I. CPAPAP-1 蛋白質在 NF-κB 途徑上功能性之研究II. 傳統中醫藥對樹突細胞活性上的抑制效應

Abstract

I. The functional study of CPAPAP-1 protein in NF-κB pathway. Nuclear factor-κB (NF-κB) is a Rel transcription factor that controls the gene expression involved in immune response, inflammation, apoptosis, and proliferation. In mammals, five Rel members including NF-κB1 (p50/p105), NF-κB2 (p52/p100), RelA/p65, RelB, and c-Rel have been identified. Koyanagi et al. performed a yeast two-hybrid screening using the N-terminal domain of the RelA as bait and obtained centrosomal P4.1-associated protein (CPAP) as a candidate of associating partner. Molecular biology assays and cell biology experiments showed that the role of CPAP is a coactivator of NF-κB-mediated transcription. In addition, CPAP was also revealed as a coactivator of Stat5-mediated transcription and a component of the centrosomal complex regulated centriole length. The mutation of CPAP resulted in Autosomal recessive primary microcephaly (MCPH). The C-terminal region of CPAP appears to contribute NF-κB-mediated transcriptional activation. However, knowledge about the factors that interact with the C-terminal region of CPAP is still little. In a yeast two-hybrid screen using the C-terminal region of CPAP as bait, we identified a novel associating partner named CPAP-associated protein-1 (CPAPAP-1, GenBank: BC114945). We also confirmed that CPAPAP-1 associates with N-terminal region of RelA and has the effector domain of the CAP family of transcription factors. In most cases, the effector (cAMP) binding the domain induced conformational changes and activated transcription. A portion of CPAPAP-1 was observed to accumulate in the nucleus, although CPAPAP-1 was found primarily in the cytoplasm without any stimulation. In the future, we will confirm that CPAPAP-1 is a coactivator of NF-κB-mediated transcription by using NF-κB-dependent reporter gene expression assay and RNA interference technique. Furthermore, we will assess physiological function of CPAPAP-1 through the knockout mouse model of CPAPAP-1. II. The inhibitory effect of Tranditional Chinese medicine on activation of dendritic cell Traditional Chinese medicine (TCM) has been used for a long history for the treatment in human disease. Many reports showed TCM clinic curative effects due to regulation of immune response. Dendritic cells (DCs) called antigen presenting cell that specialize in the uptake and presentation of antigens to effector cells. DCs can be a therapeutic target to cure cancer, parasites and autoimmune disease by enhancement or inhibition activation and function of DCs. In this study, we identified candidates of Traditional Chinese medicine (TCM) that attenuate Dendritic cell (DCs) activation from natural source. We treated mouse dendritic cell line (DC2.4) with TCM for 24 hours and then stimulated cells with LPS to analyze cytokine secretion, expression of co-stimulatory molecule and ability of antigen uptake. Finally, we obtained some TCM having ability to inhibit activation of DCs. In the future, more mechanistic studies will be needed to elucidate the molecular basis of inhibition effect of TCM.