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Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
Published on: December 29, 2015
Characterization of two novel polyomaviruses of birds by using multiply primed rolling-circle amplification of their
Reimar Johne1, Walter Wittig, Daniel Fernández-de-Luco
1Institute for Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany. johne@vetmed.uni-leipzig.de
Abstract:
Polyomaviruses are small nonenveloped particles with a circular double-stranded genome, approximately 5 kbp in size. The mammalian polyomaviruses mainly cause persistent subclinical infections in their natural nonimmunocompromised hosts. In contrast, the polyomaviruses of birds--avian polyomavirus (APV) and goose hemorrhagic polyomavirus (GHPV)--are the primary agents of acute and chronic disease with high mortality rates in young birds. Screening of field samples of diseased birds by consensus PCR revealed the presence of two novel polyomaviruses in the liver of an Eurasian bullfinch (Pyrrhula pyrrhula griseiventris) and in the spleen of a Eurasian jackdaw (Corvus monedula), tentatively designated as finch polyomavirus (FPyV) and crow polyomavirus (CPyV), respectively. The genomes of the viruses were amplified by using multiply primed rolling-circle amplification and cloned. Analysis of the FPyV and CPyV genome sequences revealed a close relationship to APV and GHPV, indicating the existence of a distinct avian group among the polyomaviruses. The main characteristics of this group are (i) involvement in fatal disease, (ii) the existence of an additional open reading frame in the 5' region of the late mRNAs, and (iii) a different manner of DNA binding of the large tumor antigen compared to that of the mammalian polyomaviruses.
Insights
Two novel avian polyomaviruses, finch polyomavirus (FPyV) and crow polyomavirus (CPyV), were discovered in diseased birds. These viruses are closely related to other avian polyomaviruses and cause severe illness.
Area of Science:
- Virology
- Ornithology
- Molecular Biology
Background:
- Polyomaviruses are small, nonenveloped viruses with circular double-stranded DNA genomes.
- Mammalian polyomaviruses typically cause asymptomatic infections, while avian polyomaviruses like APV and GHPV cause severe disease in young birds.
Purpose of the Study:
- To identify and characterize novel polyomaviruses in diseased birds.
- To determine the phylogenetic relationship of newly identified avian polyomaviruses to known polyomaviruses.
Main Methods:
- Screening of field samples from diseased birds using consensus PCR.
- Genome amplification via multiply primed rolling-circle amplification and subsequent cloning.
- Sequence analysis of amplified viral genomes.
Main Results:
- Discovery of two novel polyomaviruses, finch polyomavirus (FPyV) and crow polyomavirus (CPyV), in an Eurasian bullfinch and a Eurasian jackdaw, respectively.
- Genomic analysis revealed FPyV and CPyV are closely related to avian polyomaviruses (APV, GHPV), forming a distinct avian group.
- Key characteristics of this avian group include association with fatal disease, an additional ORF in late mRNAs, and distinct large tumor antigen DNA binding.
Conclusions:
- Finch polyomavirus and crow polyomavirus represent a distinct group of avian polyomaviruses.
- This avian polyomavirus group is associated with severe, fatal diseases in birds.
- Genomic and functional differences distinguish avian polyomaviruses from their mammalian counterparts.
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