- Poster presentation
- Open access
- Published:
GCIP, Id like HLH protein, negatively regulates cell proliferation of rheumatoid synovial cells via interaction with CBP
Arthritis Research & Therapy volume 14, Article number: P17 (2012)
Background
Rheumatoid arthritis (RA) is one of the most common articular diseases with a prevalence of 1% worldwide [1, 2]. The clinical features of RA include chronic inflammation of systemic joints associated with synovial hyperplasia followed by impairment of quality of life [3, 4]. Recently, we have shown that Synoviolin/Hrd1, an E3 ubiquitin ligase, is a novel causative factor for arthropathy [5]. However, the mechanism that regulates synovial cell outgrowth is not fully understood.
Materials and methods
Human embryonic kidney (HEK)-293 cells, HEK-293T cells, NIH3T3 cells and synovial cells were cultured in DMEM medium. Transient transfection assays were performed in HEK-293 cells and HEK-293T cells. HEK-293 cells transfected with NF-κB-Luc were treated with 100 ng/ml of phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), or 10 ng/ml of TNF-α for 24 h, and luciferase activities were measured. siRNAs with 21 nucleotides for human GCIP were chemically synthesized. Transfection with siRNAs and cell survival assay were carried out.
Results
Grap2 cyclin D interacting protein (GCIP), Id like HLH protein, was down-regulated in the rheumatoid synovial cells. Introduction of GCIP into mouse fibroblast NIH3T3 cells resulted in growth suppression, whereas knockdown with siRNAs in synovial cells enhanced cell growth. GCIP associated with CBP and repressed transcription of CREB-target genes such as cyclin D1 by inhibition of interaction between CBP and RNA polymerase II complexes. Binding assays revealed that GCIP bound to CBP via acidic region, not HLH domain, and this interaction was regulated by phosphorylation of GCIP in a cell cycle-dependent manner. Therefore, GCIP has inhibitory effect on cell proliferation via interference with CBP-mediated transcription.
Conclusions
We propose the novel inhibitory mechanisms of Id protein family; the coactivator CBP is a functional target. Furthermore, down-regulation of GCIP may be a key factor in rheumatoid synovial cell outgrowth.
References
Harris ED: Rheumatoid arthritis. Pathophysiology and implications for therapy. N Engl J Med. 1990, 322: 1277-1289. 10.1056/NEJM199005033221805.
Feldmann M, Brennan FM, Maini RN: Rheumatoid arthritis. Cell. 1996, 85: 307-310. 10.1016/S0092-8674(00)81109-5.
Aarvak T, Natvig JB: Cell-cell interactions in synovitis: antigen presenting cells and T cell interaction in rheumatoid arthritis. Arthritis Res. 2001, 3: 13-17. 10.1186/ar135.
Schett G, Tohidast-Akra M, Steiner G, Smolen J: The stressed synovium. Arthritis Res. 2001, 3: 80-86. 10.1186/ar144.
Amano T, Yamasaki S, Yagishita N, Tsuchimochi K, Shin H, Kawahara K, Aratani S, Fujita H, Zhang L, Ikeda R, Fujii R, Miura N, Komiya S, Nishioka K, Maruyama I, Fukamizu A, Nakajima T: Synoviolin/Hrd1, an E3 ubiquitin ligase, as a novel pathogenic factor for arthropathy. Genes Dev. 2003, 17: 2436-2449. 10.1101/gad.1096603.
Workman CJ, Wang Y, El Kasmi KC, Pardoll DM, Murray PJ, Drake CG, Vignali DA: LAG-3 regulates plasmacytoid dendritic cell homeostasis. J Immunol. 2009, 182: 1885-1891. 10.4049/jimmunol.0800185.
Treibel F, Hacene K, Pichon MF: A soluble lymphocyte activation gene-3 (sLAG-3) protein as a prognostic factor in human breast cancer expressing estrogen or progesterone receptors. Cancer Letters. 2006, 235: 147-153. 10.1016/j.canlet.2005.04.015.
Triebel F: LAG-3: a regulator of T-cell and DC responses and its use in therapeutic vaccination. Trends Immunol. 2003, 24: 619-622. 10.1016/j.it.2003.10.001.
Author information
Authors and Affiliations
Rights and permissions
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
About this article
Cite this article
Fujita, H., Nakazawa, M., Aratani, S. et al. GCIP, Id like HLH protein, negatively regulates cell proliferation of rheumatoid synovial cells via interaction with CBP. Arthritis Res Ther 14 (Suppl 1), P17 (2012). https://doi.org/10.1186/ar3618
Published:
DOI: https://doi.org/10.1186/ar3618