Immunological characterization of a major transformation-sensitive fibroblast cell surface glycoprotein. Localization, redistribution, and role in cell shape.

Yamada, Kenneth M.

doi:10.1083/jcb.78.2.520

Cited by 171 publications

(65 citation statements)

References 55 publications

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“…FN netoworks were detected on the surface of the substratum that surrounds the point of attachment of the mitotic cells by the immunofluorescence method. Similar observations have been reported for some populations of mitotic cells (19) and for virus-transformed cells (21,24). These observations taken together indicate that mitotic cells become round leaving FN in networks on the substratum.…”

supporting

confidence: 75%

Stripping of Extenral Cell-Associated Fibronectin from Mitotic Fibroblasts

Iwashita¹,

Yahara²

1981

Cell Struct. Funct.

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ABSTRACT. Using a sensitive method to determine the amount of fibronectin (FN), we investigated the release of cellular FN in C3H mouse fibroblasts, which depends upon the progress of the cell cycle. Cells first labeled with [14C]leucine and arrested in confluent culture were trypsinized and replated in fresh medium containing [14C]leucine with or without 1 mM hydroxyurea (HU). The cell number in the untreated culture increased 1.3-fold 36 h after replating, whereas HU-treated cells neither entered the S phase nor divided. The synthesis, turnover and rebinding of FN to cells were almost the same for both cultures. The amounts of FN released into the medium also were the same for untreated and HU-treated cultures. These results indicate that FN was not released into the medium specifically before or during the rounding up of mitotic cells. Immunofluorescence observations using a specific antibody directed against FN showed the presence of FN networks on the surface of the substratum, which surrounded the point at which mitotic cells were attached. In addition, for about half the population of mitotic cells no FN was detected on the cell surfaces. Because of these results, we suggest that when mitotic cells become round, they leave FN in networks on the substratum.Fibronectin (FN) is a high molecular weight glycoprotein present on the surfaces of cells, in connective tissues, and in extracellular fluids (20,23). One function of this protein is to mediate cell adhesion (25). An addition of FN to transformed cells increases their adhesiveness to the substratum, an adhesiveness that had decreased because of transformation; it therefore restores normal cell morphology (2, 25). Treatment of cultured fibroblasts with trypsin induces the rounding of these cells, probably because FN and other adhesive proteins have been lost due to trypsin treatment (12). It also has been reported that cytochalasin B induces the release of FN into medium, which may be related to alterations in cell morphology after treat-* Present address:

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supporting

confidence: 75%

Stripping of Extenral Cell-Associated Fibronectin from Mitotic Fibroblasts

Iwashita¹,

Yahara²

1981

Cell Struct. Funct.

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“…The diminished FN matrix in most transformed cells may be because of deficient FN synthesis, cell surface binding, assembly, or FN proteolysis (Wagner et al, 1981;Yamada, 1978;Fegan et al, 1981;Chen and Chen, 1987) but probably not to defective FN (Wagner et al, 1981). In SV40 transformed WI-38 fibroblasts, deficient amino terminal binding appears to be important (McDonald et al, unpublished data).…”

Section: Disorganization Of the Fnr In Sv40 Transformed Fibroblasts Imentioning

confidence: 99%

The fibronectin receptor is organized by extracellular matrix fibronectin: implications for oncogenic transformation and for cell recognition of fibronectin matrices.

Roman

Lachance

Broekelmann

et al. 1989

The Journal of Cell Biology

145

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Abstract. Cells interact with extracellular fibronectin (FN) via adhesive fibronectin receptors (FNRs) that are members of the very late antigens (VLAs) subgroup of the integrin family. In stationary fibroblasts, the FNR is highly organized and distributed identically to extracellular FN fibrils. However, in highly migratory neural crest cells and embryonic somatic fibroblasts, this organization is lost and the FNR appears diffuse. Similarly, oncogenic transformation typically leads to disorganization of the FN receptor and loss of matrix FN. Two models can account for these observations. First, the FN matrix may organize the FN receptor at extracellular matrix contacts on the cell surface. Motile cells not depositing FN matrices thus lack organized receptors. Alternatively, as the FNR is required for optimal FN matrix assembly, (McDonald, J. A., B. J. Quade, T. J. Broekelmann, R. LaChance, K. Forseman, K. Hasegawa, and S. Akiyama. 1987. J. Biol. Chem. 272:2957-2967; Roman, J., R. M. LaChance, T. J. Broekelmann, C. J. R. Kennedy, E. A. Wayner, W. G. Carter, J. A. McDonald. 1989. J. Cell Biol. 108:2529-2543 and has putative cytoskeletal links, it could be organized from within the cell helping to position newly forming FN fibrils. To study this question, we developed peptide antibodies specifically recognizing the a5 subunit of the FNR.Using these antibodies, we examined the organization of FN and of the FNR in normal, matrix assembly inhibited, and SV40-transformed human fibroblasts. On FN-coated substrates, the FNR is found in focal contacts rather than diffusely on the basal cell surface, suggesting FNR interaction with intracellular components. However, when FN fibrils are deposited, the FNR is co-distributed with these fibrils. Preventing FN matrix assembly prevents organization of the FNR. Moreover, when fibroblasts with well established FN matrices and co-distributed FNR are incubated briefly with monoclonal antibodies that block FNR binding to FN, the FNR is no longer co-distributed with the FN matrix. Thus, the FN receptor is organized in fibrils on the cell surface in response to extracellular FN. Because exogenous FN restores a FN matrix and receptor organization to SV40-transformed cells, the diffuse FN receptor phenotype appears to be related to loss of the FN matrix rather than to impaired FNR function. These results explain diffusely distributed FNRs in migratory neural crest and embryonic fibroblasts lacking well organized FN matrices and emphasize the existence of separate but related systems controlling FN deposition and recognition by receptor-armed cells.

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“…To Immunoprecipitation. Affinity purified anti-fibronectin IgG was prepared as previously described (12). Double antibody immunoprecipitation of S-methionine-labeled fibronectin from in vitro translations was performed exactly as described by Adams et al (4,5).…”

Section: Introductionmentioning

confidence: 99%

Partial purification and characterization of the messenger RNA for cell fibronectin

Fagan¹,

Yamada²,

Crombrugghe³

et al. 1979

Nucl Acids Res

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Fibronectin mRNA has been partially purified by guanidine extraction, oligo-(dT)-cellulose chromatography and sucrose density gradient centrifugation. We obtain a fraction which programs a wheat germ in vitro translation system to synthesize a polypeptide species which co-electrophoreses with fibronectin in SDS-polyacrylamide gels and which is immunoprecipitated with affinity purified fibronectin-specific IgG. Analysis of this RNA fraction by methyl mercury hydroxide-agarose gel electrophoresis reveals the presence of a band accounting for 30 percent to 50 percent of the ethidium bromide-staining material in the fraction. The RNA of this band has an estimated molecular weight of about 3 million daltons and is greatly reduced in the corresponding RNA fraction from RSV transformed CEF. This RNA has been tentatively identified as fibronectin mRNA.

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Immunological characterization of a major transformation-sensitive fibroblast cell surface glycoprotein. Localization, redistribution, and role in cell shape.

Cited by 171 publications

References 55 publications

Stripping of Extenral Cell-Associated Fibronectin from Mitotic Fibroblasts

Stripping of Extenral Cell-Associated Fibronectin from Mitotic Fibroblasts

The fibronectin receptor is organized by extracellular matrix fibronectin: implications for oncogenic transformation and for cell recognition of fibronectin matrices.

Partial purification and characterization of the messenger RNA for cell fibronectin

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