Cell-matrix interactions induce tyrosine phosphorylation of MAP kinases ERK1 and ERK2 and PLCgamma-1 in two-dimensional and three-dimensional cultures of human fibroblasts.

Using immunoprecipitation and phosphotyrosine detection by Western blotting, intracellular signaling intermediates were analyzed in human primary dermal fibroblasts, either seeded as monolayers on collagen I coats (2D) or seeded within three-dimensional collagen I lattices (3D). Previous results demonstrated that integrin activation in these systems resulted in a cascade of protein tyrosine phosphorylation, including focal adhesion kinase (D. Roeckel and T.

Three-dimensional contact with type I collagen mediates tyrosine phosphorylation in primary human fibroblasts.

By applying Western blot analysis using anti-phosphotyrosine antibodies, primary human dermal fibroblasts were examined after having been cultured on type I collagen-coated surfaces or in free-floating type I collagen gels. In both systems cells showed enhanced tyrosine phosphorylation of a M(r) 120,000 protein (pp120) and of a M(r) 42,000 protein (pp42). Phosphorylation was apparent 6 h at the latest after initiation of the culture and was only slightly induced on polylysine or on plastic.

Identification of binding sites on the regulatory A subunit of protein phosphatase 2A for the catalytic C subunit and for tumor antigens of simian virus 40 and polyomavirus.

Protein phosphatase 2A is composed of three subunits: the catalytic subunit C and two regulatory subunits, A and B. The A subunit consists of 15 nonidentical repeats and has a rodlike shape. It is associated with the B and C subunits as well as with the simian virus 40 small T, polyomavirus small T, and polyomavirus medium T tumor antigens.

Expression of an early Epstein-Barr virus antigen (EA-D) in E. coli. Brief report.

The 1.34 kb BcII-BgIII-fragment of the BamHI-M region of Epstein-Barr virus genome, comprising the complete BMRF1 open reading frame, was cloned into the tryptophan regulated E. coli expression vector pATH1.

Biochemical characterization of two Epstein-Barr virus early antigen-associated phosphopolypeptides.

Epstein-Barr virus (EBV) nonproducer cells NC37 induced to viral early antigen (EA) synthesis by the tumor promotor 12-O-tetradecanoylphorbol-13-acetate (TPA) were labeled at Day 4 after induction with 32P, and were analyzed by immunoprecipitation with human EA-positive sera. By employing this method the appearance of two phosphopolypeptides of 50 and 58K (pp50 and pp58) was well correlated with EA complex. Partial V8 protease digestion and two-dimensional peptide analysis revealed that the polypeptides pp50 and pp58 are related.