An extracellular matrix-specific microarray allowed the identification of target genes downstream of discoidin domain receptors

Matrix Biol. 2003 Jun;22(4):373-81. doi: 10.1016/s0945-053x(03)00053-2.

Abstract

The two discoidin domain receptors, DDR1 and DDR2, are tyrosine kinases that are activated by collagen and are essential regulators of cell-matrix communication. However, the target genes downstream of activated DDRs and their physiological significance are largely unknown. Here, we describe a novel method to dissect signaling pathways induced by extracellular matrix (ECM) receptors. Using the doxycycline-inducible repression system (tet-off), we generated human fibrosarcoma and mouse fibroblast cell lines over-expressing DDR1 or DDR2. These cell lines were employed for gene expression analysis using microarrays specific for human and mouse genes coding for ECM proteins or ECM-interacting factors. We found that approximately 10% of the genes studied were up- or down-regulated more than twofold in response to signals generated by over-expressing DDRs. A common event downstream of DDR1 and DDR2 in human and mouse cells was the up-regulation of P-selectin glycoprotein ligand. Key target genes repressed upon DDR activation were agrin, syndecan-1 and alpha3 integrin. ECM-specific microarrays were found a valuable tool to dissect gene expression changes induced by collagen-receptor signaling pathways.

MeSH terms

  • Animals
  • Cell Line
  • Cluster Analysis
  • DNA, Complementary / genetics
  • Discoidin Domain Receptors
  • Extracellular Matrix / genetics*
  • Gene Expression Profiling*
  • Gene Expression Regulation*
  • Humans
  • Mice
  • Oligonucleotide Array Sequence Analysis*
  • Organ Specificity
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptor Protein-Tyrosine Kinases / deficiency
  • Receptor Protein-Tyrosine Kinases / genetics
  • Receptor Protein-Tyrosine Kinases / metabolism*
  • Receptors, Mitogen / deficiency
  • Receptors, Mitogen / genetics
  • Receptors, Mitogen / metabolism*

Substances

  • DNA, Complementary
  • RNA, Messenger
  • Receptors, Mitogen
  • Discoidin Domain Receptors
  • Receptor Protein-Tyrosine Kinases