The Genome of Hepatitis B Virus

Robinson, William S.

doi:10.1146/annurev.mi.31.100177.002041

Cited by 103 publications

(46 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…From the analysis of the restriction fragments of HBV DNA and, particularly, from the existence of the Hae III AA' doublet, Robinson and coworkers (6,16) suggest that the HBV DNA population is heterogeneous. On heteroduplexes and HBV DNA homoduplexes, as exemplified in Fig.…”

Section: Discussionmentioning

confidence: 99%

Cloning in Escherichia coli and physical structure of hepatitis B virion DNA.

Charnay¹,

Pourcel²,

Louise³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

154

View full text Add to dashboard Cite

A restriction map of hepatitis B virion DNA was established after cloning of the whole viral genome in Escherichia coli. By use of EcoRI, Xho I, Bgl II, Xba I, BamHI, HincII, and Hae III endonucleases, a total of 28 restriction sites were mapped. The single-stranded region was localized on the restriction map and the 5' end of the short strand was mapped at a fixed position. Hepatitis B is a frequent and sometimes severe disease. In some areas of Asia and tropical Africa, 10% of the population carry the viral surface antigen (1). The Dane particle (2), found in the sera of patients, is considered to be the etiological agent (3). It contains a circular DNA molecule, hepatitis B virion (HBV) DNA, with a nick (or a short gap) and a single-stranded region (4-6). HBV DNA is the smallest known mammalian viral genome. These unique characteristics of the genome, the frequency of hepatitis B, and the likely relationship between this virus and primary liver carcinoma (7,8) justify an accurate investigation of the structure of the HBV genome.In this article we present a physical map of the HBV genome. Twenty eight restriction sites are located, and new information concerning the physical structure of HBV DNA is also established. In particular, the single-stranded region is localized on the restriction map. The position of the nick is discussed. MATERIALS AND METHODSDNA Preparations and Cloning. HBV were purified as described by Summers et al. (4) and their DNA was labeled with the endogenous DNA polymerase (6), with [32P]dTTP (22 Ci/mmol, 1 Ci = 3.7 X 1010 becquerels) as the radioactive precursor. After purification of the HBV DNA (4), it appeared from electron microscope observation that, under the action of the endogenous DNA polymerase, the single-stranded region of about 10% of the DNA molecules was totally repaired.The purified HBV DNA was used for cloning. The circular HBV genome was cleaved by EcoRI endonuclease at its unique restriction site (J. Summers, personal communication) and inserted in vitro into the DNA of bacteriophage Xgt WES. XB as described (9). After transfection of C600 recBC bacteria with the hybrid DNA, recombinant phage clones were purified three times. For each recombinant clone a first phage stock was made and, in order to avoid any genetic drift, all subsequent stocks were made from this initial sample. The cloned EcoRI DNA fragment will be referred to as Eco HBV DNA.After digestion of the DNA of a recombinant clone with EcoRI endonuclease, Eco HBV DNA was purified from the two vector arms by zone centrifugation in a 5-30% sucrose gradient.Enzymatic Digestions and Chemicals. Digestions by the following restriction nucleases (New England BioLabs), EcoRI, BamHI, Sma I,Xho I, Pst I, Xba I, Bgl II, Sst I, Sal I, Kpn I, HincII, and Hae III, were performed in the buffers recommended by New England BioLabs. Alkaline phosphatase (P-L Biochemicals) was used in 10 mM Tris-HCI, pH 8.5/10 mM MgCl2. The 5' ends were labeled by means of [y-32P]ATP (New England Nuclear, 3000 Ci/mmol) and polynucleoti...

show abstract

Section: Discussionmentioning

confidence: 99%

Cloning in Escherichia coli and physical structure of hepatitis B virion DNA.

Charnay¹,

Pourcel²,

Louise³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

154

View full text Add to dashboard Cite

show abstract

“…Structurally, hepatitis B virus (HBV) is similar to other lipidenveloped viruses in that it has an inner nucleocapsid surrounded by an outer lipoprotein envelope which has hepatitis B surface antigen (HBsAg)-reactive determinants (21). The intact core component possesses an immunologically distinct hepatitis B core antigen (HBcAg) reactivity which on further disruption reveals the third HBV-associated antigen, hepatitis B e antigen (21). Also present in the sera of infected individuals are 22-nm HBsAg particles, which have been shown by biochemical analysis to contain approximately 23% lipid by dry weight in addition to protein and carbohydrates (25).…”

mentioning

confidence: 99%

Effects of amphotericin B on hepatitis B virus

Kessler¹,

Dixon²,

Howard³

et al. 1981

Antimicrob Agents Chemother

View full text Add to dashboard Cite

We investigated the effects of amphotericin B (AmB) on the ultrastructure and biochemistry of hepatitis B virus (HBV) core component possesses an immunologically distinct hepatitis B core antigen (HBcAg) reactivity which on further disruption reveals the third HBV-associated antigen, hepatitis B e antigen (21). Also present in the sera of infected individuals are 22-nm HBsAg particles, which have been shown by biochemical analysis to contain approximately 23% lipid by dry weight in addition to protein and carbohydrates (25). Neutral lipids account for 22.3% of this fraction, with 59% of these being cholesterol and cholesterol esters (25). We predicted, therefore, that AmB would have an effect on the structural integrity of HBV particles, assuming the outer lipid membrane to be of approximately the same chemical composition as HBsAg particles. This study was therefore undertaken to determine the ultrastructural effects of AmB on HBV and the 22-nm form of HBsAg as determined by HBV deoxyribonucleic acid (DNA) polymerase activity, electron microscopy (EM), differential ultracentrifugation, and sucrose density centrifugation.MATERIALS AND METHODS HBV and HBsAg. Partially purified HBV and purified 22-nm HBsAg particles were obtained as previously described from a chimpanzee chronically infected with HBV (10, 22). Radioiodinated purified HBsAg particles were prepared as described by Skelly et al. (23). HBV-associated DNA polymerase activity was determined by the method of Howard (9)

show abstract

“…The virion (also known as the Dane particle) has a complex structure (1, 2) with hepatitis B surface antigen (HBsAg) on its outer lipid-containing envelope (1,2) and the distinct hepatitis B core antigen (HBcAg) on its internal core or nucleocapsid (2). Substantial evidence indicates that the HBsAg is specified by the viral genome (3). A group-specific determinant a found in all HBsAg preparations and two pairs of subtype determinants (d, y and w, r), which are for the most part mutually exclusive and thus behave as alleles, have been described (4,5).…”

mentioning

confidence: 99%

Restriction endonuclease cleavage map and location of unique features of the DNA of hepatitis B virus, subtype adw2.

Siddiqui¹,

Sattler²,

Robinson³

1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Hepatitis B virus (HBV) has several interesting features. The virion (also known as the Dane particle) has a complex structure (1, 2) with hepatitis B surface antigen (HBsAg) on its outer lipid-containing envelope (1, 2) and the distinct hepatitis B core antigen (HBcAg) on its internal core or nucleocapsid (2). Substantial evidence indicates that the HBsAg is specified by the viral genome (3). A group-specific determinant a found in all HBsAg preparations and two pairs of subtype determinants (d, y and w, r), which are for the most part mutually exclusive and thus behave as alleles, have been described (4, 5). Antigenic heterogeneity of the w determinant and additional determinants such as q and x or g have been found (6). Virus strains of the eight HBsAg subtypes aywi, ayw2, ayw3, ayw4, ayr, adw2, adw4, and adr as well as more complex and less common subtypes such as adyw and adywr ha've been identified (7). Both group-specific and type-specific HBsAg determinants appear to reside in the major 22,00dalton polypeptide, which can be isolated from purified HBsAg particles (8).Hepatitis B virions contain a circular DNA molecule (9) We have constructed a map of restriction endonuclease cleavage sites of HBV DNA (HBsAg subtype adw2) and have localized certain physical features of the DNA within this map, including the single-stranded region, the 5' end of the short strand, and the nick in the long strand. We have also found systematic differences in the restriction endonuclease cleavage patterns for DNA of HBV of HBsAg subtypes adw2, ayw3, and adrq+. MATERIALS AND METHODSPlasma rich in Dane particles was obtained by plasmaphoresis from patients persistently infected with HBV (HBsAg carriers). HBsAg subtyping was kindly performed by A. M. CouroucePauty. The detailed methods of Dane particle preparation and DNA extraction have been described (9). The single-stranded region of HBV DNA was always made double-stranded by an extensive virion DNA polymerase reaction in the presence of high deoxynucleoside triphosphate (dNTP) concentrations as described (12)

show abstract

The Genome of Hepatitis B Virus

Cited by 103 publications

References 30 publications

Cloning in Escherichia coli and physical structure of hepatitis B virion DNA.

Cloning in Escherichia coli and physical structure of hepatitis B virion DNA.

Effects of amphotericin B on hepatitis B virus

Restriction endonuclease cleavage map and location of unique features of the DNA of hepatitis B virus, subtype adw2.

Contact Info

Product

Resources

About