stored at Ϫ70°C.Ascitic fluid (10 l) or hybridoma supernatant (500 l) was incubated for 1 h at 4°C with 50 l of 10% protein A-Sepharose CL-4B (Pharmacia) in 0.1 M sodium phosphate buffer, pH 8.1. After...

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Characterization of the gene encoding the A-type inclusion protein of camelpox virus and sequence comparison with other orthopoxviruses

Cited by 39 publications

References 22 publications

Proteome Analysis of Plasmodium falciparum Extracellular Secretory Antigens at Asexual Blood Stages Reveals a Cohort of Proteins with Possible Roles in Immune Modulation and Signaling

Proteome Analysis of Plasmodium falciparum Extracellular Secretory Antigens at Asexual Blood Stages Reveals a Cohort of Proteins with Possible Roles in Immune Modulation and Signaling

A Polymerase Chain Reaction Strategy for the Diagnosis of Camelpox

Identification and Characterization of Three Immunodominant Structural Proteins of Fowlpox Virus

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