Antibodies to Synthetic Peptides Targeted to the Transforming Genes of Human Adenoviruses: An Approach to Understanding Early Viral Gene Function

Green, Maurice; Brackmann, Karl H.; Lucher, Lynne A.; Symington, Janey

doi:10.1007/978-3-642-69460-8_8

Cited by 6 publications

(3 citation statements)

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“…The E1B 19 kDa protein is expressed within the first 10 h post-infection (Green et al, 1983;Wold and Gooding, 1991;Akusjä rvi, 1993). We therefore speculate that it might be especially important for the virus to counteract NF-B activity early in the course of the viral infection in order to escape an antiviral response.…”

Section: Discussionmentioning

confidence: 99%

A Hydrophobic Region within the Adenovirus E1B 19 kDa Protein Is Necessary for the Transient Inhibition of NF-κB Activated by Different Stimuli

Limbourg

Stadtler

Chinnadurai

et al. 1996

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 99%

A Hydrophobic Region within the Adenovirus E1B 19 kDa Protein Is Necessary for the Transient Inhibition of NF-κB Activated by Different Stimuli

Limbourg

Stadtler

Chinnadurai

et al. 1996

Journal of Biological Chemistry

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“…human 81-globin and e-globin genes (Allan et al, 1984). However, analysis of the effect of adenovirus E1A on several host cell genes, including the genes for pre-proinsulin and globins, suggests that stimulatory activity is restricted to newlyintroduced genes, and does not occur with endogenous class II genes (Babich et al, 1983;Gaynor et al, 1984;Green et al, 1983). This also applies to genes transcribed by polymerase III.…”

Section: Experimental Approachesmentioning

confidence: 99%

Expression and interactions of human adenovirus oncoproteins

Boulanger

Blair

1991

Biochemical Journal

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INTRODUCTION: CELL TRANSFORMATION BY ONCOGENIC DNA VIRUSES Cell growth is a complex process resulting from a cascade of events regulated by both positively and negatively acting biological signals. Tumorigenic proliferation has been shown in some cases to be caused by alterations in genes encoding proteins involved in such signalling pathways (Klein, 1988). DNA tumour viruses have been extensively studied and widely used as molecular probes and models of cellular transformation. The oncogenes, i.e. tumour-inducing genes, from polyomavirus (Py), simian virus 40 (SV40) and adenovirus (Ad), are capable of inducing several distinct changes in cellular phenotype, including immortalization, secretion of growth factors, loss of contact inhibition, anchorage-independent growth and morphological transformation (Branton et al., 1985). However, unlike the transforming retroviruses, the DNA tumour viruses contain oncogenes with apparently no cellular counterparts (Bishop, 1985; Branton et al., 1985; Klein, 1988). Moreover, the proteins encoded by DNA viral oncogenes play an essential role in the lytic life-cycle of the virus in fully permissive host cells (Alwine et al., 1977; Harrison et al., 1977; Lassam et al., 1978), whereas transforming oncogenes of retroviruses are not required for lytic growth (Bishop, 1982). Adenovirus, like the other DNA tumour viruses, encodes a set of proteins that disturb cellular regulatory functions, thus reprogramming cell growth (Berk, 1986a; Branton et al., 1985; Branton & Rowe, 1985). The proteins encoded by oncogenes are termed 'oncoproteins'. Certain viral oncoproteins have been found to form stable complexes with host-cell proteins which also appear to be involved in transformation. This is the case for the Py middle T-pp60-8rc complex (Courtneidge & Smith, 1983), the SV40 large T-p53 complex (Lane & Crawford, 1983; Linzer & Levine, 1979; McCormick & Harlow, 1980), and the adenovirus E1B-p55:p53 complex (Sarnow et al., 1982a). The formation of these complexes may result in the potentiation of the transforming functions of these cellular proteins. The molecular mechanism might involve the alteration of a catalytic activity of the cellular protein, such as the protein kinase activity of pp6Oc-8rc (Bolen et al., 1984; Courtneidge, 1985), or a change in protein conformation and stability (e.g. p53; Oren et al., 1981; Reich et al., 1983). Cellular transformation by the adenovirus oncogenes involves a very elaborate mechanism at the molecular level. It seems to occur as a sequential process, involving two blocks of genes, ElA and E1B. EIA alone has the property of immortalizing primary

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“…Procedures for solid-phase peptide synthesis, peptide coupling to keyhole limpet hemocyanin, and rabbit (New Zealand White) immunization were essentially as described previously (20). The anti-peptide antiserum (3 ,ul) and in some cases anti-BLV antiserum (5 ,ul) (4).…”

Section: Methodsmentioning

confidence: 99%

Identification and some biochemical properties of the major XBL gene product of bovine leukemia virus.

Sagata¹,

Tsuzuku-Kawamura²,

Nagayoshi-Aida³

et al. 1985

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

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Using a rabbit antiserum directed against a synthetic oligopeptide whose sequence was deduced from the nucleotide sequence of the XBL gene of bovine leukemia virus, we detected a 38-kDa protein in virus-producing cell lines. In vitro translation of hybrid-selected RNA unequivocally demonstrates that this protein, designated p38(XBL), is indeed encoded by the XBL gene. Unlike the other virus-encoded proteins, however, p38(XBL) resides within the cells without being incorporated into virions. It undergoes no gross posttranslational modifications and has a relatively short half-life (5-6 hr) in vivo. Furthermore, cell fractionation combined with pulse-chase experiment reveals that a significant fraction (more than half) of the p38(XBL) localizes to the nucleus of the infected cell after synthesis. We conclude that the XBL gene of bovine leukemia virus is a functional gene encoding a nonvirion protein p38(XBL), which possibly functions within the nucleus of the infected cell to regulate viral or cellular gene expression. P38(XBL) is presumably translated from a doubly spliced, bicistronic mRNA that has the capability to encode another small polypeptide in a different reading frame.Bovine leukemia virus (BLV), an exogenous replicationcompetent retrovirus in cattle, is a causative agent of enzootic bovine leukosis (1) and is evolutionarily related to human T-cell leukemia viruses types I and II (HTLV-I and HTLV-II) (2-5). BLV has no preferred chromosomal site for integration (6,7) and often lacks the 5' half (but not the 3' half) ofits genome in the leukemic cells (8). Furthermore, although BLV carries no typical oncogene of cellular origin (5, 9), it is capable of transforming cells in vitro (10, 11). It is thus possible that the BLV genome itself, possibly its 3' half, harbors a gene(s) that is responsible for cellular transformation, viral replication, or both (4).We and others have recently determined the nucleotide sequence of the integrated BLV provirus (3,5,12), finding that the BLV genome contains an unidentified region (designated XBL gene) between the env gene and the 3' long terminal repeat (LTR) (5). The major long open reading frame (XBL-I) found in the XBL gene has a typical protein-encoding capability (4), and we predicted that its putative product would be produced by a spliced mRNA (4). X or LOR genes of HTLVs are genes equivalent to the XBL gene of BLV (4) and have recently been shown to encode proteins of 38-42 kDa (13-16).Here, we identify the XBL-I product in BLV-infected cell lines, using a rabbit antiserum raised against a synthetic oligopeptide derived from the XBL-I open reading frame. Furthermore, we describe several biochemical properties of the XBL-I product, including its metabolic stability and subcellular localization, to gain insight into its biological function. MATERIALS AND METHODSCells and Metabolic Labeling. BLV-infected fetal lamb kidney cells (FLK) (17) and bat lung cells (Bat2clI) (18) were used as sources of BLV-producing cells. Cells spread at 60-70% confluency were me...

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Antibodies to Synthetic Peptides Targeted to the Transforming Genes of Human Adenoviruses: An Approach to Understanding Early Viral Gene Function

Cited by 6 publications

References 103 publications

A Hydrophobic Region within the Adenovirus E1B 19 kDa Protein Is Necessary for the Transient Inhibition of NF-κB Activated by Different Stimuli

A Hydrophobic Region within the Adenovirus E1B 19 kDa Protein Is Necessary for the Transient Inhibition of NF-κB Activated by Different Stimuli

Expression and interactions of human adenovirus oncoproteins

Identification and some biochemical properties of the major XBL gene product of bovine leukemia virus.

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