title: An extracellular matrix-specific microarray allowed the identification of target genes downstream of discoidin domain receptors.
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.
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ont, Canada M5S 1A8.
authors.forename: Elena
authors.initials: E
authors.lastname: Faraci
authors.forename: Maresa
authors.initials: M
authors.lastname: Eck
authors.forename: Bernhard
authors.initials: B
authors.lastname: Gerstmayer
authors.forename: Andreas
authors.initials: A
authors.lastname: Bosio
authors.forename: Wolfgang F
authors.initials: WF
authors.lastname: Vogel
mesh_terms: D000818:Animals; D002460:Cell Line; D016000:Cluster Analysis; D018076:DNA, Complementary / Q000235:genetics; D000070818:Discoidin Domain Receptors; D005109:Extracellular Matrix / Q000235:genetics*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D011975:Receptors, Mitogen; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases