The biological mechanism of Interferon Regulatory Factors

干扰素调节因子的生物学机制

• 批准号: 13672320
• 负责人: MASUMI Atsuko
• 金额: $ 2.3万
• 依托单位: National Institute of infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672320/
• 关键词: IRF; Acetylation; Chromatin; cell growth; Interferon; 転写因子; インターフェロン制御因子; 細胞分化・増殖

We have previously shown that interferon regulatory factor-2 (IRF-2) is acetylated by p300 and PCAF in vivo and in vitro. In this study we identified, by mass spectrometry, two lysine residues in the DNA binding domain (DBD), K75 and K78, to be the major acetylation sites in IRF-2. Although acetylation of IRF-2 did not alter DNA binding activity in vitro, mutation of K75 diminished IRF-2 dependent activation of histone H4 promoter activity. Acetylation of IRF-2 and IRF-2-stimulated H4 promoter activity were inhibited by the adenovirus E1A, indicating the involvement of p300/CBP. Mutation of K78, a residue conserved throughout the IRF family members, led to the abrogation of DNA binding activity independently of acetylation. H4 is transcribed only in rapidly growing cells and its promoter activity is dependent on cell growth. Consistent with a role for acetylated IRF-2 in cell growth control, IRF-2 was acetylated only in growing NIH 3T3 cells, but not in growth-arrested counterparts. Chromatin immunoprecipitation assays showed that IRF-2 interacted with p300 and bound to the endogenous H4 promoter only in growing cells, although the levels of total IRF-2 were comparable in both growing and growth-arrested cells. These results indicate that IRF-2 is acetylated in a cell growth-dependent manner, which enables it to contribute to transcription of cell growth regulated promoters.

我们之前已经证明，干扰素调节因子 2 (IRF-2) 在体内和体外均被 p300 和 PCAF 乙酰化。在这项研究中，我们通过质谱分析确定了 DNA 结合域 (DBD) 中的两个赖氨酸残基 K75 和 K78，它们是 IRF-2 中的主要乙酰化位点。尽管 IRF-2 的乙酰化不会改变体外 DNA 结合活性，但 K75 的突变减弱了组蛋白 H4 启动子活性的 IRF-2 依赖性激活。 IRF-2 的乙酰化和 IRF-2 刺激的 H4 启动子活性被腺病毒 E1A 抑制，表明 p300/CBP 的参与。 K78（IRF 家族成员中保守的残基）的突变导致 DNA 结合活性的废除，与乙酰化无关。 H4 仅在快速生长的细胞中转录，其启动子活性取决于细胞生长。与乙酰化 IRF-2 在细胞生长控制中的作用一致，IRF-2 仅在生长的 NIH 3T3 细胞中被乙酰化，而在生长停滞的细胞中则不然。染色质免疫沉淀测定表明，IRF-2 仅在生长细胞中与 p300 相互作用并与内源 H4 启动子结合，尽管总 IRF-2 水平在生长细胞和生长停滞细胞中相当。这些结果表明IRF-2以细胞生长依赖性方式被乙酰化，这使其能够促进细胞生长调节启动子的转录。

项目成果

Masumi A., Ozato K.: "Coactivator p300 acetylates the interferon ragulatory factor-2 in U937 cells following phorbol ester treatment"J.Biol.Chem.. 276. 20973-20980 (2001)

Masumi A.、Ozato K.：“佛波酯处理后，辅激活剂 p300 乙酰化 U937 细胞中的干扰素调控因子 2”J.Biol.Chem.. 276. 20973-20980 (2001)

Caillaud A., Prakash A., Smith E., Masumi A., Hovanessian A.G., Levy D.E., Marie I: "Acetylation of Interferon Ragulatory Factor-7 by p300/CREB-binding Protein (CBP)-associated factor (PCAF) Impairs its DNA binding"J.Biol.Chem.. 277. 49417-49421 (2002)

Caillaud A.、Prakash A.、Smith E.、Masumi A.、Hovanessian A.G.、Levy D.E.、Marie I：“p300/CREB ​​结合蛋白 (CBP) 相关因子 (PCAF) 乙酰化干扰素调节因子 7 会损害

Masumi A, Tamaoki S, Wang IM, Ozato K, and Komuro K: "IRF-8/ICSBP and IRF-1 cooperatively stimulate mouse IL-12 promoter activity in macrophages"FEES Lett.. 531. 348-353 (2002)

Masumi A、Tamaoki S、Wang IM、Ozato K 和 Komuro K：“IRF-8/ICSBP 和 IRF-1 协同刺激巨噬细胞中的小鼠 IL-12 启动子活性”FEES Lett.. 531. 348-353 (2002)

Caillaud A., Prakash A., Smith E., Masumi A., Hovanessian A.G., Levy D.E. and Marie I.: "Acetylation of Interferon Regulatory Factor-7 by p300/CREB-binding Protein (CBP)-associated factor (PCAF) Impairs its DNA binding"J. Biol. Chem.. 277. 49417-49421 (20

Caillaud A.、Prakash A.、Smith E.、Masumi A.、Hovanessian A.G.、Levy D.E.

Atsuko Masumi, Keiko Ozato: "Conctivator p300 A cetylates the Interferon Regulatory Factor-2 in V937 cells follawing Phorbol Ester treatment"J.Boil.Chem.. 276. 20973-20980 (2001)

Atsuko Masumi、Keiko Ozato：“Conttivator p300 A 在佛波醇酯处理后对 V937 细胞中的干扰素调节因子 2 进行乙酰化”J.Boil.Chem.. 276. 20973-20980 (2001)