Adsorption and Penetration of Hepatitis B Virus in a Nonpermissive Cell Line

Qiao, Ming; Macnaughton, Thomas B.; Gowans, Eric J.

doi:10.1006/viro.1994.1301

Cited by 37 publications

(44 citation statements)

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“…Internalized viral DNA was detected according to a well established procedure (4,12,17,43,44) by incubating the cells first with the virus suspension for 2 h at 37°C and subsequently by treating the cells with trypsin. To prove that trypsinization removes bound HBV from the cell surface, control cells were incubated in parallel with virus suspension at 4°C, not allowing HBV penetration.…”

Section: Hbv-bp Is Responsible For Hbv Entry In Transfected Cells Andmentioning

confidence: 99%

“…They are the translational products of three overlapping open reading frames that start from different initiation codons localized at the 5Ј end of the preS1, preS2, and S regions of the env gene (2). The role of the preS1 region of the L protein appears to be important in cell attachment and consequently in viral infectivity, since preS1 synthetic peptides and corresponding antibodies inhibit virus binding to HepG2 cells (3,4).A cellular receptor is required for HBV binding and penetration in liver cells. Studies in rat hepatoma cells transfected with the HBV genome (5) and transgenic mice with the genome integrated (6) indicate that HBV is able to replicate in rodent cells once it bypasses the attachment and entry steps of infection, so that the absence of specific cellular receptor(s) constitutes the barrier of HBV infection and replication in nonhuman hepatocytes.…”

mentioning

confidence: 99%

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confidence: 99%

“…HBV particles bind to HepG2 cells in a saturable manner but are refractive to virus replication, probably due to the inability of the incoming virus to reach the nucleus in order to gain access to the cellular transcriptional machinery (25). The binding of viral particles to HepG2 may be prevented by HBV preS1-(21-47) peptide or anti-preS1-(21-47) antibodies (3,4,8), suggesting that this sequence could be directly responsible for HBV attachment to the cellular surface (26).…”

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confidence: 99%

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Cloning and Expression of a Novel Hepatitis B Virus-binding Protein from HepG2 Cells

Falco¹,

Ruvoletto²,

Verdoliva³

et al. 2001

Journal of Biological Chemistry

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The hepatitis B virus (HBV), 1 a member of the hepadnaviruses family, is able to infect only humans or higher primates and has a strong organ tropism for hepatocytes (1). The mechanism of hepatocyte infection induced by this noncytolytic virusis not yet clear. The infectious unit of HBV is an enveloped virion of 42-nm diameter that contains an icosahedral nucleocapsid that encompasses a circular, partly double-stranded DNA molecule with a single strand region of variable length, a DNA-linked protein with functions critical for packaging and DNA replication, including priming, RNA-and DNA-dependent DNA polymerase, and RNAH activities. The HBV envelope that determines the targeting to the host cells and the early entry steps is composed of three proteins anchored in a lipid bilayer, occurring in mature virions in both glycosylated and nonglycosylated forms. The three proteins are called small (S), middle (M), and large (L) surface HBV antigens (HBsAg). They are the translational products of three overlapping open reading frames that start from different initiation codons localized at the 5Ј end of the preS1, preS2, and S regions of the env gene (2). The role of the preS1 region of the L protein appears to be important in cell attachment and consequently in viral infectivity, since preS1 synthetic peptides and corresponding antibodies inhibit virus binding to HepG2 cells (3,4).A cellular receptor is required for HBV binding and penetration in liver cells. Studies in rat hepatoma cells transfected with the HBV genome (5) and transgenic mice with the genome integrated (6) indicate that HBV is able to replicate in rodent cells once it bypasses the attachment and entry steps of infection, so that the absence of specific cellular receptor(s) constitutes the barrier of HBV infection and replication in nonhuman hepatocytes. A number of putative human cellular receptors for HBV have been proposed, including the receptor for immunoglobulin A (IgA), which shows a partial sequence similarity between the Fc region of the IgA ␣ chain and the preS1 domain of the virus (7,8), and the receptor for interleukin-6, due to the presence of a recognition site for the preS1 domain on interleukin-6

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Section: Hbv-bp Is Responsible For Hbv Entry In Transfected Cells Andmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Cloning and Expression of a Novel Hepatitis B Virus-binding Protein from HepG2 Cells

Falco¹,

Ruvoletto²,

Verdoliva³

et al. 2001

Journal of Biological Chemistry

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“…The virus envelope proteins can be expected to interact with a cellular receptor and all three envelope proteins have been shown to interact directly or indirectly with a range of cellular proteins (De Meyer et al, 1997). The preS1 region binds to HepG2 cells and liver cell membranes Pontisso et al, 1989 ;Qiao et al, 1994), the preS2 domain binds to HepG2 cells, T lymphocytes and liver cell membranes Franco et al, 1992 ;Machida et al, 1983), and the S domain binds to hepatocytes, fibroblasts, mononuclear cells and Vero cells (Komai & Peeples, 1990 ;Leenders et al, 1990). Many potential cellular receptors have been proposed (Budkowska et al, 1993(Budkowska et al, , 1995Dash et al, 1992 ;Franco et al, 1992 ;Hertogs et al, 1993 ;Machida et al, 1983 ;Mehdi et al, 1994 ;Neurath et al, 1992 ;Petit et al, 1992 ;Pontisso et al, 1992 ;Treichel et al, 1994) but it is still unclear whether the authentic receptor has been identified.…”

Section: Author For Correspondence : Eric Gowans (At Sir Albert Sakzementioning

confidence: 99%

A cellular protein which binds hepatitis B virus but not hepatitis B surface antigen.

Harvey

Macnaughton

Park

et al. 1999

Journal of General Virology

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The envelope of hepatitis B virus (HBV) consists of three related proteins known as the large (L), middle (M) and small (S) hepatitis B surface antigens (HBsAg). L-HBsAg has a 108-119 amino acid extension at the N terminus compared with M-HBsAg and contains the preS1 sequence of the HBV envelope. Previous research has identified this region as the likely virus attachment protein which is thought to interact with the cellular receptor for the virus. However, as the receptor has still not been identified unequivocally, we used the preS1 region of L-HBsAg to screen a human liver cDNA library by the yeast two-hybrid system. Several positive clones were isolated which encoded cellular proteins that interacted with the HBV preS1 protein. The specificity was examined in an independent manner in experiments in which baculovirus-derived glutathione S-transferase (GST)-preS1 was incubated with 35 S-labelled protein expressed by in vitro translation from the positive clones. The intensity of the interactions using this alternative approach mirrored those observed in the yeast two-hybrid system and two proteins (an unidentified protein and a mitochondrial protein) were selected for further study. The specificity of the binding reaction between the preS1 protein and these two proteins was further confirmed in a competition assay ; HBV purified from serum, but not purified HBsAg, was able to compete with preS1 and thus block GST-preS1 binding to the unidentified protein but not to the mitochondrial protein. The unidentified protein was then expressed as a fusion protein with GST and this was able to bind HBV virions in a direct manner.

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The precore sequence of hepatitis B virus is required for nuclear localization of the core protein

et al. 1997

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The cellular localization of the precore/core and core proteins was studied by immunofluorescence following transfection of 143 thymidine kinase-negative (TK ) and Hep-G2 cells with expression constructs containing wild-type (hepatitis B e antigen [HBeAg]-positive) and precore mutant (HBeAg-negative) sequences. Precore/core constructs with the wild-type phenotype result in strong nuclear staining, while, in contrast, constructs expressing core antigen alone have strong cytoplasmic staining. These differences in the pattern of immunofluorescence staining may be caused by expression of the precore/core protein, some of which may be translocated into the nucleus, following removal of the signal peptide. In vitro translation experiments showed that the main protein products obtained in the presence of microsomal membranes were the precore/core protein and a truncated product representing the same protein without its signal peptide. Core protein expression from the precore mutant constructs was very much reduced, indicating that translational re-initiation was not very efficient. The significance of the precore/core protein being present in the nucleus is not clear, but suggests that it may be important in the replicative cycle of the virus. Finally, HBeAg produced by some of the constructs could not be detected because amino acid substitutions affected antibody-binding epitopes.

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Adsorption and Penetration of Hepatitis B Virus in a Nonpermissive Cell Line

Cited by 37 publications

References 0 publications

Cloning and Expression of a Novel Hepatitis B Virus-binding Protein from HepG2 Cells

Cloning and Expression of a Novel Hepatitis B Virus-binding Protein from HepG2 Cells

A cellular protein which binds hepatitis B virus but not hepatitis B surface antigen.

The precore sequence of hepatitis B virus is required for nuclear localization of the core protein

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