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Biomedical subjects

T F Deuel

Publications and source records attributed to T F Deuel.

At least 91 records · Page 5Linked to original sources
PubMed1988

Identification and purification of PDGF/sis-like proteins from nuclei of simian sarcoma virus-transformed fibroblasts.

The platelet-derived growth factor (PDGF) is a potent mitogen for cells derived from mesenchyme and is highly related to the transforming protein of the simian sarcoma virus (SSV), p28v-sis. Both PDGF and p28v-sis appear to initiate DNA synthesis and cell proliferation through an interaction with PDGF cell surface receptors but the identity of the PDGF induced signals which are recognized within the nucleus is unknown. It has been suggested that PDGF is directly transported to the nucleus although conflicting data have been obtained when nuclear fractions have been analysed for PDGF-like proteins. Specific antisera recognizing PDGF and p28v-sis were used to isolate and partially characterize proteins from nuclei of SSV-transformed NRK fibroblasts. Nuclear proteins of 66, 65, and 44 kD were identified in immunoprecipitates. These proteins were displaced competitively from binding to anti-PDGF antisera by either PDGF or by mitogenically active recombinant v-sis protein homodimers and the proteins were not recognized by non-immune sera. Proteins of 66, 65, and 44 kD also were partially purified from nuclear extracts with anti-PDGF immunoaffinity chromatography and were identified in silver stained PAGE gels. The data establish the presence of proteins antigenically related to PDGF within the nuclei of SSV-transformed cells and suggest possible roles of nuclear proteins related antigenically to PDGF and p28v-sis in cell growth and transformation.

Animals↗
PubMed1988

Platelet-derived growth factor: a multifunctional regulator of normal and abnormal cell growth.

Polypeptide growth factors, which are synthesized and secreted both by normal and transformed cells, have a multiplicity of roles in normal and abnormal cell growth. Platelet-derived growth factor (PDGF) is nearly identical to the transforming gene (oncogene) product of simian sarcoma virus p28v-sis. Binding of PDGF to high-affinity cell surface PDGF receptors stimulates the intrinsic autophosphorylating protein tyrosine kinase activity of these receptors and sets off a cascade of metabolic events. Among these events are induction of otherwise quiescent genes which regulate synthesis of transcriptional factors and thus are critical in cell division, differentiation and development.

Animals↗
PubMed1987

Monoclonal antibody C3.1 is a platelet derived growth factor (PDGF) antagonist.

A monoclonal antibody C3.1 raised against NR6 cells and partially purified PDGF receptor blocked PDGF stimulated [3H] thymidine incorporation in Swiss mouse 3T3 cells and immunoprecipitated a 180 kDa phosphoprotein from NR6 cells. The phosphoprotein bound to a C3.1 sepharose 4B affinity matrix and could be specifically eluted with PDGF but not by EGF or basic FGF. These preliminary results suggests that the ability of C3.1 to inhibit PDGF stimulated mitogenesis may be due to its direct or allosteric interaction at the PDGF receptor binding site.

Animals↗
PubMed1987

Accelerated healing of incisional wounds in rats induced by transforming growth factor-beta.

The role of polypeptide growth factors in the processes of inflammation and repair was investigated by analyzing the influence of transforming growth factor-beta (TGF-beta), applied directly to linear incisions made through rat dorsal skin. A dose-dependent, direct stimulatory effect of a single application of TGF-beta on the breaking strength of healing incisional wounds was demonstrated. An increase in maximum wound strength of 220 percent of control was observed at 5 days; the healing rate was accelerated by approximately 3 days for at least 14 days after production of the wound and application of TGF-beta. These increases in wound strength were accompanied by an increased influx of mononuclear cells and fibroblasts and by marked increases in collagen deposition at the site of application of TGF-beta. TGF-beta is thus a potent pharmacologic agent that can accelerate wound healing in rats.

Animals↗
PubMed1987

Platelet-derived growth factor/sis in normal and neoplastic cell growth.

Platelet-derived growth factor (PDGF) is the major growth factor in serum and a potent mitogen for cells mesenchymal origin. It is a highly basic heterodimeric protein with a molecular mass of approximately 30 kDa and binds to a cell surface receptor with high affinity. The amino acid sequence of PDGF revealed sequence homology to the v-sis gene product of simian sarcoma virus (SSV), a transforming retrovirus. Characterization of cells transformed by SSV has revealed PDGF-related proteins in subcellular organelles and in conditioned media consistent with the autocrine stimulation of cell growth through cell surface receptors and perhaps through an internal autocrine mechanism as the growth factor and its receptor are processed. PDGF is also a potent chemotactic agent for inflammatory and other mesenchymal cells and has been implicated in normal tissue repair processes such as wound healing, as well as in aberrant proliferative processes like atherogenesis.

Animals↗
PubMed1987

Ultrastructural localization of a platelet-derived growth factor/v-sis-related protein(s) in cytoplasm and nucleus of simian sarcoma virus-transformed cells.

The subcellular distribution of v-sis-related protein(s) was analyzed in three simian sarcoma virus (SSV)-transformed cell lines with immunofluorescence and protein A-gold labeling techniques using rabbit polyclonal anti-platelet-derived-growth factor (PDGF) antisera. Antigenically reactive proteins were recognized in subcellular organelles related to protein synthesis and processing, including polyribosomes, endoplasmic reticulum, and the Golgi apparatus, as well as on the cytoplasmic surface of plasma membranes. Prominent immunoreactive proteins were also shown in association with nuclear chromatin in intact cells and in isolated nuclei using Lowicryl K4M resin embedding techniques. Protein A-gold labeling was markedly reduced in sections of non-SSV-transformed fibroblasts incubated with anti-PDGF and absent from SSV-transformed cells if Epon resin was substituted for Lowicryl in the embedding process or if sections were with irrelevant antisera. Nuclear localization of v-sis-related antigens was confirmed in a nitrocellulose-based immunoassay using nuclei isolated from SSV-transformed fibroblasts. Thus, polypeptides recognized antigenically as related to the v-sis gene product not only may be found in subcellular organelles associated with protein synthesis and packaging but also may be found in the nucleus of SSV-transformed cells. These results raise the possibility that v-sis- or PDGF-like proteins may function within the nucleus of SSV-transformed cells.

Animals↗
PubMed1987

Polypeptide growth factors: roles in normal and abnormal cell growth.

An increasing number of polypeptide growth factors have been identified that regulate not only cell proliferation but an extraordinary range of cell activities, including matrix protein deposition and resolution, the maintenance of cell viability, cell differentiation, inflammation, and tissue repair. Normal cells appear to require growth factors for proliferation and for maintenance of viability. Cells that secrete a polypeptide growth factor have an advantage in growth. These factors can act either externally through cell surface receptors or perhaps internally during the transport of receptors and growth factors through the ER and Golgi, causing autocrine stimulation of cell growth. Depending on the cell type, growth factors can also be potent inhibitors of cell growth rather than stimulating growth, and the effects can depend on the presence or absence of other growth factors. Platelet-derived growth factor has been shown to be nearly identical to the product of the v-sis gene of the simian sarcoma virus, which appears to cause cell transformation through its interactions with the PDGF receptor activating the tyrosine kinase activity of the PDGF receptor. Similarly, two proto-oncogenes, c-erbB and c-fms, encode growth factor receptors. The EGF receptor activity of the v-erb oncogene product appears to be constitutively activated without the need for growth factor, perhaps because of the truncation at the amino terminus deleting the EGF binding domain. The induction of the myc and the fos proteins by growth factor stimulation of quiescent cells, as well as the potential for the p21 product of the ras oncogene to act as an intermediate in transducing adrenergic signals, provide direct evidence that these pathways are important for stimulation of cell growth. Cells transformed by the v-sis oncogene always appear to bear PDGF cell surface receptors, which suggests that this oncogene has a specific requirement of the PDGF receptor for transformation. In contrast, cells transformed by the v-erbB and v-fms oncogenes are not stimulated by EGF or by CSF-1. Thus it seems likely that the tyrosine kinase activity of the corresponding receptor is ubiquitously expressed in these cases. Major questions remain unanswered. In particular, what are the mechanisms by which growth factors initiate pathways leading to DNA synthesis? What are the physiological substrates of the growth factor receptor tyrosine kinase? Considerable effort also is needed to further define the cellular specificity of the different growth factors, particularly within intact tissues, and to determine how the various growth factors interact.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Diminished response of Werner's syndrome fibroblasts to growth factors PDGF and FGF.

Patients with Werner's syndrome, an autosomal recessive disorder, undergo an accelerated aging process that leads to premature death. Fibroblasts from such patients typically grow poorly in culture. Here it is shown that fibroblasts from a patient with Werner's syndrome have a markedly attenuated mitogenic response to platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF). In contrast, they have a full mitogenic response to fetal bovine serum. Both PDGF binding and receptor numbers per cell are unaltered. The Werner's syndrome cells express high constitutive levels of collagenase in vitro. Although PDGF enhances collagenase expression through increased levels of hybridizable collagenase messenger RNA in normal skin fibroblasts, no induction of collagenase occurs in the Werner's syndrome fibroblasts. Moreover, the failure to respond to this agonist effect of PDGF is not restored by fetal bovine serum. The data suggest that failure of one or more PDGF-mediated pathways in Werner's syndrome cells may contribute to the phenotypic expression of the disorder.

Cell Division↗
PubMed1986

Purification and partial characterization of prostate-derived growth factor.

A potent growth-promoting polypeptide, the prostate-derived growth factor (PrDGF), has been purified to apparent homogeneity from acid extracts of rat prostatic tissue using ion-exchange, reverse-phase, and gel-permeation chromatography. PrDGF migrates as a single protein-staining band in NaDodSO4/PAGE in precise correspondence to extractable PrDGF activity in nonstained NaDodSO4 gels. PrDGF is acid- and heat-stable but is sensitive to reduction or protease treatment. PrDGF is an acidic (pI 5.0) protein of approximately equal to 25 kDa in NaDodSO4/polyacrylamide gels and of approximately equal to 6-8 kDa in reduced NaDodSO4/polyacrylamide gels. PrDGF stimulates the linear incorporation of [methyl-3H]thymidine into normal rat kidney cells between 0 and 16 ng/ml. PrDGF appears to differ from other known growth factors in chemical composition and biological properties, suggesting that PrDGF is a previously undescribed growth factor.

Animals↗
PubMed1986

Isolation and sequencing of a cDNA clone homologous to the v-sis oncogene from human endothelial cells.

A clone containing the 3' end of the mRNA for the human c-sis gene (homologous to the B chain of platelet-derived growth factor) was isolated from a cDNA library derived from human umbilical vein endothelial cells and then sequenced. The analysis of possible translation products in all three reading frames indicated that the A chain of platelet-derived growth factor was not coded for within the 3' end of the c-sis mRNA. The 3' end of the mRNA for c-sis is contained in or adjacent to exon 6.

Base Sequence↗
PubMed1985

Stimulation of in vitro human skin collagenase expression by platelet-derived growth factor.

Platelet-derived growth factor (PDGF) is both chemoattractant and mitogenic for stromal cells. Here, we examined the effects of PDGF on collagenase expression by normal human skin fibroblasts. Culturing cells for 24 hr in the presence of PDGF at 0-180 ng/ml resulted in a dose-dependent, saturable increase in collagenase activity in the culture medium that was paralleled by equal increases in immunoreactive collagenase protein, suggesting enhanced synthesis of a catalytically unaltered enzyme. The specificity of this effect was demonstrated by comparing the collagenase-stimulatory effect with that on total protein synthesis and DNA synthesis. Under in vitro conditions that produced a 2.5-fold increase in collagenase synthesis, there was an approximately equal to 20% increase in total protein synthesis and no change in DNA synthesis. In addition, platelet factor 4, another platelet-derived protein, caused a less than 20% increase in collagenase expression. In time-course studies, stimulation of collagenase synthesis was first observed 8-10 hr after exposure to the growth factor. Conversely, when cells were primed with PDGF for approximately equal to 24 hr and the stimulator was then removed, an increased rate of synthesis was seen for an additional approximately equal to 6 hr, after which the rate reverted to control levels. Since the kinetic data suggested a possible pretranslational effect, fibroblasts cultured with PDGF were used to prepare mRNA. In cell-free translation, total protein synthesis was essentially unaltered; however, the growth factor caused a greater than 2-fold increase in translatable collagenase mRNA. The data suggest that PDGF specifically modulates collagenase synthesis, possibly through a series of events that lead to increased transcription or preferential translation of collagenase mRNA.

Cells, Cultured↗
PubMed1985

Dissociation of the chemotactic and mitogenic activities of platelet-derived growth factor by human neutrophil elastase.

Because platelet-derived growth factor (PDGF) may be released at sites where neutrophil proteinases may also be released, we examined the effects of neutrophil elastase and cathepsin G upon the chemotactic and mitogenic activities of PDGF. Elastase abolished the chemotactic activity of PDGF for fibroblasts but had no effect on its chemotactic activity for monocytes, or on its mitogenic activity for 3T3 cells or its capacity to bind to 3T3 cells. Cathepsin G had no effect upon the chemotactic or mitogenic activities of PDGF. In contrast, trypsin eliminated the chemotactic activity of PDGF for monocytes and fibroblasts and the mitogenic activity of PDGF. After reduction and alkylation, PDGF retained full chemotactic activity for fibroblasts and monocytes but exhibited no mitogenic activity and only limited binding to 3T3 cells. These results indicate separate domains on PDGF for fibroblast chemotactic and mitogenic activity and for monocyte and fibroblast chemotactic activity and raise the possibility that the biological activities of PDGF may be modified selectively in vivo. The findings further suggest that the majority of PDGF receptors on fibroblasts mediate mitogenic activity and that only a minority of the PDGF receptors on fibroblasts are responsible for chemotactic activity.

Alkylation↗
PubMed1985

Platelet-derived growth factor: roles in normal and v-sis transformed cells.

Platelet-derived growth factor (PDGF) is the most potent mitogenic protein in human serum. The normal roles of this protein may relate to its potent mitogenic properties and its activities in directed cell migration and at sites of wounds. PDGF is a heterodimeric protein of approximately 30,000 molecular weight; one polypeptide chain of PDGF is highly homologous to the predicted amino acid sequence of p28v-sis, the putative transforming protein of simian sarcoma virus (SSV), suggesting a major role of growth factor activity in transformation by SSV. PDGF-like growth promoting activity is found in SSV-transformed cells and is secreted into conditioned media where it appears to interact with PDGF cell surface receptors to stimulate 3H-thymidine incorporation into DNA of cells secreting this protein. Transformation by SSV may in part be mediated by the autocrine stimulation of cell growth by the PDGF-like mitogenic properties of the transforming protein of SSV.

Animals↗
PubMed1985

Platelet-derived growth factor promotes human peripheral monocyte activation.

Like in the polymorphonuclear leukocyte (PMN), the platelet-derived growth factor (PDGF) purified to homogeneity is capable of inducing monocyte activation responses as evaluated by generation of superoxide anion (O-.2) from membrane-associated oxidase system, release of granule enzymes, and enhanced cell adherence and cell aggregation. Superoxide anion release was maximized at 10 ng/mL PDGF and was comparable to that induced by 10(-7) mol/L formyl-methionyl-leucyl-phenylalanine. The potency of PDGF to induce this response in monocytes was of the same magnitude as that observed in PMNs. Similarly, lysozyme release and monocyte adherence were also increased in a dose-dependent manner and achieved maximal responses at 40 ng/mL concentration of PDGF. The PDGF concentration required to achieve maximal monocyte aggregation was two-fold (60 ng/mL) of that found for PMNs. In contrast to PMNs, a positive correlation (gamma = .93; P less than .01) was observed between the increases of PDGF concentration and beta-glucuronidase release. These findings indicate that PDGF can induce the full sequence of cell activation events in human monocytes similar to human PMNs.

Cell Adhesion↗
PubMed1984

The c-sis gene encodes a precursor of the B chain of platelet-derived growth factor.

The relationship between platelet-derived growth factor (PDGF) and the proto-oncogene c-sis has been determined by amino acid sequence analysis of PDGF and nucleotide sequence analysis of c-sis genomic clones. The nucleotide sequences of five regions of the human c-sis gene which are homologous to sequences of the transforming region (v-sis) of simian sarcoma virus (SSV) were determined. By alignment of the c-sis and v-sis nucleotide sequences the predicted amino acid sequence of a polypeptide homologous to the putative transforming protein p28sis of SSV was deduced. Both predicted sequences use the same termination codon and additional coding sequences may lie 5' to the homologous regions. Amino acid sequence analysis of the PDGF B chain shows identity to the amino acid sequence predicted from the c-sis sequences over 109 amino acid residues. Polymorphism may exist at two amino acid residues. These results suggest that c-sis encodes a polypeptide precursor of the B chain. A partial amino acid sequence of the PDGF A chain is also described. This chain is 60% homologous to the B chain and cannot be encoded by that part of c-sis which has been sequenced but could be encoded by sequences which lie 5' to the five regions of v-sis homology in c-sis, or at a separate locus.

Amino Acid Sequence↗