Anti-UBLCP1 monoclonal antibody, clone FQS22034

RNA polymerase II C-terminal domain phosphatase 1 containing ubiquitin like domain (UBLCP1) has been identified as a regulatory molecule of RNA polymerase II. UBLCP1 consists of ubiquitin like domain (UBL) and phosphatase domain homologous with UDP and CTD phosphatase. UBLCP1 was cloned into the E. coli express ion vectors, pET32a and pGEX 4T-1 with TEV protease cleavage site and purified using both affinity and gel-filtration chromatography. Domains of UBLCP1 protein were successfully purified as 7 mg/500 mL (UBLCP1, 36.78 KDa), 32 mg/500 mL (UBL, 9 KDa) and 8 mg/500 mL (phosphatase domain, 25 KDa) yielded in LB medium, respectively. Isotope-labeled samples including triple-labeled (H-2/N-15/C-13) UBLCP1 were also prepared for hetero-nuclear NMR experiments. N-15-H-1 2D-HSQC spectra of UBLCP1 suggest that both UBL and phosphatase domain are properly folded and structurally independent each other. These data will promise us further structural investigation of UBLCP1 by NMR spectroscopy and/or X-ray crystallography.

PURPOSE. Long noncoding RNAs (lncRNAs) are emerging as important regulators of inflammatory immune responses, whereby genetic variants may affect this biologic function. This study aimed to investigate the association of 110 single nucleotide polymorphisms (SNPs) of lncRNAs, known to be associated with autoimmune disease, in patients with ocular Vogt-Koyanagi-Harada (VKH) disease, Behcet's disease (BD), and acute anterior uveitis (AAU) with or without ankylosing spondylitis (AS). METHODS. A two-stage case-control study was performed on 1626 VKH patients, 384 BD patients, 624 AAU with AS, 751 AAU without AS, 720 AS without AAU, and 3305 healthy subjects. lncRNAs 110 SNPs were genotyped using MassARRAY System or TaqMan SNP assays. The gene expression and cytokine production were measured using real-time PCR or ELISA. RESULTS. The frequency of the C allele of rs4937362 in RP11-264E20.1 was markedly decreased in the AS without AAU group compared with controls (Combined P = 9.37 x 10(-7), odds ratio [OR] = 0.73). An increased frequency of the A allele of rs6871626 between UBLCP1, IL12B, and LOC285627 was found in VKH patients compared with controls (Combined P = 1.88 3 10(-4), OR = 1.19). UBLCP1, IL12B, and LOC285627 were expressed in human uveal tissues. Functional studies showed a decreased LOC285627 mRNA expression in peripheral blood mononuclear cells (PBMCs) and an increased IL-10 production in PBMCs following LPS stimulation in rs6871626 CC genotype carriers. CONCLUSIONS. Our study is the first to show that rs4937362/RP11-264E20.1 is associated with AS and that rs6871626 is associated with VKH disease in Chinese Han. The protective rs6871626 genotype was shown to regulate the expression of LOC285627 and to increase the production of the anti-inflammatory cytokine IL-10.