Abstract
Bufavirus strain 1 (BuV1), a member of the Protoparvovirus genus of the Parvoviridae, was first isolated from fecal samples of children with acute diarrhea in Burkina Faso. Since this initial discovery, BuVs have been isolated in several countries, including Finland, the Netherlands, and Bhutan, in pediatric patients exhibiting similar symptoms. Towards their characterization, the structures of virus-like particles of BuV1, BuV2, and BuV3, the current known genotypes, have been determined by cryo-electron microscopy and image reconstruction to 2.84, 3.79, and 3.25 Å, respectively. The BuVs, 65–73% identical in amino acid sequence, conserve the major viral protein, VP2, structure and general capsid surface features of parvoviruses. These include a core β-barrel (βB-βI), α-helix A, and large surface loops inserted between these elements in VP2. The capsid contains depressions at the icosahedral 2-fold and around the 5-fold axes, and has three separated protrusions surrounding the 3-fold axes. Structure comparison among the BuVs and to available parvovirus structures revealed capsid surface variations and capsid 3-fold protrusions that depart from the single pinwheel arrangement of the animal protoparvoviruses. These structures provide a platform to begin the molecular characterization of these potentially pathogenic viruses.
Highlights
Advances in DNA sequencing technology have led to the discovery of several new members of the ssDNA Parvoviridae that infect humans, including human bocaviruses 1 to 4 (HBoV1-HBoV4), cutavirus (CuV), tusavirus (TuV), bufavirus 1 to 3 (BuV1-BuV3), and human parvovirus 4 (PARV4) [1,2,3,4,5,6,7].These viruses are grouped into different genera based on the DNA sequence of their non-structural (NS) proteins [8]
The Bufavirus strain 1 (BuV1), BuV2, and BuV3 VP2 genes were cloned into the pFastBac1 plasmid to produce a recombinant baculovirus that expresses virus-like particles (VLPs) using the Bac-to-Bac protocol according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA) [12]
The resulting clarified supernatant were subjected to a 20% (w/v) sucrose cushion to pellet the VLPs by ultracentrifugation at 45,000 rpm (Beckman 70-Ti) for 3 h at 4 ◦ C
Summary
Advances in DNA sequencing technology have led to the discovery of several new members of the ssDNA Parvoviridae that infect humans, including human bocaviruses 1 to 4 (HBoV1-HBoV4), cutavirus (CuV), tusavirus (TuV), bufavirus 1 to 3 (BuV1-BuV3), and human parvovirus 4 (PARV4) [1,2,3,4,5,6,7] These viruses are grouped into different genera based on the DNA sequence of their non-structural (NS) proteins [8]. BuV2 was discovered in Burkina Faso, while BuV3 has been observed in other countries, including Bhutan, China, Finland, Netherlands, Thailand, and Turkey [4,9,10,11] This includes detection in nasal swabs in a low percentage (0.1%) of pediatric samples in Finland [12]. The viral loads are low and the prevalence among diarrheic patients is low
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