小鼠巨噬細胞RAW264.7中hnRNP K磷酸化對COX-2 mRNA穩定度的調控

The regulatory role of hnRNP K phosphorylation to COX-2 mRNA stability in mouse macrophage RAW264.7 cells

江佩鈺(Pei-Yu Chiang)

指導教授 ： 張瀞仁

國立臺灣大學/生命科學院/生化科學研究所/博士(2012年)

https://doi.org/10.6342/NTU.2012.02372

摘要

當病原體感染發生在生物體中，許多細胞因子或即早基因，應迅速，準確地打開和關閉以進行免疫反應。嚴格地控制這些基因表現對預防免疫系統疾病是非常重要的。而利用RNA結合蛋白參與的轉錄後調控則是細胞用來操縱基因表達的策略之一。異質性核核糖核蛋白K，擁有三個可與RNA結合的KH（K-homology）結構域，並常涉及在調控後轉錄和轉譯的過程。為了研究hnRNP K在免疫反應中可能的功能，在脂多醣（LPS）處理過的小鼠RAW264.7巨噬細胞萃取物中以hnRNP K抗體進行RNA免疫沉澱。兩個即早基因：Tistetraproline（TTP）和腫瘤壞死因子-

關鍵字

脂多醣、巨噬細胞、RAW264.7 cell、異質性核核糖核蛋白K、環氧化酶2、轉錄後調控。

並列摘要

When pathogen infection occurrs in organism, many cytokines or immediate genes (IEGs) are rapidly and precisely expressed and/or repressed for performing immune responses. Tight control of these genes is very important for preventing immune disorders. Post-transcriptional regulation involving in trans-acting RNA-binding proteins is one of the favorite strategies of cells to manipulate gene expression. Heterogeneous nuclear ribonucleoprotein K (hnRNP K), containing three KH (K-homology) domains that mediate RNA binding, is involved in the regulation of post-transcription and translation processes. To study the possible functions of hnRNP K in immune response, I performed anti-hnRNP K RNA immunoprecipitation assay with cell extracts from lipopolysaccharide (LPS)-treated mouse RAW264.7 macrophages. Two IEGs, Tistetraproline (TTP) and Tumor necrosis factor

並列關鍵字

LPS, macrophage, RAW264.7, hnRNP K, COX-2, post-transcriptional regulation

參考文獻

1. Pinol-Roma S, Choi YD, Matunis MJ, Dreyfuss G (1988) Immunopurification of heterogeneous nuclear ribonucleoprotein particles reveals an assortment of RNA-binding proteins. Genes Dev 2: 215-227.

2. Choi YD, Dreyfuss G (1984) Isolation of the heterogeneous nuclear RNA-ribonucleoprotein complex (hnRNP): a unique supramolecular assembly. Proc Natl Acad Sci U S A 81: 7471-7475.

3. Dreyfuss G, Matunis MJ, Pinol-Roma S, Burd CG (1993) hnRNP proteins and the biogenesis of mRNA. Annu Rev Biochem 62: 289-321.

4. Dejgaard K, Leffers H, Rasmussen HH, Madsen P, Kruse TA, et al. (1994) Identification, molecular cloning, expression and chromosome mapping of a family of transformation upregulated hnRNP-K proteins derived by alternative splicing. J Mol Biol 236: 33-48.

5. Makeyev AV, Liebhaber SA (2002) The poly(C)-binding proteins: a multiplicity of functions and a search for mechanisms. RNA 8: 265-278.