Collagenase is a major gene product of induced rabbit synovial fibroblasts

Journal Article · Tue May 01 04:00:00 EDT 1984 · J. Cell Biol.; (United States)

DOI:https://doi.org/10.1083/jcb.98.5.1656· OSTI ID:5807833

Aggeler, J; Frisch, S M; Werb, Z

The effects of the tumor-promoting phorbol diester, 12-0-tetradecanoylphorbol-13-acetate (TPA), on rabbit synovial fibroblasts was investigated. This agent induced a major switch in gene expression in these cells that was marked by the specific induction of the neutral proteinase, collagenase, and was always accompanied by alteration in cell morphology. Procollagenase synthesis and secretion was first observed 6-12 h after the addition of TPA. The rate of collagenase production depended on the TPA concentration (1-400 ng/ml) and time of exposure (1-72 h). Procollagenase was the most prominent protein visible by direct silver straining or by autoradiography after SDS PAGE of (/sup 35/S)methionine-labeled proteins. The two procollagenase bands of M/sub r/ 53,000 and 57,000, which migrated as a family of spots on two-dimensional gels and were immunoprecipitated by antibodies to purified rabbit collagenase, accounted for 23% of the newly synthesized, secreted protein in TPA-treated cells. Cell-free translation of mRNA from TPA-treated cells in rabbit reticulocyte lysate produced a single band of immunoprecipitable preprocollagenase as a major product (5% of total) that migrated as a single spot on two-dimensional gels. Secreted procollagenase preprocollagenase, and active collagenase (purified to homogeneity; specific activity 1.2 X 10/sup 4/ U/mg protein) had related peptide maps. Two other major secreted proteins, a neutral metalloproteinase of M/sur r/ 51,000 and a polypeptide of M/sub r/ 47,000, were also induced by TPA. In contrast to the induction of these four polypeptides, TPA decreased synthesis and secretion of a number of proteins, including collagen and fibronectin. Collagenase is a convenient marker for major alterations in the pattern of protein synthesis and secretion of rabbit synovial fibroblasts treated with TPA. 50 references, 5 figures.

• Research Organization: Univ. of California, San Francisco
• DOE Contract Number: AC03-76SF01012
• OSTI ID: 5807833
• Journal Information: J. Cell Biol.; (United States), Journal Name: J. Cell Biol.; (United States) Vol. 98:5; ISSN JCLBA
• Country of Publication: United States
• Language: English

Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL EFFECTS
BIOSYNTHESIS
CARBOXYLIC ACIDS
CARCINOGENS
CHEMISTRY
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DRUGS
ENZYME ACTIVITY
ENZYMES
ESTERS
EVEN-ODD NUCLEI
FIBROBLASTS
GENES
GENETIC EFFECTS
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MESSENGER-RNA
METHIONINE
NUCLEI
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDE HYDROLASES
PEPTIDES
PHORBOL ESTERS
POLYPEPTIDES
PROTEINS
RABBITS
RADIOISOTOPES
RNA
SOMATIC CELLS
SULFUR 35
SULFUR ISOTOPES
SYNTHESIS
TRACER TECHNIQUES
VERTEBRATES