Lysyl oxidase activates the transcription activity of human collagene III promoter. Possible involvement of Ku antigen.
Lysyl oxidase is an extracellular enzyme that controls the maturation of collagen and elastin. Lysyl oxidase and collagen III often show similar expression patterns in fibrotic tissues. Therefore, we investigated the influence of lysyl oxidase overexpression on the promoter activity of human COL3A1 gene. Our results showed that when COS-7 cells overexpressed the mature form of lysyl oxidase, the activity of the human COL3A1 promoter was increased up to an average of 12 times when tested by luciferase reporter assay. The effect was specific, because other promoters were not affected. Moreover, lysyl oxidase effect was abolished by beta-aminopropionitrile, a specific inhibitor of its catalytic activity. Electrophoretic mobility shift assay showed a binding activity in the region from -101 to -77 that was significantly increased by lysyl oxidase overexpression. The binding was specifically competed by the cold probe, and the mutagenesis of this region abolished both the binding activity in gel retardation and lysyl oxidase stimulation of COL3A1 promoter in transfection experiments. We identified the binding activity as Ku antigen in its two components: Ku80 and Ku70. This study suggests a new coordinated mechanism by which lysyl oxidase might control the development of fibrosis.
Giampuzzi
M
M
Department of Nephrology, Gaslini Children's Hospital, Genova, Italy.
Botti
G
G
Di Duca
M
M
Arata
L
L
Ghiggeri
G
G
Gusmano
R
R
Ravazzolo
R
R
Di Donato
A
A
D000629:Aminopropionitrile / Q000494:pharmacology; D000818:Animals; D034961:Antigens, Nuclear; D001483:Base Sequence; D019556:COS Cells; D003094:Collagen / Q000235:genetics*; D004265:DNA Helicases; D015342:DNA Probes / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D005347:Fibroblasts; D005355:Fibrosis / Q000201:enzymology / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D000072200:Ku Autoantigen; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D008249:Protein-Lysine 6-Oxidase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D015533:Transcriptional Activation; D014162:Transfection
D034961:Antigens, Nuclear; D015342:DNA Probes; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D000629:Aminopropionitrile; D003094:Collagen; D008249:Protein-Lysine 6-Oxidase; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen