Biochemical and biophysical characterization of light and heavy density hepatitis A virus particles: Evidence HAV is an RNA virus

Bradley, Daniel W.; Fields, Howard A.; McCaustland, Karen A.; Cook, E. H.; Gravelle, Clifton R.; Maynard, James E.

doi:10.1002/jmv.1890020212

Cited by 27 publications

(8 citation statements)

References 24 publications

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“…38,57]. Dense HAV particles isolated from CsCl gradients have also been reported to sediment as mature virions when subsequently centrifuged over sucrose gradients [58], agreeing with data presented here. The RNA in HAV dense particles was found to be sensitive to RNase degradation, whilst that in typical HAV capsids was not, further supporting a more open structure of these particles.…”

Section: Discussionsupporting

confidence: 81%

Hepatitis A Virus Replication: An Intermediate in the Uncoating Process

Bishop¹

2000

Intervirology

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Dense, RNase-sensitive, VP2-containing, non-infectious hepatitis A virus (HAV) particles were found to be formed at early times after the infection of cultured cells. These particles formed with kinetics mirroring those reported for HAV uncoating. The kinetics of the formation of dense HAV particles corresponded to a decrease in detectable, mature input virions, as detected by RNA dot blot hybridization of CsCl density gradient fractions. The dense HAV particles did not appear to have altered sedimentation coefficients, and as the fate of small capsid protein VP4 is not yet known, these particles cannot yet be termed ‘A particles’ or ‘infectosomes’, as have the uncoating intermediates in some picornavirus-cell systems.

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Section: Discussionsupporting

confidence: 81%

Hepatitis A Virus Replication: An Intermediate in the Uncoating Process

Bishop¹

2000

Intervirology

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“…Provost and coworkers [4] were the first to postulate that HAV was an RNA virus, on the basis of acridine orange staining, partial inactivation of infectivity by reaction with RNase type A, and the intracytoplasmic location of virus particles in experimentally infected marmoset liver cells. Bradley et al [42] also provided in direct evidence that HAV was an RNA virus by demonstrating that, after treatment at an alkaline pH of 10-11, the virus was susceptible to RNase but not to DNase, as determined by altered sedimentation rates in sucrose gra dients.…”

Section: Biochemical Characterization O F Ha V (I) Polypeptide Composmentioning

confidence: 95%

“…Sedi mentation analysis of HAV by rate-zonal ultracentrifugation through sucrose has also produced conflicting findings. Bradley and co workers [42] reported that virus particles from human feces with buoyant densities of 1.34 and 1.45 g/cm3 both had sedimentation coeffi cients of 157S in neutral sucrose gradients. However, when analyzed on sucrose gradients containing 1.5 M NaCl, the 1.45 g/cm3 popula tion of HAV sedimented at 230S, while the particles banding at 1.34 g/cm3 retained a sedimentation coefficient of 157S.…”

Section: Characterization Of Hepatitis a Virusmentioning

confidence: 99%

Biophysical and Biochemical Characterization of Hepatitis A Virus

et al. 1982

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Biophysical and biochemical analysis of hepatitis A virus has shown it to be a 27- to 32-nm icosahedral particle with 32 capsomers. The mature virion has a buoyant density of 1.33–1.34 g/cm³, a sedimentation coefficient of 156–160S, and is composed of four polypeptides with molecular weights of 30,000–33,000 (VPl), 24,000–27,000 (VP2), 21,000–23,000 (VP3), and 7,000–14,000 (VP4). The genome of hepatitis A virus consists of a single piece of single-stranded RNA which sediments at 32–35S and has a buoyant density of 1.64 g/cm³. The molecular weight of RNA is 2.25 × 10⁶ when measured under nondenaturing conditions and 2.8 × 10⁶ when measured under fully denaturing conditions. The genome contains a 40–80 nucleotide sequence of polyadenylic acid and is capable of infecting cell cultures. These findings, together with the observation that the virion is stable at pH 3.0 and resistant to ether and a temperature of 60° for 1 h, indicate that hepatitis A virus should now be classified as an Entero-virus within the family Picornaviridae.

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“…Several investigators have detected the presence of HAV antigen within the cyto plasm of infected cells [3,6,8]. However, less information is available concerning the pre cise subcellular localization of HAV once it enters the hepatocyte.…”

mentioning

confidence: 99%

Localization of Hepatitis A Virus Antigen to Specific Subcellular Fractions of Hepatitis-A-Infected Chimpanzee Liver Cells

Nc¹,

Fb²,

Jl³

1984

Intervirology

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The localization of hepatitis A virus antigen to specific subcellular fractions of infected chimpanzee liver cells was studied by solid-phase radioimmunoassay following a mild subcellular fractionation procedure designed to separate various organelles from the cytosol and the nuclei. Most of the antigenic activity (93%) was evenly divided between the cytosol fraction and the microsomal suspension. Within the microsomal fraction, more than 75% of the detectable antigen was associated with the smooth endoplasmic reticulum. Less than 4% of the total antigenic activity was localized to the nucleus. These data provide additional evidence that replication of hepatitis A virus occurs primarily within the cytoplasm of the host cell in close association with cellular membranes, consistent with that observed for other members of the genus Enterovirus.

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Biochemical and biophysical characterization of light and heavy density hepatitis A virus particles: Evidence HAV is an RNA virus

Abstract: Heavy density HAV was also shown to be sensitive to low concentrations of RNase. The results of these biophysical and biochemical studies strongly support the notion HAV is an enterovirus.

Cited by 27 publications

References 24 publications

Hepatitis A Virus Replication: An Intermediate in the Uncoating Process

Hepatitis A Virus Replication: An Intermediate in the Uncoating Process

Biophysical and Biochemical Characterization of Hepatitis A Virus

Localization of Hepatitis A Virus Antigen to Specific Subcellular Fractions of Hepatitis-A-Infected Chimpanzee Liver Cells

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