Isolation of cDNA clones specific for collagen IV and laminin from mouse teratocarcinoma cells.

Wang, Sho‐Ya; Gudas, Lorraine J.

doi:10.1073/pnas.80.19.5880

Cited by 142 publications

(61 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For Northern blot analysis, total RNA was extracted using guanidine isothiocyanate-CsCl 2 density gradient centrifugation as previously described (26). The total RNA was size fractionated and transferred to a nylon membrane (GeneScreen Plus, DuPont, Boston, MA) and hybridized to a 32 P-labeled (random-primer labeling kit, BMB, Indianapolis, IN) 1.4-kilobase pair human type VII collagen cDNA probe containing NC2/COL domain.…”

Section: Methodsmentioning

confidence: 99%

The Carboxyl Terminus of Type VII Collagen Mediates Antiparallel Dimer Formation and Constitutes a New Antigenic Epitope for Epidermolysis Bullosa Acquisita Autoantibodies

Chen¹,

Keene

Costa

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

Type VII collagen, the major component of anchoring fibrils, consists of a central collagenous triple-helical domain flanked by two noncollagenous domains, NC1 and NC2. The NC2 domain has been implicated in catalyzing the antiparallel dimer formation of type VII procollagen. In this study, we produced the entire 161 amino acids of the NC2 domain plus 186 amino acids of adjacent collagenous domain (NC2/COL) and purified large quantities of the recombinant NC2/COL protein. Recombinant NC2/ COL readily formed disulfide-bonded hexamers, each representing one antiparallel dimer of collagen VII. Removal of the collagenous helical domain from NC2/COL by collagenase digestion abolished the antiparallel dimer formation. Using site-directed mutagenesis, we found that mutation of either cysteine 2802 or cysteine 2804 alone within the NC2 domain blocked antiparallel dimer formation. In contrast, a single cysteine mutation, 2634, within the collagenous helical domain had no effect. A generated methionine to lysine substitution, M2798K, that is associated with recessive dystrophic epidermolysis bullosa, was unable to form antiparallel dimers. Furthermore, autoantibodies from epidermolysis bullosa acquisita patients also reacted with NC2/COL. We conclude that NC2 and its adjacent collagenous segment mediate antiparallel dimer formation of collagen VII. Epidermolysis bullosa acquisita autoantibodies bound to this domain may destabilize anchoring fibrils by interfering with antiparallel dimer assembly leading to epidermal-dermal disadherence.Type VII collagen, a genetically distinct member of the collagen family, is found within the basement membrane zone beneath stratified squamous epithelium (1, 2). Type VII collagen is a major component of anchoring fibrils, attachment structures within the basement membrane between the epidermis and dermis of human skin (3, 4). In inherited forms of dystrophic epidermolysis bullosa (DEB), 1 anchoring fibrils are diminutive and/or reduced in number (5-7). In addition to inherited DEB, epidermolysis bullosa acquisita (EBA) is an acquired autoimmune form of epidermolysis bullosa. EBA is characterized by circulating and tissue-bound IgG autoantibodies to type VII collagen (8, 9). Like DEB, ultrastructural studies have demonstrated a dramatic paucity of anchoring fibrils in EBA skin (10). These observations suggest that type VII collagen plays an important role in maintaining epidermal-dermal adherence. Type VII collagen has been cloned, and a genetic linkage has been established between inherited DEB and mutations in the gene that encodes for type VII collagen, COL7A1 (11-14). There have been over 100 distinct COL7A1 gene mutations identified in patients with DEB, and these mutations have occurred within NC1, NC2, and the helical domain (15,16).Type VII collagen is composed of three identical ␣ chains, each consisting of a 145-kDa central collagenous triple-helical segment, flanked by a large 145-kDa amino-terminal non-collagenous domain (NC1), and a smaller 34-kDa carboxyl-terminal non-collagen...

show abstract

Section: Methodsmentioning

confidence: 99%

The Carboxyl Terminus of Type VII Collagen Mediates Antiparallel Dimer Formation and Constitutes a New Antigenic Epitope for Epidermolysis Bullosa Acquisita Autoantibodies

Chen¹,

Keene

Costa

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Because overexpression of the SAC RAR-CB vector yields cells that appear to be undifferentiated EC cells, studies were performed to compare the expression of some retinoic acid-responsive genes in these clones compared with normal F9 EC cells. Endo B (Singer et al 1986), t-PA (Rickles et al 1988), B-1 laminin (Wang and Gudas 1983), and type IV collagen [alpha-2 (Wang and Gudas 1983); alpha-1 (Rickles et al 1988)] are transcriptionally induced to high levels of expression within 48-72 hr after addition of retinoic acid plus dibutyryl cAMP to F9 cells. Conversely, the level of c-myc mRNA decreases dramatically within 8 hr of retinoic acid administration to F9 cells (Campisi et al 1984;Dony et al 1985).…”

Section: >Cb Sac Rar-cbmentioning

confidence: 99%

Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells.

Espeseth¹,

Murphy²,

Linney³

1989

Genes Dev.

View full text Add to dashboard Cite

“…ment of F9 Cells-To determine the temporal relationship between induction of the ␣1,3GT compared by RA to other known markers and to assess the efficiency of differentiation of F9 cells in response to 10 Ϫ7 M RA in our system, we determined the amount of laminin secreted into the cell culture media. It has been shown that the steady-state levels of mRNAs for laminin B1 and B2 increase dramatically following differentiation of F9 cells, and laminin has been defined as a differentiation marker, along with other basement membrane components, such as type IV collagen (␣1) (39,(55)(56)(57)(58)(59)(60). Laminin was detected by an enzyme-linked immunosorbent assay using cell culture media immobilized in microtiter wells and anti-laminin antibody as the detecting reagent.…”

Section: Time Course Of Induction Of ␣13gt Enzyme Activity In F9 Celmentioning

confidence: 99%

Transcriptional Regulation of α1,3-Galactosyltransferase in Embryonal Carcinoma Cells by Retinoic Acid

Cho

Yeh

Cho

et al. 1996

Journal of Biological Chemistry

View full text Add to dashboard Cite

Treatment of mouse teratocarcinoma F9 cells with alltrans-retinoic acid (RA) causes a 9-fold increase in steady-state levels of mRNA for UDP-Gal:␤-D-Gal ␣1,3-galactosyltransferase (␣1,3GT) beginning at 36 h. Enzyme activity rises in a similar fashion, which also parallels the induction of laminin and type IV collagen. Nuclear run-on assays indicate that this increase in ␣1,3GT in RA-treated F9 cells, like that of type IV collagen, is transcriptionally regulated. Differentiation also results in increased secretion of soluble ␣1,3GT activity into the growth media. The major ␣-galactosylated glycoprotein present in the media of RA-treated F9 cells, but not of untreated cells, was identified as laminin. Differentiation of F9 cells is accompanied by an increase in ␣-galactosylation of membrane glycoproteins and a decrease in expression of the stage-specific embryonic antigen, SSEA-1 (also known as the Lewis X antigen or Le X ), which has the structure Gal␤1-4(Fuc␣1-3)GlcNAc␤1-R. However, flow cytometric analyses with specific antibodies and lectins, following treatment of cells with ␣-galactosidase, demonstrate that differentiated cells contain Le X antigens that are masked by ␣-galactosylation. Thus, RA induces ␣1,3GT at the transcriptional level, resulting in major alterations in the surface phenotype of the cells and masking of Le X antigens.

show abstract

Isolation of cDNA clones specific for collagen IV and laminin from mouse teratocarcinoma cells.

Cited by 142 publications

References 35 publications

The Carboxyl Terminus of Type VII Collagen Mediates Antiparallel Dimer Formation and Constitutes a New Antigenic Epitope for Epidermolysis Bullosa Acquisita Autoantibodies

The Carboxyl Terminus of Type VII Collagen Mediates Antiparallel Dimer Formation and Constitutes a New Antigenic Epitope for Epidermolysis Bullosa Acquisita Autoantibodies

Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells.

Transcriptional Regulation of α1,3-Galactosyltransferase in Embryonal Carcinoma Cells by Retinoic Acid

Contact Info

Product

Resources

About