作用于SIRT1和NAMPT的microRNAs调节HIV-1TAT的转录激活
发布时间：2018-10-29 19:26
【摘要】：人类免疫缺陷病毒（human immunodeficiency virus，HIV）是获得性免疫缺陷综合症（acquired immune deficiency syndrome，AIDS）的病原体，属于逆转录病毒。HIV-1长末重复端（long terminal repeat，LTR）上有一个调节基因tat编码反式激活蛋白（trans-activator of transcription，Tat）。Tat在HIV病毒复制过程中是必需的，它可以调节病毒基因组的mRNA转录的起始，并与细胞的凋亡有关。Tat在HIV-1基因表达的调控和生命周期中发挥重要作用，使其成为抗HIV治疗的新靶点。 本课题选择对HIV-1Tat基因起调节作用的沉默信息调节子2类似物1——也叫人类去乙酰化酶1（silent mating type information regulation2homolog1，SIRT1）作为靶点，选取与之相关的尼克酰胺磷酸核糖转移酶（nicotinamidephosphoribosyltransferase，Nampt）以及能与Nampt、SIRT1mRNA相作用的微小RNA（microRNAs）进行探索，检验是否在内源基因水平对Tat介导的HIV-1的转录与复制的调节起作用。 实验结果：通过实验发现一些microRNAs确实对Tat介导的HIV-1LTR的转录能够起到有效的抑制作用。其中miR-34、miR-199a、miR-217都能够通过对人类去乙酰化酶SIRT1的抑制作用对Tat介导的HIV-1的转录进行调控。与SIRT1紧密相关的尼克酰胺磷酸核糖转移酶Nampt能通过调节NAD~+水平调控SIRT1的活性，Nampt与NAD~+水平紧密相关。由于SIRT1是NAD~+依赖的，我们选取可能影响Nampt的miR-182进行实验，检验其对Tat介导的HIV-1转录的影响，实验结果发现miR-182能有效的降低Nampt的蛋白表达，同时由于Nampt与SIRT1的正相关性，，miR-182能有效的降低SIRT1mRNA及蛋白水平的表达，这些microRNAs都能有效地增强下游核因子NF-κB的p65亚基的乙酰化表达，p65亚基乙酰化的增加会促使转录的活化促进HIV-1的转录。而这些microRNA的抑制剂则能有效降低乙酰化p65的表达，使得HIV-1转录过程受到阻碍。实验表明通过寻找microRNAs作为抗HIV-1的新型治疗方式是可行的。
[Abstract]:Human immunodeficiency virus (human immunodeficiency virus,HIV) is the pathogen of acquired immunodeficiency syndrome (acquired immune deficiency syndrome,AIDS) and belongs to retrovirus. HIV-1 long terminal repeat (long terminal repeat, There is a regulatory gene tat encoding transactivator protein (trans-activator of transcription,Tat). Tat) on LTR that is necessary in the replication of the HIV virus, which regulates the initiation of mRNA transcription in the viral genome. Tat plays an important role in the regulation and life cycle of HIV-1 gene expression, making it a new target of anti-HIV therapy. In this study, we selected analogue 1 of silencing information regulator 2, also known as human deacetylase 1 (silent mating type information regulation2homolog1,SIRT1, which regulates the HIV-1Tat gene, as the target, and selected the related nicotinamidephosphoribosyltransferase, ribonucleosyltransferase (nicotinamidephosphoribosyltransferase,). Nampt) and tiny RNA (microRNAs) that interact with Nampt,SIRT1mRNA were explored to examine whether Tat mediated HIV-1 transcription and replication were regulated at the endogenous gene level. The results showed that some microRNAs could inhibit the transcription of HIV-1LTR mediated by Tat. MiR-34,miR-199a,miR-217 can regulate the transcription of HIV-1 mediated by Tat through the inhibition of human deacetylase SIRT1. Nicotinamide phosphate ribonucleosyltransferase (Nampt), closely related to SIRT1, can regulate the activity of SIRT1 by regulating the level of NAD~, and Nampt is closely related to the level of NAD~. Since SIRT1 is NAD~ dependent, we selected miR-182 that may affect Nampt to test its effect on HIV-1 transcription mediated by Tat. The results showed that miR-182 could effectively reduce the expression of Nampt protein. At the same time, because of the positive correlation between Nampt and SIRT1, miR-182 can effectively reduce the expression of SIRT1mRNA and protein. These microRNAs can effectively enhance the acetylation of p65 subunit of downstream nuclear factor NF- 魏 B. An increase in acetylation of p65 subunits promotes the activation of transcription to promote the transcription of HIV-1. These inhibitors of microRNA can effectively reduce the expression of acetylated p65 and block the process of HIV-1 transcription. The experiment shows that it is feasible to find microRNAs as a new type of anti-HIV-1 therapy.
【学位授予单位】：北京工业大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R373
[Abstract]:Human immunodeficiency virus (human immunodeficiency virus,HIV) is the pathogen of acquired immunodeficiency syndrome (acquired immune deficiency syndrome,AIDS) and belongs to retrovirus. HIV-1 long terminal repeat (long terminal repeat, There is a regulatory gene tat encoding transactivator protein (trans-activator of transcription,Tat). Tat) on LTR that is necessary in the replication of the HIV virus, which regulates the initiation of mRNA transcription in the viral genome. Tat plays an important role in the regulation and life cycle of HIV-1 gene expression, making it a new target of anti-HIV therapy. In this study, we selected analogue 1 of silencing information regulator 2, also known as human deacetylase 1 (silent mating type information regulation2homolog1,SIRT1, which regulates the HIV-1Tat gene, as the target, and selected the related nicotinamidephosphoribosyltransferase, ribonucleosyltransferase (nicotinamidephosphoribosyltransferase,). Nampt) and tiny RNA (microRNAs) that interact with Nampt,SIRT1mRNA were explored to examine whether Tat mediated HIV-1 transcription and replication were regulated at the endogenous gene level. The results showed that some microRNAs could inhibit the transcription of HIV-1LTR mediated by Tat. MiR-34,miR-199a,miR-217 can regulate the transcription of HIV-1 mediated by Tat through the inhibition of human deacetylase SIRT1. Nicotinamide phosphate ribonucleosyltransferase (Nampt), closely related to SIRT1, can regulate the activity of SIRT1 by regulating the level of NAD~, and Nampt is closely related to the level of NAD~. Since SIRT1 is NAD~ dependent, we selected miR-182 that may affect Nampt to test its effect on HIV-1 transcription mediated by Tat. The results showed that miR-182 could effectively reduce the expression of Nampt protein. At the same time, because of the positive correlation between Nampt and SIRT1, miR-182 can effectively reduce the expression of SIRT1mRNA and protein. These microRNAs can effectively enhance the acetylation of p65 subunit of downstream nuclear factor NF- 魏 B. An increase in acetylation of p65 subunits promotes the activation of transcription to promote the transcription of HIV-1. These inhibitors of microRNA can effectively reduce the expression of acetylated p65 and block the process of HIV-1 transcription. The experiment shows that it is feasible to find microRNAs as a new type of anti-HIV-1 therapy.
【学位授予单位】：北京工业大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R373
【参考文献】
相关期刊论文 前2条
1 孙晓娜;杨怡姝;曾毅;;反式激活应答元件RNA在HIV-1感染中的作用[J];中国生物化学与分子生物学报;2010年04期
2 张红胜;周s
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