Involvement of poly(ADP-ribose) polymerase 1 in ERBB2 expression in rheumatoid synovial cells.
Hyperplasia of synovial lining cells is one of the main features of rheumatoid arthritis (RA). We previously reported that ERBB2 is highly expressed in RA synovial cells and that it plays an important role in their hyperproliferative growth. Recent findings have suggested that poly(ADP-ribose) polymerase-1 (PARP-1) is involved in the transactivation of NF-kappaB-dependent genes such as ERBB2. In the present study, we investigated the role of PARP-1 in ERBB2 transcription in RA synovial cells. The expression level of PARP-1 was significantly high in synovial cells derived from three patients with RA, compared with three patients with osteoarthritis (OA). Luciferase assays revealed that PARP-1 augments the transcription of the ERBB2 gene and that a region between -404 and -368 is responsible for this activation. A protein with an apparent molecular mass of 115 kDa was isolated mainly from nuclear extracts of RA synovial cells with an affinity matrix harboring a DNA fragment identical to the above region. Mass spectrometric analysis demonstrated this protein to be PARP-1. Southwestern blot analysis showed that PARP-1 binds to this region, but not to adjacent regions. PARP-1 associates directly with NF-kappaB, and a chromatin immunoprecipitation assay indicated that these proteins interact with this enhancer region in the ERBB2 gene. Treatment of RA synovial cells with PARP-1 small interfering RNA attenuated their ERBB2 expression, while an inhibitor of the polymerase activity of PARP-1 had no effect. PARP-1 DNA binding is not required for transcriptional activation. These findings suggest that PARP-1 is involved in the expression of ERBB2 in concert with NF-kappaB, which might be associated with the proliferation of RA synovial cells.
Kitamura
Takuya
T
Department of Biomolecular Science and Orthopedics, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan.
Sekimata
Masayuki
M
Kikuchi
Shin-Ichi
S
Homma
Yoshimi
Y
D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000473:pathology; D001483:Base Sequence; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D006801:Humans; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D010003:Osteoarthritis / Q000378:metabolism / Q000473:pathology; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000502:physiology*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D013583:Synovial Membrane / Q000378:metabolism* / Q000473:pathology; D014158:Transcription, Genetic
D004247:DNA; D016328:NF-kappa B; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; D018719:Erb-b2 Receptor Tyrosine Kinases; C499320:PARP1 protein, human