苯中毒病人TOPOⅡα启动子调控因子Sp1组蛋白化学修饰改变

目的:研究拓扑异构酶Ⅱα(TOPOⅡα)启动子调控因子SP1组蛋白化学修饰在苯中毒病人中的改变。方法:25例临床慢性苯中毒患者骨髓单个核细胞为病例组,25例正常人骨髓单个核细胞为对照组,染色质免疫沉淀技术探讨TOPOⅡα启动子调控因子Sp1组蛋白乙酰化和甲基化水平的变化,RT-PCR法测定Sp1 mRNA的表达水平。结果:与对照组相比,临床苯中毒病例TOPOⅡα启动子调控因子Sp1组蛋白H4和H3乙酰化水平下降(P0.01),组蛋白H3K9甲基化水平升高(P0.01),组蛋白H3K4甲基化水平无明显改变。与正常对照组相比,临床苯中毒病例TOPOⅡα启动子调控因子Sp1的mRNA表达水平降低(P0.05)。结论:慢性苯中毒TOPOⅡα启动子调控因子Sp1组蛋白H4、H3乙酰化及H3K9甲基化修饰水平的改变伴随着mRNA水平的变化。TOPOⅡα启动子调控因子Sp1可能通过组蛋白H4、H3乙酰化及H3K9甲基化修饰改变在苯中毒所致的造血毒性中发挥作用。     

Methylation of histone H4 by arginine methyltransferase PRMT1 is essential in vivo for many subsequent histone modifications

PRMT1 is a histone methyltransferase that methylates Arg3 on histone H4. When we used siRNA to knock down PRMT1 in an erythroid cell line, it resulted in nearly complete loss of H4 Arg3 methylation across the chicken beta-globin domain, which we use as a model system for studying the relationship of gene activity to histone modification. We observed furthermore a domain-wide loss of histone acetylation on both histones H3 and H4, as well as an increase in H3 Lys9 and Lys27 methylation, both marks associated with inactive chromatin. To determine whether the effect on acetylation was directly related to the loss of H4 Arg3 methylation, we performed an in vitro acetylation reaction on chromatin isolated from PRMT1-depleted cells. We found that nucleosomes purified from these cells, and depleted in methylation at Arg3, are readily acetylated by nuclear extracts from the same cells, if and only if the nucleosomes are incubated with PRMT1 beforehand. Thus, methylation of histones by PRMT1 was sufficient to permit subsequent acetylation. Consistent with earlier reports of experiments in vitro, H4 Arg3 methylation by PRMT1 appears to be essential in vivo for the establishment or maintenance of a wide range of "active" chromatin modifications.     

HIV-1感染引起组蛋白乙酰化和增加p21WAF1表达

目的：DNA甲基化和组蛋白乙酰化是基因表达调控的主要形式。人类免疫缺陷病毒（HIV-1）可引起T淋巴细胞DNA甲基化酶上调。本文旨在明确HIV-1对细胞周期依赖刺激酶抑制剂p21^WAF1表达的影响。方法：建立HIV-1感染的Hut78细胞系；以RT-PCR和Westem blotting 分析p21^WAF1表达情况；以亚硫酸氢钠修饰DNA和基因测序，研究p21^WAF1基因启动子甲基化，以Western blot-ting 和染色体免疫测定探究总组蛋白和与p21^WAF1基因启动子相关的组蛋白乙酰化水平。并以GST pull-down和免疫沉淀分析HIV-1导致乙酰化及乙酰化引起p21^WAF1过表达的可能机理。结果：HIV-1感染后，其反式激活蛋白Tat与辅助转录因子P/CAF、hGCN5结合，共同刺激组蛋白H3乙酰化。尽管p21^WAF1启动子部分区域有甲基化发生，但p21^WAF1表达仍上调。这可能与E2A对p21^WAF1的作用有关。结论：HIV-1感染可引起T淋巴细胞p21^WAF1基因的甲基化和乙酰化紊乱，导致p21^WAF1表达增强。     

Epigenetic control of recombination in the immune system

Immune receptor gene expression is regulated by a series of developmental events that modify their accessibility in a locus, cell type, stage and allele-specific manner. This is carried out by a programmed combination of many different molecular mechanisms, including region-wide replication timing, changes in nuclear localization, chromatin contraction, histone modification, nucleosome positioning and DNA methylation. These modalities ultimately work by controlling steric interactions between receptor loci and the recombination machinery.     

组蛋白H3K9甲基化的多重作用

组蛋白是染色质的核心，其尾部共价修饰在基因表达调控中有重要作用。赖氨酸甲基化是组蛋白共价修饰之一，在多种生物学过程有重要作用。Suv39h1作为第一个被发现的组蛋白赖氨酸甲基转移酶，可以催化H3K9甲基化。H3K9甲基化是异染色质蛋白（HP1）染色区的停泊位点（docking site)。研究表明，组蛋白H3K9甲基化在异染色质形成及基因转录调控中具有重要的作用。