A small v-sis/platelet-derived growth factor (PDGF) B-protein domain in which subtle conformational changes abrogate PDGF receptor interaction and transforming activity.

Giese, N; LaRochelle, William J.; May-Siroff, M; Robbins, K C; Aaronson, Stuart A.

doi:10.1128/mcb.10.10.5496

Cited by 19 publications

(12 citation statements)

References 29 publications

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“…The identification of specific residues in PDGF that are responsible for receptor activation has been intensively investigated by several laboratories (29)(30)(31)(32)(33). The (35).…”

mentioning

confidence: 99%

Cellular transformation by a transmembrane peptide: structural requirements for the bovine papillomavirus E5 oncoprotein.

Meyer

Webster

et al. 1994

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

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The ES oncoprotein of bovine papillomavirus, only 44 amino adds long, occurs as a disulfide-bonded transmembrane dimer. This remarkable oncoprotein stimulates signal transduction.through activation of the plateletderived growth factor (PDGF) receptor, and E5 exhibits limited amino acid sequence similarit with PDGF. Results presented here sugest that a key feature of the hydrophobic transmembrane domain is an amino acid side chain that participates in interhelical hydrogen bond formation. These data are reminiscent of the activated neu oncogene, in which a point mutation in the membrane domain leads to lipndindependent dimerization and activation of a receptor tyrosine kinase. Signfcantly, the tranmembrane domain of E5 can be largely replaced by the transmembrane domain from the activated neu receptor tyrosine kinase. Extensive mutageneis defines the minimal structural features required for transformation by the ES oncoprotein as, first, the ability to dimerize and, second, presentation ofa negatively charged residue at the extralular side of the membrane. The biological activity of ES mutants that lack most amino acid residues similar to PDGF snggests that ES and PDGF activate the PDGF receptor by disnct mechanisms.Bovine papillomavirus type I (BPV) typifies the fibropapilloma viruses, which transform fibroblasts in vitro and lead to development of fibrosarcomas in their host. In contrast, the human papillomaviruses infect epithelial cells and are strongly implicated in carcinomas of the cervix and respiratory tract (1). The fibroblast-transforming function of BPV is due to a small open reading frame, designated E5, encoding a 44-residue protein that forms membrane-associated disulfide-bonded dimers (2-6). Recent work has demonstrated that the E5 oncoprotein activates the platelet-derived growth factor (PDGF) P-receptor (7,8) and may display functional similarity with the human T-lymphotropic virus type I p12' protein (9).Inspection of the amino acid sequence of the E5 oncoprotein reveals two distinct domains (2-6): an extremely hydrophobic N-terminal domain, residues 1-32, and a hydrophilic C-terminal domain, residues 33-44. Several studies have led to the conclusion that the E5 oncoprotein is membraneanchored with a type II orientation, with the N terminus intracellular and the C terminus extracellular (10). In addition, the hydrophobic domain of E5 can function as a signal-anchor domain, indistinguishable from signal-anchor domains of well-characterized type II proteins such as neuraminidase, transferrin receptor, or asialoglycoprotein receptor (11).In this work, we describe the minimal structural features required for biological transforming activity by a transmembrane peptide. The results presented here suggest that E5 and PDGF activate the PDGF receptor by fundamentally distinct mechanisms. ES/neu Substitution Mutant. Degenerate oligonucleotides were synthesized that would encode a pool of E5/neu transmembrane recombinants in which the 44 residues would be derived as follows: 1-5 from E5, 6-10 from...

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“…The identification of specific residues in PDGF that are responsible for receptor activation has been intensively investigated by several laboratories (29)(30)(31)(32)(33). The (35).…”

mentioning

confidence: 99%

Cellular transformation by a transmembrane peptide: structural requirements for the bovine papillomavirus E5 oncoprotein.

Meyer

Webster

et al. 1994

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

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“…As shown in Fig. 8, this window of minimal phylogenetic drift corresponds roughly to "B-chain active sites" which have been mapped by mutational analysis Giese et al, 1990;Ostman et al, 1991). The B-chain active site defines the universal PDGF agonist.…”

Section: Discussionmentioning

confidence: 95%

“…Operationally, the B-chain active site is a sequence of amino acids within PDGF B which, in chimeric molecules, confers upon PDGF A subunits the ability to interact with either alpha or beta receptor subunits. It is of interest that mutational analysis Giese et al, 1990;Ostman et al, 1991) and phylogenetic analysis (Fig. 8) converge independently upon the same set of amino acids as being especially critical to receptor activation.…”

Section: Discussionmentioning

confidence: 98%

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Recombinant PDGF from Lower Vertebrates: Receptor Binding and Immunochemical Analysis with Metabolically Labeled Growth Factor

1992

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We used a baculovirus vector/insect host cell system to express cDNA clones of PDGF A genes from mouse and frog (Xenopus laevis). The insect host cells process PDGF A subunits from either frogs or mice into biologically active AA homodimers with yields in the range of 0.5-1.0 mg/liter of culture medium. The recombinant PDGFs can be metabolically labeled with 35S-cysteine for use in radioreceptor and radioimmunoassays. Neutralizing polyclonal antisera can be raised against the mouse and frog PDGFs. These antisera are markedly species-specific in action. However, in radioreceptor binding assays and bioassays for mitogenic activity, human, mouse and frog PDGF AA homodimers occupy and activate murine PDGF receptors with equal efficiency.

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“…Molecular genetic analysis has already identified at least two and possibly three independent sites in the PDGF B molecule that specify highaffinity interaction with the p PDGFR (LaRochelle et al, 1990Giese et al, 1990;Ostman et al, 1991b;Maher et al, 1993). PDGF A and B sites of interaction with the a PDGFR have yet to be elucidated.…”

Section: Discussionmentioning

confidence: 96%

Platelet-derived growth factor AB heterodimer interchain interactions influence secretion as well as receptor binding and activation

May

Aaronson²,

LaRochelle³

1993

Biochemistry

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Platelet-derived growth factor (PDGF) is a disulfide-linked dimer comprised of two related polypeptide chains. To investigate the effects of an inactivating lesion introduced into one chain of the nascent PDGF dimer, approaches were developed to optimize synthesis, assembly, secretion, and purification of heterodimers between normal PDGF A and wild-type or mutant PDGF B. PDGF AB heterodimers were released into culture fluids less efficiently than PDGF AA, but to a greater degree than the cell-associated PDGF BB. These results suggest that interactions between two chains influence PDGF secretion. Analysis of heterodimers between PDGF A and disabled PDGF B mutants on cells that express either alpha or beta PDGFRs demonstrated that the impaired biologic activity of the mutant PDGF B chain was ameliorated with respect to binding and triggering of alpha PDGFRs. In cells that expressed both receptor types, heterodimers of mutant PDGF B and wild-type PDGF A gained substantially in their ability to recruit and trigger alpha, but not beta, PDGFRs. Partial rescue of impaired PDGF B mutant chain function by dimer formation with a wild-type PDGF A chain implies that interchain interactions markedly affect PDGFR binding and activation.

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A small v-sis/platelet-derived growth factor (PDGF) B-protein domain in which subtle conformational changes abrogate PDGF receptor interaction and transforming activity.

Cited by 19 publications

References 29 publications

Cellular transformation by a transmembrane peptide: structural requirements for the bovine papillomavirus E5 oncoprotein.

Cellular transformation by a transmembrane peptide: structural requirements for the bovine papillomavirus E5 oncoprotein.

Recombinant PDGF from Lower Vertebrates: Receptor Binding and Immunochemical Analysis with Metabolically Labeled Growth Factor

Platelet-derived growth factor AB heterodimer interchain interactions influence secretion as well as receptor binding and activation

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