Human fibronectin: molecular cloning evidence for two mRNA species differing by an internal segment coding for a structural domain.

Kornblihtt, Alberto R.; Vibe-Pedersen, Karen; Baralle, Francisco E.

doi:10.1002/j.1460-2075.1984.tb01787.x

Cited by 219 publications

(116 citation statements)

References 28 publications

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“…Previous studies (8,9) have suggested that the ED segment seems to be absent in the hepatocyte mRNAs, which are the source of plasma FN, and it is probable that the inclusion of Here we show that the expression of the IIICS sequence introduces into the FN molecule a region that is susceptible to thermolysin cleavage. This agrees with previous indications of a different susceptibility to proteolytic enzymes in the COOH-terminal region of the two FN subunits (2,11,15,20,23,24,26).…”

Section: Discussionsupporting

confidence: 48%

Transformed human cells release different fibronectin variants than do normal cells.

Castellani

Siri²,

Rosellini³

et al. 1986

The Journal of Cell Biology

120

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Abstract. Fibronectin molecules are dimers composed of subunits whose primary structures may differ. This is due to alternative splicing in at least two regions (ED and IIICS) of the pre-mRNA.Using two monoclonal antibodies specific for two different epitopes of domain 5 (high affinity for heparin), we have quantitatively analyzed the expression of the IIICS sequence in human fibronectins from different sources. The results demonstrated that the percentage of fibronectin subunits containing the IIICS is higher in fibronectins from tumor-derived or simian virus 40-transformed human cells than in fibronectins from human plasma or normal human fibroblasts.Furthermore, we observed that 45-65 % of fibronectin subunits from transformed cells or normal embryonic fibroblasts are sialylated on the heparinbinding domain 5, whereas this occurs in only 24-28% of fibronectin subunits from normal adult fibroblasts. On the contrary, no sialylation was observed on domain 5 in fibronectin from human plasma.F IBRONECTINS (FNs) ~ are high molecular weight adhesive glycoproteins present in soluble form in plasma and other body fluids and in insoluble form in the extracellular matrices and basement membranes. FN molecules act as bridges between the cell surface and extracellular material. In fact, the FN molecules contain a cell-binding site and binding sites for collagen, heparin, ganglioside, and fibrin. Due to their multiple interactions FNs play an important role in diverse biological phenomena including cell adhesion, cell migration, hemostasis and thrombosis, wound healing, and the ability to induce a more normal phenotype in transformed cells (for reviews on distribution, structure, and biological functions see references 1, 3, 4, 13, and 16).Recently it has been demonstrated that the FN polymorphism may be due to alternative splicing schemes since many different mRNAs may originate from the primary transcript of a single gene (5, 7-10, 17, 18, 22) localized on chromosome two (6, 27).Here, using the domain-specific monoclonal antibodies IST-7 and IST-2 we have studied the presence of the IIICS sequence in FN from plasma and from the conditioned media of normal and tumor-derived human cells. Materials and Methods Cell Lines and Monoclonal AntibodiesCultured normal human fibroblast cell lines (LZ, from adult human skin; GM-3651-C, from adult human skin; GM-5386, from embryonic human 1. Abbreviations used in this paper: FN, fibronectin; RIA, radioimmunoassay; SV-40, simian virus 40.skin; WI-38, from embryonic human lung) and transformed cell lines (HT-1080, from a human fbrosarcoma; RD, from an embryonic human rhabdomyosarcoma; IgR3, from a human melanoma; and WI-38VA13, simian virus 40 lSV-40]-transformed WI-38 cells) were grown in Eagle's minimum essential medium supplemented with 10% fetal calf serum (Flow Laboratories, Irvine, Scotland) that had been depleted of bovine FN by passage through a large capacity gelatin-Sepharose column.Monoclonal antibodies to human plasma FN were prepared as previously described (25). Partial ...

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Section: Discussionsupporting

confidence: 48%

Transformed human cells release different fibronectin variants than do normal cells.

Castellani

Siri²,

Rosellini³

et al. 1986

The Journal of Cell Biology

120

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“…Recent studies using monoclonal antibodies have emphasized the heterogeneity of laminin in different sites in embryonic and adult tissues (Wan et al, 1984). This raises the possibility that the laminin molecule synthesized in the brain is different from the basement membrane laminin and that variants or isoforms of laminin, similar to those described for fibronectin (Tamkun et al, 1984;Kornblihtt et al, 1984), exist and exert the different functions. In this context, I note a previous report by Bignami et al (1984) saying the laminin-positive structures in rat embryonic brain are confined to the external basal laminae and the blood vessels.…”

Section: Resultsmentioning

confidence: 99%

Do neurons in the vertebrate CNS migrate on laminin?

Liesi¹

1985

The EMBO Journal

240

135

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In adult rat brain the extracellular matrix glycoprotein, laminiin, is found only in basement membranes, but is transiently expressed by astrocytes after brain injury. Here, I show that lanminin also appears in immature brain cells during CNS development, and that its presence coincides with phases of neuronal migration. In early embryos, laninin is seen throughout the whole thickness of the forniing brain, and is apparently synthesized by the cells, as judged by its intracytoplasmic localization. As development proceeds, intracellular laminin becomes restricted to the periventricular regions while punctate deposits of laminin follow the course of vimentin-positive radial glial fibers. In most brain regions, the adult pattern of laminin expression is achieved by birth. In the post-natal rat cerebellum, however, lamniin is detected in external granule cells, in Purkinje cells, and in punctate deposits along the radial Bergmann glial fibers. By day 24 after birth, when the migration of external granule cells is complete, all laminin immunoreactivity disappears from these structures. The transient expression of laniinin in regions where neurons are migrating raises the possibility that lanin plays a role in neuronal migration during CNS development.

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“…Double-stranded cDNA containing the 177-basepair Pst IIPst I fragment coding for the EDA portion of FN was excised from cDNA clone p F H l l l (12). This was ligated into pGEM4Z plasmid (Promega, Madison, WI).…”

Section: Methodsmentioning

confidence: 99%

Eda‐containing fibronectin is synthesized from rheumatoid synovial fibroblast‐like cells

Hino

Shiozawa

Kuroki

et al. 1995

Arthritis & Rheumatism

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Objective. To identify the cells that synthesize EDA-containing fibronectin (FN) and examine the role of EDA+ FN in the pathogenesis of rheumatoid joint lesions.Methods. Localization of EDA+ FN and c-Fos protein in rheumatoid joints was studied immunohistochemically by utilizing antibodies for EDA+ FN and c-Fos. Expression of EDA+ FN was studied by immunoelectron microscopy and in situ hybridization. The amount of EDA+ FN was measured by enzymelinked immunosorbent assay.Results. EDA+ FN was specifically localized in the synovial lining layer of synovium with active rheumatoid arthritis (RA) (n = 17), but not in that with osteoarthritis (n = 4) or with inactive fibrous RA (n = 2).EDA+ FN messenger RNA was localized in the synovial lining layer. EDA+ FN was immunoelectron microscopically localized in the synovial lining fibroblast-like (type B) cells. EDA+ FN was also detected at the cartilagepannus junction and on the surface of RA cartilage. Double staining showed that EDA+ FN colocalized with c-Fos protein in the rheumatoid synovial lining layer. Quantification of EDA+ FN showed that it was highly concentrated in rheumatoid synovial fluids.Conclusion. EDA+ FN is synthesized by the synovial lining fibroblast-like (type B) cells in situ in rheumatoid synovium, and appears to be expressed in association with activated or transformed states of synovium.Fibronectin (FN), synthesized in rheumatoid synovium (1) and specifically located in rheumatoid pannus and on cartilage surface (2,3), may be important in rheumatoid joint destruction (4). FN is composed of more than 2 subunits of protein with molecular weights of 220-250 kd (5). Multiple isoforms of FN are produced from a single gene by alternative splicing at 3 distinct sites: EDA, EDB, and IIICS (5). Recent studies indicate that isoforms containing the EDA or EDB regions are expressed in early stages of fetal development, malignant transformation, and wound healing ( 5 ) , which suggests that EDAcontaining or EDB-containing FN may be expressed in association with cellular proliferation and transformation. The EDA-or EDB-containing FN is found in the synovial lining cells and small blood vessels of rheumatoid joints (6,7), and plasma levels of EDA+ FN in rheumatoid arthritis (RA) patients are higher than those in healthy individuals (8).We used immunohistochemistry techniques to show that EDA+ FN, which is specifically localized in the rheumatoid synovial lining layer, is synthesized by the synovial lining fibroblast-like (type B) cells. EDA+ FN is also detected in invasive fronts of active cellular pannus and on rheumatoid cartilage surfaces that are free of pannus. EDA+ FN is also colocalized with c-Fos protein in the rheumatoid synovial lining layer.Kazuo Hino, BS, Shunichi Shiozawa, MD, Yasuo Kuroki, MD, Hitoshi Ishikawa, MD, Kazuo Chihara, MD: Kobe University, Kobe, Japan; Kazuko Shiozawa, MD: Kakogawa National Hospital, Kakogawa, Japan; Kiyotoshi Sekiguchi, PhD: Osaka Medical Center for Maternal and Child Health, Osaka, Japan; Hisanobu Hirano, MS, Eiji Sakash...

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Human fibronectin: molecular cloning evidence for two mRNA species differing by an internal segment coding for a structural domain.

Cited by 219 publications

References 28 publications

Transformed human cells release different fibronectin variants than do normal cells.

Transformed human cells release different fibronectin variants than do normal cells.

Do neurons in the vertebrate CNS migrate on laminin?

Eda‐containing fibronectin is synthesized from rheumatoid synovial fibroblast‐like cells

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