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Abstract

Koala retrovirus (KoRV) is unique amongst endogenous (inherited) retroviruses in that its incorporation to the host genome is still active, providing an opportunity to study what drives this fundamental process in vertebrate genome evolution. Animals in the southern part of the natural range of koalas were previously thought to be either virus-free or to have only exogenous variants of KoRV with low rates of KoRV-induced disease. In contrast, animals in the northern part of their range universally have both endogenous and exogenous KoRV with very high rates of KoRV-induced disease such as lymphoma. In this study we use a combination of sequencing technologies, Illumina RNA sequencing of ‘southern’ (south Australian) and ‘northern’ (SE QLD) koalas and CRISPR enrichment and nanopore sequencing of DNA of ‘southern’ (South Australian and Victorian animals) to retrieve full-length loci and intregration sites of KoRV variants. We demonstrate that koalas that tested negative to the KoRV gene qPCR, used to detect replication-competent KoRV, are not in fact KoRV-free but harbour defective, presumably endogenous, ‘RecKoRV’ variants that are not fixed between animals. This indicates that these populations have historically been exposed to KoRV and raises questions as to whether these variants have arisen by chance or whether they provide a protective effect from the infectious forms of KoRV. This latter explanation would offer the intriguing prospect of being able to monitor and selectively breed for disease resistance to protect the wild koala population from KoRV-induced disease.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2022-06-28
2024-04-18
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References

  1. Tarlinton RE. Differential and defective transcription of koala retrovirus indicate complexity of host and virus evolution. Figshare 2022 [View Article]
    [Google Scholar]
  2. Woinarski J, Burbidge AA. Phascolarctos cinereus. the IUCN red list of threatened species 2016 e.T16892A21960344 (2016); 2016
  3. Environment and Communications Reference Committee The koala saving our national icon (senate printing unit). Parliament House Canberra 2011
    [Google Scholar]
  4. Polkinghorne A, Hanger J, Timms P. Recent advances in understanding the biology, epidemiology and control of chlamydial infections in koalas. Vet Microbiol 2013; 165:214–223 [View Article] [PubMed]
    [Google Scholar]
  5. Simmons GS, Young PR, Hanger JJ, Jones K, Clarke D et al. Prevalence of koala retrovirus in geographically diverse populations in Australia. Aust Vet J 2012; 90:404–409 [View Article] [PubMed]
    [Google Scholar]
  6. Neaves LE, Frankham GJ, Dennison S, FitzGibbon S, Flannagan C et al. Phylogeography of the Koala, (Phascolarctos cinereus), and Harmonising Data to Inform Conservation. PLoS One 2016; 11:e0162207 [View Article] [PubMed]
    [Google Scholar]
  7. Tarlinton RE, Fabijan J, Hemmatzadeh F, Meers J, Owen H et al. Transcriptomic and genomic variants between koala populations reveals underlying genetic components to disorders in a bottlenecked population. Conserv Genet 2021; 22:329–340 [View Article]
    [Google Scholar]
  8. Robinson A. C. in The koala: Proceedings of the Taronga Symposium on Koala Biology, Management and Medicine Sydney: Zoological Parks Board; pp 132–143
    [Google Scholar]
  9. lindsay HA. Re-establishing the koala in SA. Wild Life 1950; 12:257–262
    [Google Scholar]
  10. Hayward A, Grabherr M, Jern P. Broad-scale phylogenomics provides insights into retrovirus-host evolution. Proc Natl Acad Sci U S A 2013; 110:20146–20151 [View Article] [PubMed]
    [Google Scholar]
  11. Manghera M, Douville RN. Endogenous retrovirus-K promoter: a landing strip for inflammatory transcription factors?. Retrovirology 2013; 10:16 [View Article] [PubMed]
    [Google Scholar]
  12. Chuong EB, Rumi MAK, Soares MJ, Baker JC. Endogenous retroviruses function as species-specific enhancer elements in the placenta. Nat Genet 2013; 45:325–329 [View Article] [PubMed]
    [Google Scholar]
  13. Chen C-P, Chen L-F, Yang S-R, Chen C-Y, Ko C-C et al. Functional characterization of the human placental fusogenic membrane protein syncytin 2. Biol Reprod 2008; 79:815–823 [View Article] [PubMed]
    [Google Scholar]
  14. Dupressoir A, Marceau G, Vernochet C, Bénit L, Kanellopoulos C et al. Syncytin-A and syncytin-B, two fusogenic placenta-specific murine envelope genes of retroviral origin conserved in Muridae. Proc Natl Acad Sci U S A 2005; 102:725–730 [View Article] [PubMed]
    [Google Scholar]
  15. Grow EJ, Flynn RA, Chavez SL, Bayless NL, Wossidlo M et al. Intrinsic retroviral reactivation in human preimplantation embryos and pluripotent cells. Nature 2015; 522:221–225 [View Article] [PubMed]
    [Google Scholar]
  16. Watanabe S, Kawamura M, Odahara Y, Anai Y, Ochi H et al. Phylogenetic and structural diversity in the feline leukemia virus env gene. PLoS One 2013; 8:e61009 [View Article] [PubMed]
    [Google Scholar]
  17. Evans LH, Alamgir ASM, Owens N, Weber N, Virtaneva K et al. Mobilization of endogenous retroviruses in mice after infection with an exogenous retrovirus. J Virol 2009; 83:2429–2435 [View Article]
    [Google Scholar]
  18. Hanger JJ, Bromham LD, McKee JJ, O’Brien TM, Robinson WF. The nucleotide sequence of Koala (Phascolarctos cinereus) Retrovirus: a Novel Type C Endogenous Virus Related to Gibbon Ape Leukemia Virus. J Virol 2000; 74:4264–4272 [View Article] [PubMed]
    [Google Scholar]
  19. Tarlinton RE, Meers J, Young PR. Retroviral invasion of the koala genome. Nature 2006; 442:79–81 [View Article]
    [Google Scholar]
  20. Sarker N, Fabijan J, Owen H, Seddon J, Simmons G et al. Koala retrovirus viral load and disease burden in distinct northern and southern koala populations. Sci Rep 2020; 10:263 [View Article]
    [Google Scholar]
  21. Maher IE, Higgins DP. Altered immune cytokine expression associated with KoRV B infection and season in captive koalas. PLoS One 2016; 11:e0163780 [View Article]
    [Google Scholar]
  22. McEwen GK, Alquezar-Planas DE, Dayaram A, Gillett A, Tarlinton R et al. Retroviral integrations contribute to elevated host cancer rates during germline invasion. Nat Commun 2021; 12:1316 [View Article] [PubMed]
    [Google Scholar]
  23. Quigley BL, Phillips S, Olagoke O, Robbins A, Hanger J et al. Changes in endogenous and exogenous koala retrovirus subtype expression over time reflect koala health outcomes. J Virol 2019; 93:e00849-19 [View Article] [PubMed]
    [Google Scholar]
  24. Waugh CA, Hanger J, Loader J, King A, Hobbs M et al. Infection with koala retrovirus subgroup B (KoRV-B), but not KoRV-A, is associated with chlamydial disease in free-ranging koalas (Phascolarctos cinereus). Sci Rep 2017; 7:134 [View Article] [PubMed]
    [Google Scholar]
  25. Legione AR, Patterson JLS, Whiteley P, Firestone SM, Curnick M et al. Koala retrovirus genotyping analyses reveal A low prevalence of KoRV-A in Victorian koalas and an association with clinical disease. J Med Microbiol 2017; 66:236–244 [View Article] [PubMed]
    [Google Scholar]
  26. Tarlinton R, Meers J, Hanger J, Young P. Real-time reverse transcriptase PCR for the endogenous koala retrovirus reveals an association between plasma viral load and neoplastic disease in koalas. J Gen Virol 2005; 86:783–787 [View Article] [PubMed]
    [Google Scholar]
  27. Fabijan J, Miller D, Olagoke O, Woolford L, Boardman W et al. Prevalence and clinical significance of koala retrovirus in two South Australian koala (Phascolarctos cinereus) populations. J Med Microbiol 2019; 68:1072–1080 [View Article] [PubMed]
    [Google Scholar]
  28. Quigley BL, Melzer A, Ellis W, Tzipori G, Nilsson K et al. Koala retrovirus in northern australia shows a mixture of stable endogenization and exogenous lineage diversification within fragmented koala populations. J Virol 2021JVI.02084-20 [View Article] [PubMed]
    [Google Scholar]
  29. Chappell KJ, Brealey JC, Amarilla AA, Watterson D, Hulse L et al. Phylogenetic diversity of koala retrovirus within a wild koala population. J Virol 2017; 91:e01820-16 [View Article] [PubMed]
    [Google Scholar]
  30. Cui P, Löber U, Alquezar-Planas DE, Ishida Y, Courtiol A et al. Comprehensive profiling of retroviral integration sites using target enrichment methods from historical koala samples without an assembled reference genome. PeerJ 2016; 4:e1847 [View Article] [PubMed]
    [Google Scholar]
  31. Sarker N, Fabijan J, Seddon J, Tarlinton R, Owen H et al. Genetic diversity of Koala retrovirus env gene subtypes: insights into northern and southern koala populations. J Gen Virol 2019; 100:1328–1339 [View Article] [PubMed]
    [Google Scholar]
  32. Sarker N, Fabijan J, Owen H, Seddon J, Simmons G et al. Koala retrovirus viral load and disease burden in distinct northern and southern koala populations. Sci Rep 2020; 10:263 [View Article] [PubMed]
    [Google Scholar]
  33. Hobbs M, King A, Salinas R, Chen Z, Tsangaras K et al. Long-read genome sequence assembly provides insight into ongoing retroviral invasion of the koala germline. Sci Rep 2017; 7:15838 [View Article] [PubMed]
    [Google Scholar]
  34. Quigley BL, Wedrowicz F, Hogan F, Timms P. Phylogenetic and geographical analysis of a retrovirus during the early stages of endogenous adaptation and exogenous spread in a new host. Mol Ecol 2021; 30:2626–2640 [View Article] [PubMed]
    [Google Scholar]
  35. Xu W, Stadler CK, Gorman K, Jensen N, Kim D et al. An exogenous retrovirus isolated from koalas with malignant neoplasias in a US zoo. Proc Natl Acad Sci U S A 2013; 110:11547–11552 [View Article] [PubMed]
    [Google Scholar]
  36. Quigley BL, Ong VA, Hanger J, Timms P. Molecular dynamics and mode of transmission of koala retrovirus as it invades and spreads through a wild queensland koala population. J Virol 2018; 92:e01871-17 [View Article] [PubMed]
    [Google Scholar]
  37. Zheng H, Pan Y, Tang S, Pye GW, Stadler CK et al. Koala retrovirus diversity, transmissibility, and disease associations. Retrovirology 2020; 17:34 [View Article] [PubMed]
    [Google Scholar]
  38. Hashem MA, Maetani F, Kayesh MEH, Eiei T, Mochizuki K et al. Transmission of koala retrovirus from parent koalas to a joey in a Japanese zoo. J Virol 2020; 94:e00019-20 [View Article] [PubMed]
    [Google Scholar]
  39. Xu W, Gorman K, Santiago JC, Kluska K, Eiden MV. Genetic diversity of koala retroviral envelopes. Viruses 2015; 7:1258–1270 [View Article] [PubMed]
    [Google Scholar]
  40. Shimode S, Nakagawa S, Yoshikawa R, Shojima T, Miyazawa T. Heterogeneity of koala retrovirus isolates. FEBS Lett 2014; 588:41–46 [View Article] [PubMed]
    [Google Scholar]
  41. Xu W, Eiden MV. Koala retroviruses: evolution and disease dynamics. Annu Rev Virol 2015; 2:119–134 [View Article] [PubMed]
    [Google Scholar]
  42. Shojima T, Yoshikawa R, Hoshino S, Shimode S, Nakagawa S et al. Identification of a novel subgroup of Koala retrovirus from Koalas in Japanese zoos. J Virol 2013; 87:9943–9948 [View Article] [PubMed]
    [Google Scholar]
  43. Yu T, Koppetsch BS, Pagliarani S, Johnston S, Silverstein NJ et al. The piRNA response to retroviral invasion of the koala genome. Cell 2019; 179:632–643 [View Article] [PubMed]
    [Google Scholar]
  44. Löber U, Hobbs M, Dayaram A, Tsangaras K, Jones K et al. Degradation and remobilization of endogenous retroviruses by recombination during the earliest stages of a germ-line invasion. Proc Natl Acad Sci U S A 2018; 115:8609–8614 [View Article] [PubMed]
    [Google Scholar]
  45. Martin R, Handasyde K. The Koala: Natural History, Conservation and Management Sydney: University of New South Wales Press; 1999
    [Google Scholar]
  46. Tarlinton R, Meers J, Hanger J, Young P. Real-time reverse transcriptase PCR for the endogenous koala retrovirus reveals an association between plasma viral load and neoplastic disease in koalas. J Gen Virol 2005; 86:783–787 [View Article] [PubMed]
    [Google Scholar]
  47. Martin RW. Age-specific fertility in three populations of the koala, phascolarctos cinereus goldfuss, in Victoria. Wildl Res 1981; 8:275 [View Article]
    [Google Scholar]
  48. Jiang H, Lei R, Ding SW, Zhu S. Skewer: a fast and accurate adapter trimmer for next-generation sequencing paired-end reads. BMC Bioinformatics 2014; 15:182 [View Article] [PubMed]
    [Google Scholar]
  49. Kim D, Langmead B, Salzberg SL. HISAT: a fast spliced aligner with low memory requirements. Nat Methods 2015; 12:357–360 [View Article] [PubMed]
    [Google Scholar]
  50. Johnson RN, O’Meally D, Chen Z, Etherington GJ, Ho SYW et al. Adaptation and conservation insights from the koala genome. Nat Genet 2018; 50:1102–1111 [View Article] [PubMed]
    [Google Scholar]
  51. Bray NL, Pimentel H, Melsted P, Pachter L. Near-optimal probabilistic RNA-seq quantification. Nat Biotechnol 2016; 34:525–527 [View Article] [PubMed]
    [Google Scholar]
  52. Li H. Minimap2: pairwise alignment for nucleotide sequences. Bioinformatics 2018; 34:3094–3100 [View Article] [PubMed]
    [Google Scholar]
  53. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J et al. The Sequence Alignment/Map format and SAMtools. Bioinformatics 2009; 25:2078–2079 [View Article] [PubMed]
    [Google Scholar]
  54. Hobbs M, Pavasovic A, King AG, Prentis PJ, Eldridge MDB et al. A transcriptome resource for the Koala (Phascolarctos cinereus): insights into koala retrovirus transcription and sequence diversity. BMC Genomics 2014; 15:786 [View Article] [PubMed]
    [Google Scholar]
  55. Kolmogorov M, Yuan J, Lin Y, Pevzner PA. Assembly of long, error-prone reads using repeat graphs. Nat Biotechnol 2019; 37:540–546 [View Article] [PubMed]
    [Google Scholar]
  56. Fabijan J, Sarker N, Speight N, Owen H, Meers J et al. Pathological findings in koala retrovirus-positive Koalas (Phascolarctos cinereus) from Northern and Southern Australia. J Comp Pathol 2020; 176:50–66 [View Article] [PubMed]
    [Google Scholar]
  57. Shojima T, Hoshino S, Abe M, Yasuda J, Shogen H et al. Construction and characterization of an infectious molecular clone of Koala retrovirus. J Virol 2013; 87:5081–5088 [View Article] [PubMed]
    [Google Scholar]
  58. Magiorkinis G, Gifford RJ, Katzourakis A, De Ranter J, Belshaw R. Env-less endogenous retroviruses are genomic superspreaders. Proc Natl Acad Sci U S A 2012; 109:7385–7390 [View Article] [PubMed]
    [Google Scholar]
  59. Rubin H. The early history of tumor virology: Rous, RIF, and RAV. Proc Natl Acad Sci U S A 2011; 108:14389–14396 [View Article] [PubMed]
    [Google Scholar]
  60. Nethe M, Berkhout B, van der Kuyl AC. Retroviral superinfection resistance. Retrovirology 2005; 2:52 [View Article] [PubMed]
    [Google Scholar]
  61. Spencer TE, Mura M, Gray CA, Griebel PJ, Palmarini M. Receptor usage and fetal expression of ovine endogenous betaretroviruses: implications for coevolution of endogenous and exogenous retroviruses. J Virol 2003; 77:749–753 [View Article] [PubMed]
    [Google Scholar]
  62. van der Kuyl AC. HIV infection and HERV expression: a review. Retrovirology 2012; 9:6 [View Article] [PubMed]
    [Google Scholar]
  63. Chiu ES, McDonald CA, VandeWoude S. Endogenous Feline Leukemia Virus (FeLV) siRNA transcription may interfere with exogenous FeLV Infection. J Virol 2021; 95:e0007021 [View Article] [PubMed]
    [Google Scholar]
  64. Chiu ES, VandeWoude S. Endogenous retroviruses drive resistance and promotion of exogenous retroviral homologs. Annu Rev Anim Biosci 2021; 9:225–248 [View Article] [PubMed]
    [Google Scholar]
  65. Chiu ES, VandeWoude S. Presence of endogenous viral elements negatively correlates with feline leukemia virus susceptibility in puma and domestic cat cells. J Virol 2020; 94:e01274-20 [View Article] [PubMed]
    [Google Scholar]
  66. Olagoke O, Miller D, Hemmatzadeh F, Stephenson T, Fabijan J et al. Induction of neutralizing antibody response against koala retrovirus (KoRV) and reduction in viral load in koalas following vaccination with recombinant KoRV envelope protein. NPJ Vaccines 2018; 3:30 [View Article] [PubMed]
    [Google Scholar]
  67. Olagoke O, Quigley BL, Hemmatzadeh F, Tzipori G, Timms P. Therapeutic vaccination of koalas harbouring endogenous koala retrovirus (KoRV) improves antibody responses and reduces circulating viral load. NPJ Vaccines 2020; 5:60 [View Article] [PubMed]
    [Google Scholar]
  68. Fabijan J, Caraguel C, Jelocnik M, Polkinghorne A, Boardman WSJ et al. Chlamydia pecorum prevalence in South Australian koala (Phascolarctos cinereus) populations: Identification and modelling of a population free from infection. Sci Rep 2019; 9:6261 [View Article] [PubMed]
    [Google Scholar]
  69. Speight KN, Haynes JI, Boardman W, Breed WG, Taggart DA et al. Plasma biochemistry and urinalysis variables of koalas (Phascolarctos cinereus) with and without oxalate nephrosis. Vet Clin Pathol 2014; 43:244–254 [View Article] [PubMed]
    [Google Scholar]
  70. Sullivan MJ, Petty NK, Beatson SA. Easyfig: a genome comparison visualizer. Bioinformatics 2011; 27:1009–1010 [View Article] [PubMed]
    [Google Scholar]
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