二磷酸腺苷核糖多聚酶的研究进展

二磷酸腺苷核糖多聚酶[Poly（ADP-Ribose）Polymerase,PAPe]是一类具有重要生理功能的蛋白酶。PARP能催化二磷酸腺苷核糖多聚化反应。二磷酸腺苷核糖多聚化对DNA修复和基因组稳定性起着重要作用。但PARP的过激活与许多疾病的病理机制有关。介绍了PARP的结构和功能,PARP家族的同族体以及PARP在一些疾病病理机制中的作用。     

聚腺苷二磷酸-核糖聚合酶1与DNA双链断裂修复的相关性

DNA双链断裂(double strand breaks,DSBs)是细胞最严重的DNA损伤形式。细胞通过同源重组(homologous recombination,HR)和非同源末端连接(non-homologous end joining,NHEJ)途径修复DNA双链断裂损伤。聚腺苷二磷酸核糖基化(poly(ADP-ribosyl)ation,PARylation)是蛋白质翻译后修饰过程,这个过程由聚腺苷二磷酸 核糖聚合酶家族(poly(ADP-ribose)polymerases,PARPs)催化完成。PARP1作为PARPs家族最重要的成员,其在DNA损伤应答方面发挥重要作用。研究显示,PARP1在DSBs修复过程中发挥关键作用,参与DSBs的早期应答反应及其具体修复途径,可依据KU蛋白的存在与否发挥不同的特定作用。本文较全面地综述了PARP1在DNA双链断裂修复方面的潜在作用,将为临床疾病的诊治提供新的思路。     

聚腺苷二磷酸核糖基聚合酶──在DNA损伤修复及程序性死亡中的作用

聚腺苷二磷酸核糖基聚合酶(poly (ADP-ribose) polyerase, PARP)是存在于多数真核细胞中的一个蛋白质翻译后修饰酶,它可催化组蛋白H₁等重要核蛋白及它自身的聚腺苷二磷酸核糖基化作用.细胞受到外界损伤因子作用时, DNA发生链断裂,PARP结合到DNA断裂口,其催化活性被激活,修饰受体蛋白,进而引发一系列级联反应.这种性质使PARP有可能作为细胞内的分子感受器和传感器,启动细胞内对损伤作出反应的信号传导机制,从而根据细胞受损程度决定细胞的命运：修复或是死亡．     

聚腺苷二磷酸核糖聚合酶抑制剂耐药性研究进展

聚腺苷二磷酸核糖聚合酶（PARP）抑制剂可选择性杀死同源重组功能缺陷的肿瘤细胞,而对正常细胞的危害较小,这是“合成致死”理论应用于临床的典型范例。尽管 PARP 抑制剂作为一种新型靶向药物,极具应用潜力,但其临床应用也面临诸多问题,其中耐药性的产生被认为是限制 PARP 抑制剂临床应用的重要原因之一。简介 PARP-1 的功能及 PARP-1 抑制剂研究进展,着重综述 PARP-1 抑制剂耐药的临床表现、可能的发生机制及逆转策略,为 PARP-1 抑制剂的临床合理应用提供参考。     

环化二磷酸腺苷核糖及其钙动员的功能

环化二磷酸腺苷核糖（cyclic ADP-ribose,cADPR）是烟酰胺腺嘌呤二核苷酸(NAD⁺)的代谢产物,是新近发现的一种细胞内第二信使．在许多哺乳类和无脊椎动物细胞中,cADPR能引起胞内钙库释放钙离子,其可能机制是：cADPR受体结合cADPR,通过Ryanodine受体或类Ryanodine受体介导的钙通道使cADPR敏感的钙库释放钙离子,此外,一条由一氧化氮（NO）、环化鸟苷酸（cGMP）和cADPR组成的细胞内信号转导途径可能存在于许多细胞中．     

基于小分子化合物库筛选协同多腺苷二磷酸核糖聚合酶抑制剂杀伤卵巢癌细胞的药物

多腺苷二磷酸核糖聚合酶(poly(ADP-ribose) polymerase, PARP)抑制剂是一类靶向 DNA 修复缺陷癌细胞的新型药物。早期研究表明 PARP 抑制剂取得了令人满意的结果,然而药物治疗后出现的耐药机制尚未完全揭露。因此,有必要寻找更多的靶向药物与PARP 抑制剂联用,以达到杀伤肿瘤细胞的目的。本文基于379种小分子化合物和PARP抑制剂尼拉帕尼(Niraparib)的联合用药筛选,通过细胞增殖实验、克隆存活实验和免疫荧光染色等方法筛选潜在的具有协同PARP抑制剂杀伤卵巢癌细胞的药物。结果表明,其中有8种小分子化合物具有较好的联合用药效果,包括2种已经报道的与PARP抑制剂具有联用效果的小分子化合物STF-118804和Disulfiram。我们从中选取原肌球蛋白受体激酶 A (tropomyosin receptor kinase A,TrKA)的抑制剂GW441756,进行了多种肿瘤细胞的验证以及初步机制的探究。Niraparib和TrKA抑制剂的联合用药显著增加肿瘤细胞对PARP抑制剂的敏感性(P<0.05)。从机制上分析,联合用药组细胞内γH2AX foci的数目显著增加(P<0.05),说明TrKA抑制剂阻碍损伤后细胞的DNA损伤修复能力;同时,联合用药显著降低细胞内同源重组修复(homologous recombination repair,HRR)标志物RAD51 foci(P<0.05)的形成,说明TrKA抑制剂可能通过抑制细胞的HRR效率阻碍细胞的DNA损伤修复。本研究的结果提示,TrKA抑制剂可以作为一种与PARP抑制剂联用杀伤卵巢癌细胞的潜在药物。     

Growth-phase-dependent response to DNA damage in poly(ADP-ribose) polymerase deficient cell lines: basis for a new hypothesis describing the role of poly(ADP-ribose) polymerase in DNA replication and repair

We have studied the role of poly(ADP-ribose) polymerase in the repair of DNA damage induced by x-ray and N-methyl N-nitro-N-nitrosoguanidine (MNNG) by using V79 chinese hamster cells, and two derivative mutant cell lines, ADPRT54 and ADPRT351, that are deficient in poly(ADP-ribose) polymerase activity. Under exponentially growing conditions these mutant cell lines are hypersensitive to x-irradiation and MNNG compared to their parental V79 cells which could be interpreted to suggest that poly(ADP-ribose) polymerase is involved in the repair of DNA damage. However, the level of DNA strand breaks induced by x-irradiation and MNNG and their rates of repair are similar in all the cell lines, thus suggesting that it may not be the difference in strand break formation or in its rate of repair that is contributing to the enhanced cell killing in exponentially growing poly(ADP-ribose) polymerase deficient cell lines. In contrast, under growth-arrested conditions, all three cell lines become similarly sensitive to both x-irradiation and MNNG, thus suggesting that poly(ADP-ribose) polymerase may not be involved in the repair of DNA damage in growth-arrested cells. These paradoxical results could be interpreted to suggest that poly(ADP-ribose) polymerase is involved in DNA repair in a cell-cycle-dependent fashion, however, it is functionally active throughout the cell cycle. To resolve this dilemma and explain these results and those obtained by many others, we propose that the normal function of poly(ADP-ribose) polymerase is to prevent DNA recombination processes and facilitate DNA ligation.     

Poly(ADP-ribose) Glycohydrolase Is Present and Active in Mammalian Cells as a 110-kDa Protein

Poly(ADP-ribose) glycohydrolase (PARG) is the major enzyme responsible for the catabolism of poly(ADP-ribose), a reversible covalent-modifier of chromosomal proteins. Purification of PARG from many tissues revealed heterogeneity in activity and structure of this enzyme. To investigate PARG structure and localization, we developed a highly sensitive one-dimensional zymogram allowing us to analyze PARG activity in crude extracts of Cos-7, Jurkat, HL-60, and Molt-3 cells. In all extracts, a single PARG activity band corresponding to a protein of about 110 kDa was detected. This 110-kDa PARG activity was found mainly in cytoplasmic rather than in nuclear extracts of Cos-7 cells.     

Inhibition of poly(ADP-ribosyl)ation in cancer: Old and new paradigms revisited

Inhibitors of poly(ADP-ribose) polymerases actualized the biological concept of synthetic lethality in the clinical practice, yielding a paradigmatic example of translational medicine. The profound sensitivity of tumors with germline BRCA mutations to PARP1/2 blockade owes to inherent defects of the BRCA-dependent homologous recombination machinery, which are unleashed by interruption of PARP DNA repair activity and lead to DNA damage overload and cell death. Conversely, aspirant BRCA-like tumors harboring somatic DNA repair dysfunctions (a vast entity of genetic and epigenetic defects known as “BRCAness”) not always align with the familial counterpart and appear not to be equally sensitive to PARP inhibition. The acquisition of secondary resistance in initially responsive patients and the lack of standardized biomarkers to identify “BRCAness” pose serious threats to the clinical advance of PARP inhibitors; a feeling is also emerging that a BRCA-centered perspective might have missed the influence of additional, not negligible and DNA repair-independent PARP contributions onto therapy outcome. While regulatory approval for PARP1/2 inhibitors is still pending, novel therapeutic opportunities are sprouting from different branches of the PARP family, although they remain immature for clinical extrapolation. This review is an endeavor to provide a comprehensive appraisal of the multifaceted biology of PARPs and their evolving impact on cancer therapeutics.     

Cytoplasmic poly(ADP-ribose) polymerase and poly(ADP-ribose) glycohydrolase in AEV-transformed chicken erythroblasts

Poly(ADP-ribose) polymerase and poly(ADP-ribose) glycohydrolase activities were both investigated in chicken erythroblasts transformed by Avian Erythroblastosis Virus. Respectively 21% and 58% of these activities were found to be present in the post-mitochondrial supernatant (PMS). Fractionation of the PMS on sucrose gradients and poly(A⁺) mRNA detection by hybridization to [³H] poly(U) show that cytoplasmic poly(ADP-ribose) polymerase is exclusively localized in free mRNP. The glycohydrolase activity sedimented mostly in the 6 S region but 1/3 of the activity was in the free mRNP zone. Seven poly(ADP-ribose) protein acceptors were identified in the PMS in the Mr 21000–120000 range. The Mr 120000 protein corresponds to automodified poly(ADP-ribose) polymerase. A Mr 21000 protein acceptor is abundant in PMS and a Mr 34000 is exclusively associated with ribosomes and ribosomal subunits. The existence of both poly(ADP-ribose) polymerase and glycohydrolase activities in free mRNP argues in favour of a role of poly(ADP-ribosylation) in mRNP metabolism. A possible involvement of this post translational modification in the mechanisms of repression-derepression of mRNA is discussed.Abbreviations ADP-ribose adenosine (5) diphospho(5)--D ribose - poly(ADP-ribose) polymer of ADP-ribose - mRNP messenger ribonucleoprotein particles - PMSF phenylmethylsulfonyl fluoride - LDS lithium dodecyl sulfate - TCA trichloroacetic acid