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Published in: Virology Journal 1/2014

Open Access 01-12-2014 | Research

Complete genome analysis of a frog virus 3 (FV3) isolate and sequence comparison with isolates of differing levels of virulence

Authors: Elizabeth A Morrison, Shawn Garner, Pierre Echaubard, David Lesbarrères, Christopher J Kyle, Craig R Brunetti

Published in: Virology Journal | Issue 1/2014

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Abstract

Background

Frog virus 3 (FV3) is the type species of the genus Ranavirus, and in the past few decades, FV3 infections have resulted in considerable morbidity and mortality in a range of wild and cultivated amphibian species in the Americas, Europe, and Asia. The reasons for the pathogenicity of FV3 are not well understood.

Findings

We investigated three FV3 isolates designated SSME, wt-FV3, and aza-Cr, and reported that our wt-FV3 and aza-Cr strains showed similar levels of virulence, while SSME was the least virulent in an in vivo study with Lithiobates pipiens tadpoles. Using 454 GS-FLX sequencing technology, we sequenced SSME and compared it to the published wt-FV3 genome. SSME had multiple amino acid deletions in ORFs 49/50L, 65L, 66L, and 87L, which may explain its reduced virulence. We also investigated repeat regions and found that repeat copy number differed between isolates, with only one group of 3 isolates and 1 pair of isolates being identical at all 3 locations.

Conclusions

In this study we have shown that genetic variability is present between closely related FV3 isolates, both in terms of deletions/insertions, and even more so at select repeat locations. These genomic areas with deletions/insertions may represent regions that affect virulence, and therefore require investigation. Furthermore, we have identified repeat regions that may prove useful in future phylogeographical tracking and identification of ranaviral strains across different environmental regions.
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Literature
1.
go back to reference Berger L, Speare R, Daszak P, Green DE, Cunningham AA, Goggin CL, Slocombe R, Ragan MA, Hyatt AD, McDonald KR, Hines HB, Lips KR, Marantelli G, Parkes H: Chytridiomycosis causes amphibian mortality associated with population declines in the rain forests of Australia and Central America. Proc Natl Acad Sci U S A 1998, 95: 9031-9036.PubMedPubMedCentralCrossRef Berger L, Speare R, Daszak P, Green DE, Cunningham AA, Goggin CL, Slocombe R, Ragan MA, Hyatt AD, McDonald KR, Hines HB, Lips KR, Marantelli G, Parkes H: Chytridiomycosis causes amphibian mortality associated with population declines in the rain forests of Australia and Central America. Proc Natl Acad Sci U S A 1998, 95: 9031-9036.PubMedPubMedCentralCrossRef
2.
go back to reference Daszak P, Berger L, Cunninham AA, Hyatt AD, Green DE, Speare R: Emerging infectious diseases and amphibian population declines. Emerg Infect Dis 1999, 5: 735-748.PubMedPubMedCentralCrossRef Daszak P, Berger L, Cunninham AA, Hyatt AD, Green DE, Speare R: Emerging infectious diseases and amphibian population declines. Emerg Infect Dis 1999, 5: 735-748.PubMedPubMedCentralCrossRef
3.
go back to reference Tidona CA, Schnitzler P, Kehm R, Darai G: Is the major capsid protein of iridoviruses a suitable target for the study of viral evolution? Virus Genes 1998, 16: 59-66.PubMedCrossRef Tidona CA, Schnitzler P, Kehm R, Darai G: Is the major capsid protein of iridoviruses a suitable target for the study of viral evolution? Virus Genes 1998, 16: 59-66.PubMedCrossRef
4.
go back to reference King A, Adams M, Carstens E, Lefkowitz E: Family Iridoviridae, In: Virus Taxonomy: Ninth Report of the International Committee on Taxonomy of Viruses. San Diego: Academic Press; 2012. pp 193–207 King A, Adams M, Carstens E, Lefkowitz E: Family Iridoviridae, In: Virus Taxonomy: Ninth Report of the International Committee on Taxonomy of Viruses. San Diego: Academic Press; 2012. pp 193–207
5.
go back to reference Daszak P, Cunningham A, Hyatt H: Infectious disease and amphibian population declines. Divers Distrib 2003, 9: 141-150.CrossRef Daszak P, Cunningham A, Hyatt H: Infectious disease and amphibian population declines. Divers Distrib 2003, 9: 141-150.CrossRef
6.
go back to reference Lesbarrères D, Balseiro A, Brunner J, Chinchar VG, Duffus A, Kerby J, Miller DL, Robert J, Schock DM, Waltzek T, Gray MJ: Ranavirus: past present and future. Biol Lett 2012, 8: 481-483.PubMedPubMedCentralCrossRef Lesbarrères D, Balseiro A, Brunner J, Chinchar VG, Duffus A, Kerby J, Miller DL, Robert J, Schock DM, Waltzek T, Gray MJ: Ranavirus: past present and future. Biol Lett 2012, 8: 481-483.PubMedPubMedCentralCrossRef
7.
go back to reference Marsh IB, Whittington RJ, O’Rourke B, Hyatt AD, Chisholm O: Rapid differentiation of Australian, European and American ranaviruses based on variation in major capsid protein gene sequence. Mol Cell Probes 2002, 16: 137-151.PubMedCrossRef Marsh IB, Whittington RJ, O’Rourke B, Hyatt AD, Chisholm O: Rapid differentiation of Australian, European and American ranaviruses based on variation in major capsid protein gene sequence. Mol Cell Probes 2002, 16: 137-151.PubMedCrossRef
8.
go back to reference Stuart SN, Chanson JS, Cox NA, Young BE, Rodrigues AS, Fischman DL, Waller RW: Status and trends of amphibian declines and extinctions worldwide. Science 2004, 306: 1783-1786.PubMedCrossRef Stuart SN, Chanson JS, Cox NA, Young BE, Rodrigues AS, Fischman DL, Waller RW: Status and trends of amphibian declines and extinctions worldwide. Science 2004, 306: 1783-1786.PubMedCrossRef
9.
go back to reference Muths E, Gallant AL, Campbell Grant EH, Battaglin WA, Green DE, Staiger JS, Walls SC, Gunzburger MS, Kearney RF: The amphibian research and monitoring initiative (ARMI): 5-year report. US Geological Survey. Sci Invest Rep 2006, 5224: 77. Muths E, Gallant AL, Campbell Grant EH, Battaglin WA, Green DE, Staiger JS, Walls SC, Gunzburger MS, Kearney RF: The amphibian research and monitoring initiative (ARMI): 5-year report. US Geological Survey. Sci Invest Rep 2006, 5224: 77.
10.
go back to reference Green DE, Converse KA, Schrader AK: Epizootiology of 64 amphibian morbidity and mortality events in the USA, 1996–2001. Ann N Y Acad Sci 2002, 969: 323-339.PubMedCrossRef Green DE, Converse KA, Schrader AK: Epizootiology of 64 amphibian morbidity and mortality events in the USA, 1996–2001. Ann N Y Acad Sci 2002, 969: 323-339.PubMedCrossRef
11.
go back to reference Chinchar V, Ranavirus (family Iridoviridae): Emerging cold-blooded killers. Arch Virol 2002, 147: 447-470.PubMedCrossRef Chinchar V, Ranavirus (family Iridoviridae): Emerging cold-blooded killers. Arch Virol 2002, 147: 447-470.PubMedCrossRef
12.
go back to reference Tan W, Barkman T, Chinchar V, Essani K: Comparative genomic analyses of frog virus 3, type species of the genus Ranavirus (family Iridoviridae). Virology 2004, 323: 70-84.PubMedCrossRef Tan W, Barkman T, Chinchar V, Essani K: Comparative genomic analyses of frog virus 3, type species of the genus Ranavirus (family Iridoviridae). Virology 2004, 323: 70-84.PubMedCrossRef
13.
go back to reference Chinchar VG, Bryan L, Wang J, Long S, Chinchar GD: Introduction of apoptosis in frog virus 3-infected cells. Virology 2003, 306: 303-312.PubMedCrossRef Chinchar VG, Bryan L, Wang J, Long S, Chinchar GD: Introduction of apoptosis in frog virus 3-infected cells. Virology 2003, 306: 303-312.PubMedCrossRef
14.
go back to reference Williams T, Barbosa-Solomieu V, Chinchar GD: A decade of advances in iridovirus research. In Advances in virus research, Volume 65. Edited by: Maramorosch K, Shatkin A. New York: Academic Press; 2005:173-248.CrossRef Williams T, Barbosa-Solomieu V, Chinchar GD: A decade of advances in iridovirus research. In Advances in virus research, Volume 65. Edited by: Maramorosch K, Shatkin A. New York: Academic Press; 2005:173-248.CrossRef
15.
go back to reference Cunningham AA, Langton TE, Bennett PM, Lewin JF, Drury SE, Gough RE, Macgregor SK: Pathological and microbiological findings from incidents of unusual mortality of the common frog ( Rana temporaria ). Philos Trans R Soc Lond B 1996, 351: 1539-1557.CrossRef Cunningham AA, Langton TE, Bennett PM, Lewin JF, Drury SE, Gough RE, Macgregor SK: Pathological and microbiological findings from incidents of unusual mortality of the common frog ( Rana temporaria ). Philos Trans R Soc Lond B 1996, 351: 1539-1557.CrossRef
16.
go back to reference Kidd-Ljunggren K, Miyakawa Y, Kidd A: Genetic variability in hepatitis B viruses. J Gen Virol 2002, 83: 1267-1280.PubMedCrossRef Kidd-Ljunggren K, Miyakawa Y, Kidd A: Genetic variability in hepatitis B viruses. J Gen Virol 2002, 83: 1267-1280.PubMedCrossRef
17.
go back to reference Stuyver L, De Gendt S, Van Geyt C, Zoulim F, Fried M, Schinazi RF, Rossau R: A new genotype of hepatitis B virus: complete genome and phyogenetic relatedness. J Gen Virol 2000, 81: 67-74.PubMedCrossRef Stuyver L, De Gendt S, Van Geyt C, Zoulim F, Fried M, Schinazi RF, Rossau R: A new genotype of hepatitis B virus: complete genome and phyogenetic relatedness. J Gen Virol 2000, 81: 67-74.PubMedCrossRef
18.
go back to reference Wang J, Hoper D, Beer M, Osterrieder N: Complete genome sequence of virulent duck enteritis virus (DEV) strain 2085 and comparison with genome sequences of virulent and attenuated DEV strains. Virus Res 2011, 160: 316-325.PubMedCrossRef Wang J, Hoper D, Beer M, Osterrieder N: Complete genome sequence of virulent duck enteritis virus (DEV) strain 2085 and comparison with genome sequences of virulent and attenuated DEV strains. Virus Res 2011, 160: 316-325.PubMedCrossRef
19.
20.
go back to reference Kass D, Batzer M: Genome organization/human. Life Sciences: Encyclopedia of; 2001:1-8. Kass D, Batzer M: Genome organization/human. Life Sciences: Encyclopedia of; 2001:1-8.
21.
go back to reference Chinchar V, Granoff A: Temperature-sensitive mutants of frog virus 3: biochemical and genetic characterization. J Virol 1986, 58: 192-202.PubMedPubMedCentral Chinchar V, Granoff A: Temperature-sensitive mutants of frog virus 3: biochemical and genetic characterization. J Virol 1986, 58: 192-202.PubMedPubMedCentral
22.
go back to reference Essani K, Goorha R, Granoff A: Mutation in a DNA-binding protein reveals an association between DNA-methyltransferase activity and a 26,000-Da polypeptide in frog virus 3-infected cells. Virology 1987, 161: 211-217.PubMedCrossRef Essani K, Goorha R, Granoff A: Mutation in a DNA-binding protein reveals an association between DNA-methyltransferase activity and a 26,000-Da polypeptide in frog virus 3-infected cells. Virology 1987, 161: 211-217.PubMedCrossRef
23.
go back to reference Lei X, Ou T, Zhu R, Zhang Q: Sequencing and analysis of the complete genome of Rana grylio virus (RGV). Arch Virol 2012, 157: 1559-1564.PubMedCrossRef Lei X, Ou T, Zhu R, Zhang Q: Sequencing and analysis of the complete genome of Rana grylio virus (RGV). Arch Virol 2012, 157: 1559-1564.PubMedCrossRef
24.
go back to reference Hyatt AD, Gould A, Zupanovic Z, Cunningham AA, Hengstberger S, Whittington RJ, Kattenbelt J, Coupar BE: Comparative studies of piscine and amphibian iridoviruses. Arch Virol 2000, 145: 301-331.PubMedCrossRef Hyatt AD, Gould A, Zupanovic Z, Cunningham AA, Hengstberger S, Whittington RJ, Kattenbelt J, Coupar BE: Comparative studies of piscine and amphibian iridoviruses. Arch Virol 2000, 145: 301-331.PubMedCrossRef
25.
go back to reference He J, Lu L, Deng M, He H, Weng S, Wang XH, Zhou SY, Long QX, Wang XZ, Chan SM: Sequence analysis of the complete genome of an iridovirus isolated from the tiger frog. Virology 2002, 292: 185-197.PubMedCrossRef He J, Lu L, Deng M, He H, Weng S, Wang XH, Zhou SY, Long QX, Wang XZ, Chan SM: Sequence analysis of the complete genome of an iridovirus isolated from the tiger frog. Virology 2002, 292: 185-197.PubMedCrossRef
26.
go back to reference Granoff A, Came P, Breeze D: Viruses and renal carcinoma of Rana pipiens. Virology 1966, 29: 133-148.PubMedCrossRef Granoff A, Came P, Breeze D: Viruses and renal carcinoma of Rana pipiens. Virology 1966, 29: 133-148.PubMedCrossRef
27.
go back to reference Puri B, Nelson WM, Henchal EA, Hoke CH, Eckels KH, Dubois DR, Porter KR, Hayes CG: Molecular analysis of dengue virus attenuation after serial passage in primary dog kidney cells. J Gen Virol 1997, 78: 2287-2291.PubMedCrossRef Puri B, Nelson WM, Henchal EA, Hoke CH, Eckels KH, Dubois DR, Porter KR, Hayes CG: Molecular analysis of dengue virus attenuation after serial passage in primary dog kidney cells. J Gen Virol 1997, 78: 2287-2291.PubMedCrossRef
28.
go back to reference Huang Y, Huang X, Liu H, Gong J, Ouyang Z, Cui H, Cao J, Zhao Y, Wang X, Jiang Y, Qin Q: Complete sequence determination of a novel reptile iridovirus isolated from soft-shelled turtle and evolutionary analysis of Iridoviridae. BMC Genomics 2009, 10: 224.PubMedPubMedCentralCrossRef Huang Y, Huang X, Liu H, Gong J, Ouyang Z, Cui H, Cao J, Zhao Y, Wang X, Jiang Y, Qin Q: Complete sequence determination of a novel reptile iridovirus isolated from soft-shelled turtle and evolutionary analysis of Iridoviridae. BMC Genomics 2009, 10: 224.PubMedPubMedCentralCrossRef
29.
go back to reference Boyle K, Arps L, Traktman P: Biochemical and genetic analysis of the vaccinia virus D5 protein. Multimerization-dependent APTase activity is required to support viral DNA replication. J Virol 2007, 81: 844-89.PubMedPubMedCentralCrossRef Boyle K, Arps L, Traktman P: Biochemical and genetic analysis of the vaccinia virus D5 protein. Multimerization-dependent APTase activity is required to support viral DNA replication. J Virol 2007, 81: 844-89.PubMedPubMedCentralCrossRef
30.
go back to reference Striepen B, Soldati D, Garcia-Reguet N, Dubremetz JF, Roos D: Targeting of soluble proteins to the rhoptries and micronemes in Toxoplasma gondii . Mol Biochem Parasitol 2001, 113: 45-53.PubMedCrossRef Striepen B, Soldati D, Garcia-Reguet N, Dubremetz JF, Roos D: Targeting of soluble proteins to the rhoptries and micronemes in Toxoplasma gondii . Mol Biochem Parasitol 2001, 113: 45-53.PubMedCrossRef
31.
go back to reference Jancovich JK, Davidson EW, Parameswaran N, Mao J, Chinchar VG, Collins JP, Jacobs BL, Storfer A: Evidence for emergence of an amphibian iridoviral disease because of human-enhanced spread. Mol Ecol 2005, 14: 213-224.PubMedCrossRef Jancovich JK, Davidson EW, Parameswaran N, Mao J, Chinchar VG, Collins JP, Jacobs BL, Storfer A: Evidence for emergence of an amphibian iridoviral disease because of human-enhanced spread. Mol Ecol 2005, 14: 213-224.PubMedCrossRef
32.
go back to reference Gray MJ, Miller DL, Hoverman JT: Ecology and pathology of amphibian ranaviruses. Dis Aquat Organ 2009, 87: 243-266.PubMedCrossRef Gray MJ, Miller DL, Hoverman JT: Ecology and pathology of amphibian ranaviruses. Dis Aquat Organ 2009, 87: 243-266.PubMedCrossRef
33.
go back to reference Mao J, Hedrick RP, Chinchar VG: Molecular characterization, sequence analysis, and taxonomic position of newly isolated fish iridoviruses. Virology 1997, 229: 212-220.PubMedCrossRef Mao J, Hedrick RP, Chinchar VG: Molecular characterization, sequence analysis, and taxonomic position of newly isolated fish iridoviruses. Virology 1997, 229: 212-220.PubMedCrossRef
34.
go back to reference Davis C, Field D, Metzgar D, Saiz R, Morin P: Numerous length polymorphisms at short tandem repeats in human cytomegalovirus. J Virol 1999, 73: 6265-6270.PubMedPubMedCentral Davis C, Field D, Metzgar D, Saiz R, Morin P: Numerous length polymorphisms at short tandem repeats in human cytomegalovirus. J Virol 1999, 73: 6265-6270.PubMedPubMedCentral
35.
go back to reference Barr JN, Whelan SPJ, Wertz GW: cis -acting signals involved in termination of vesicular stomatitis mRNA synthesis include the conserved AUAC and the U7 signal for polyadenylation. J Virol 1997, 71: 8718-8725.PubMedPubMedCentral Barr JN, Whelan SPJ, Wertz GW: cis -acting signals involved in termination of vesicular stomatitis mRNA synthesis include the conserved AUAC and the U7 signal for polyadenylation. J Virol 1997, 71: 8718-8725.PubMedPubMedCentral
36.
go back to reference Yamada N, Tanihara K, Takada A, Yorihuzi T, Tsutsumi M, Shimomura H, Tsuji T, Date T: Genetic organization and diversity of the 3′ non-coding region of the hepatitis C virus genome. Virology 1996, 223: 255-261.PubMedCrossRef Yamada N, Tanihara K, Takada A, Yorihuzi T, Tsutsumi M, Shimomura H, Tsuji T, Date T: Genetic organization and diversity of the 3′ non-coding region of the hepatitis C virus genome. Virology 1996, 223: 255-261.PubMedCrossRef
37.
go back to reference Gosner KL: A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 1960, 16: 183-190. Gosner KL: A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 1960, 16: 183-190.
38.
go back to reference Echaubard P, Little K, Pauli B, Lesbarrères D: Context-dependent effects of ranaviral infection on Northern Leopard frog life history traits. PLoS One 2010, 5: e13723.PubMedPubMedCentralCrossRef Echaubard P, Little K, Pauli B, Lesbarrères D: Context-dependent effects of ranaviral infection on Northern Leopard frog life history traits. PLoS One 2010, 5: e13723.PubMedPubMedCentralCrossRef
39.
go back to reference Gehan EA: A generalized wilcoxon test for comparing arbitrarily singly-censored samples. Biometrika 1965, 52: 203-223.PubMedCrossRef Gehan EA: A generalized wilcoxon test for comparing arbitrarily singly-censored samples. Biometrika 1965, 52: 203-223.PubMedCrossRef
40.
go back to reference Leung KM, Elashoff RM, Afifi AA: Censoring issues in survival analysis. Annu Rev Public Health 1997, 18: 83-104.PubMedCrossRef Leung KM, Elashoff RM, Afifi AA: Censoring issues in survival analysis. Annu Rev Public Health 1997, 18: 83-104.PubMedCrossRef
Metadata
Title
Complete genome analysis of a frog virus 3 (FV3) isolate and sequence comparison with isolates of differing levels of virulence
Authors
Elizabeth A Morrison
Shawn Garner
Pierre Echaubard
David Lesbarrères
Christopher J Kyle
Craig R Brunetti
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2014
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/1743-422X-11-46

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