Biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus

Bauer, Hans-Joachim; Schneider, Richard F.; Gelderblom, Hans R.; Friehmelt, V.; Monreal, G

doi:10.1007/bf01310955

Archives of Virology

1991

DOI: 10.1007/bf01310955

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Biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus

Hans-Joachim Bauer

Richard F. Schneider

Hans R. Gelderblom

et al.

Abstract: Two infectious components with buoyant densities of 1.40 g/cm3 and 1.45 g/cm3, designated as major (1.40) and minor (1.45) component, were detected by banding avian adeno-associated virus (AAAV) isopycnically in CsCl. In metrizamide, however, infectious AAAV banded only as a single peak at a density of 1.32 g/cm3. Biological as well as physicochemical properties of the two AAAV components recovered from CsCl density gradient were described. Concerning the minor (1.45) component, three experimental findings may… Show more

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“…Physicochemical studies revealed that the capsid of AAAV consists of three VP proteins similar to those of other AAVs. The buoyant density of AAAV in CsCl gradients (1.39 to 1.44 g/cm 3 ) is similar to what has been reported for all AAVs (6,30,68). One study (30) also provided a limited restriction endonuclease map of AAAV.…”

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confidence: 69%

Cloning of an Avian Adeno-Associated Virus (AAAV) and Generation of Recombinant AAAV Particles

Bossis

Chiorini

2003

J Virol

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Recent studies have proposed that adeno-associated viruses (AAVs) are not evolutionarily linked to other mammalian autonomous parvoviruses but are more closely linked to the autonomous parvoviruses of birds. To better understand the relationship between primate and avian AAVs (AAAVs), we cloned and sequenced the genome of an AAAV (ATCC VR-865) and generated recombinant AAAV particles. The genome of AAAV is 4,694 nucleotides in length and has organization similar to that of other AAVs. The entire genome of AAAV displays 56 to 65% identity at the nucleotide level with the other known AAVs. The AAAV genome has inverted terminal repeats of 142 nucleotides, with the first 122 forming the characteristic T-shaped palindromic structure. The putative Rep-binding element consists of a tandem (GAGY) 4 repeat, and the putative terminal resolution site (trs), CCGGT/CG, contains a single nucleotide substitution relative to the AAV 2 trs. The Rep open reading frame of AAAV displays 50 to 54% identity at the amino acid level with the other AAVs, with most of the diversity clustered at the carboxyl and amino termini. Comparison of the capsid proteins of AAAV and the primate dependoviruses indicate that divergent regions are localized to surface-exposed loops. Despite these sequence differences, we were able to produce recombinant AAAV particles carrying a lacZ reporter gene by cotransfection in 293T cells and were able to examine transduction efficiency in both chicken primary cells and several cell lines. Our findings indicate that AAAV is the most divergent AAV described to date but maintains all the characteristics unique to the genera of dependovirus.Adeno-associated viruses (AAVs) are small, nonpathogenic parvoviruses that require coinfection with a helper virus, such as adenovirus or herpesvirus, for productive infection (2). To date, eight AAV isolates (AAV types 1 to 8 [AAV1 to -8]) have been characterized and sequenced (2,4,19,20,25,32,51,56), with AAV2 having been the most extensively studied.AAV virions are approximately 20 to 25 nm in diameter and are composed of a mixture of assembled proteins (VPs) that encapsidate a linear ϳ4.7-kb single-stranded DNA (ssDNA) of plus or minus polarity (7, 43). The genome of AAVs is flanked by inverted terminal repeats (ITRs), which in the case of AAV 2 are 145 nucleotides in length. The ITR is organized as three interrupted palindromes that can fold in an energetically favored T-shaped hairpin structure, which can exist in two orientations, termed flip and flop (42). The ITRs serve as origin of replication and contain cis-acting elements required for rescue, integration, excision from cloning vectors, and packaging (41,42,49,58).The genetic map of the AAVs has been derived primarily from studies of AAV2 but is conserved in all serotypes (26,27,29,36,42,45,46,58,60,64). Two major open reading frames (rep and cap ORFs) and three transcriptional active promoters (P 5 , P 19 , and P 40 ) have been identified in the genome of AAV2. The P 5 and P 19 promoters encode the nonstructural rep...

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supporting

confidence: 69%

Cloning of an Avian Adeno-Associated Virus (AAAV) and Generation of Recombinant AAAV Particles

Bossis

Chiorini

2003

J Virol

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Molecular characterization of two strains of the avian adeno-associated virus (AAAV)

Heß

Paul

Kling

et al. 1995

Archives of Virology

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An avian adeno-associated virus (AAAV) was isolated after propagating a field isolate of the CELO virus (fowl adenovirus serotype 1 (FAV1)) in embryonated eggs. The isolated dependovirus was compared with the known AAAV obtained from the American Type Culture Collection (ATCC VR-865). The genomes were analysed by digestion with several restriction endonucleases. Although both DNAs have the same size, most restriction enzymes produced different restriction patterns. Double digests were used to construct for the first time restriction maps for avian dependoviruses. The two DNAs rendered different restriction maps in which the different restriction sites were mainly located in the middle and right part of the genomes. The effect of these differences on the structure proteins was shown by western blot analysis. In the immunoblot, the immunofluorescence and immunodiffusion test the two dependoviruses were serologically indistinguishable and therefore can be regarded as two different strains of the same virus. To differentiate between both strains we named the original one as AAAV VR-865 compared with the isolated AAAV DA-1.

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Biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus

Cited by 2 publications

References 24 publications

Cloning of an Avian Adeno-Associated Virus (AAAV) and Generation of Recombinant AAAV Particles

Cloning of an Avian Adeno-Associated Virus (AAAV) and Generation of Recombinant AAAV Particles

Molecular characterization of two strains of the avian adeno-associated virus (AAAV)

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