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PMID: 9015316 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A three-dimensional collagen lattice induces protein kinase C-zeta activity: role in alpha2 integrin and collagenase mRNA expression.

The Journal of cell biology ·Vol. 136 ·No. 2 ·1997-01-27 ·Pages 473-83

Xu J, Clark RA

Abstract

A three-dimensional collagen lattice can provide skin fibroblasts with a cell culture environment that simulates normal dermis. Such a collagen matrix environment regulates interstitial collagenase (type I metalloproteinase [MMP-1], collagenase-1) and collagen receptor alpha2 subunit mRNA expression in both unstimulated or platelet-derived growth factor-stimulated dermal fibroblasts (Xu, J., and R.A.F. Clark. 1996. J. Cell Biol. 132:239-249). Here we report that the collagen gel can signal protein kinase C (PKC)-zeta activation in human dermal fibroblasts. An in vitro kinase assay demonstrated that autophosphorylation of PKC-zeta immunoprecipitates was markedly increased by a collagen matrix. In contrast, no alteration in PKC-zeta protein levels or intracellular location was observed. DNA binding activity of nuclear factor kappaB (NF-kappaB), a downstream regulatory target of PKC-zeta, was also increased by fibroblasts grown in collagen gel. The composition of the NF-kappaB/Rel complexes that contained p50, was not changed. The potential role of PKC-zeta in collagen gel-induced mRNA expression of collagen receptor alpha2 subunit and human fibroblast MMP-1 was assessed by the following evidence. Increased levels of alpha2 and MMP-1 mRNA in collagen gel-stimulated fibroblasts were abrogated by bisindolylmaleimide GF 109203X and calphostin C, chemical inhibitors for PKC, but retained when cells were depleted of 12-myristate 13-acetate (PMA)-inducible PKC isoforms by 24 h of pretreatment with phorbol PMA. Antisense oligonucleotides complementary to the 5' end of PKC-zeta mRNA sequences significantly reduced the collagen lattice-stimulated alpha2 and MMP-1 mRNA levels. Taken together, these data indicate that PKC-zeta, a PKC isoform not inducible by PMA or diacylglycerol, is a component of collagen matrix stimulatory pathway for alpha2 and MMP-1 mRNA expression. Thus, a three-dimensional collagen lattice maintains the dermal fibroblast phenotype, in part, through the activation of PKC-zeta.

MeSH Terms
Antigens, CD/biosynthesis,genetics Becaplermin Cell Nucleus/enzymology Cells, Cultured Collagen/chemistry,physiology Collagenases/biosynthesis,genetics Cytoplasm/enzymology DNA/metabolism Enzyme Activation Gels Humans Integrin alpha2 Matrix Metalloproteinase 1 NF-kappa B/metabolism Oligonucleotides, Antisense Phosphorylation Platelet-Derived Growth Factor/pharmacology Protein Biosynthesis Protein Kinase C/metabolism Proto-Oncogene Proteins c-sis RNA, Messenger/genetics,metabolism Tetradecanoylphorbol Acetate/pharmacology Thionucleotides
Chemicals
Antigens, CD Gels Integrin alpha2 NF-kappa B Oligonucleotides, Antisense Platelet-Derived Growth Factor Proto-Oncogene Proteins c-sis RNA, Messenger Thionucleotides Becaplermin Collagen DNA protein kinase C zeta Protein Kinase C Collagenases Matrix Metalloproteinase 1 Tetradecanoylphorbol Acetate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Xu J
Department of Dermatology, School of Medicine, SUNY at Stony Brook, New York 11794-8165, USA.
Clark R A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.136.2.473
Published
1997-01-27
Pages
473-83
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2134818
Subset
IM
Grants
NIA NIH HHS · AG10114309 · United States
External Links
PubMed source DOI: 10.1083/jcb.136.2.473 PMC Full Text View journal details
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