NOVEL TRANSPOSONS AND PLASTICITY ZONES OF HELICOBACTER PYLORI

幽门螺杆菌的新型转座子和可塑性区

• 批准号: 8037715
• 负责人: DOUGLAS Eugene BERG
• 金额: $ 18.81万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-05 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8037715
• 关键词: Affect; Bacterial Chromosomes; Bacterial DNA; Bacterial Drug Resistance; Binding; Biological Assay; Cell Culture Techniques; Cell division; Cells; Chronic; Comparative Study; DNA; DNA Methylation; DNA Modification Process; DNA Transposable Elements; Data; Development; Diagnosis; Dimensions; Disease; Embryonic Development; Ensure; Epigenetic Process; Epithelial Cells; Escherichia coli; Evolution; Excision; Family; Gene Expression; Gene Expression Regulation; Genes; Genetic Recombination; Genetic Transcription; Genome; Grant; Health; Helicobacter pylori; High-Risk Cancer; Human; Immune system; Infection; Laboratories; Length; Mammalian Cell; Mediating; Methylation; Methyltransferase; Movement; Nucleic Acids; Oncogenic; Organism; Outcome; Pathology; Phenotype; Process; Proteins; Public Health; RNA Helicase; Regulation; Retroviridae; Role; Site; Specificity; Stomach; Structure; System; Testing; Tissues; Transposase; Type IV Secretion System Pathway; Tyrosine; V(D)J Recombination; Virulence; Work; bacterial resistance; base; design; disorder risk; expression cloning; genome sequencing; human disease; improved; insight; macromolecule; malignant stomach neoplasm; member; microbial; novel; pathogen; programs; promoter; protein complex; prototype; public health relevance; recombinase; stem; tumorigenesis

DESCRIPTION (provided by applicant): Transposable elements (TEs) contribute importantly to genome organization and evolution, regulation, developmental programming, and human health and disease -- most famously, as bacterial drug resistance transposons, oncogenic retroviruses, and V(D)J immune system recombination. TEs are diverse in mechanisms and regulation of movement, and their analyses provide valuable insights into protein-nucleic acid interactions and cellular regulatory mechanisms. TEs in pathogens often contain auxiliary genes that affect phenotypes such as virulence. This proposal stems from our discovery of novel "plasticity zone" transposons (TnPZs) in the gastric pathogen Helicobacter pylori. These TEs encode a novel member of the XerC/XerD branch of the tyrosine recombinase family ("XerT"), a type IV secretion system ("tfs3"), and a large protein (OrfQ) (2800 - 4000 residues, depending on strain) that contains prominent DNA methylase and RNA helicase motifs. OrfQ-like proteins are evident in genome sequences of other unrelated bacterial pathogens. We hypothesize that (i) TnPZs are conjugative transposons, passed between bacterial cells via their Tfs3 protein complex, and inserted into new sites using XerT protein; and (ii) that the putative DNA methylase OrfQ may cause epigenetic (DNA modification) changes in target tissues that could impact on gastric pathology and disease. In Specific Aim 1 we will study the mechanism and control of TnPZ excision and transposition in H. pylori and E. coli. Since prototype (E. coli) XerC/XerD proteins are specific for one unique chromosomal sequence whereas TnPZs insert into many sites, studies of XerT action should enhance understanding of protein-nucleic acid specificity and its evolution. In Specific Aim 2 we will test for OrfQ-mediated DNA methylation in infected mammalian cells and in bacterial cells. OrfQ's domain structure suggests that studies of its action could give new insights into the dynamics of bacterial-host interactions during chronic infection. In conclusion, results of these R21 studies should enhance understanding of transposition-related phenomena, and of infection and virulence mechanisms, and also provide data needed to support an anticipated larger RO1-type application in coming years. PUBLIC HEALTH RELEVANCE: The fundamental insights to be gained from transposon TnPZ studies will enrich understanding of microbial pathogen evolution and human disease. Studies of OrfQ, in particular, could reveal a new dimension of bacterial-host interactions and the origin of human epigenetic changes important in disease pathology, and result in improved diagnosis and therapy for infections and associated pathologies, including gastric cancer.

描述（由申请人提供）： 转座元素（TES）对基因组组织和进化，调节，发育计划以及人类健康和疾病的重要贡献 - 最著名的是细菌耐药性转座子，致癌逆转录病毒以及V（d）J免疫系统重组。 TE在机制和运动的调节方面都是多种多样的，它们的分析为蛋白质核酸相互作用和细胞调节机制提供了宝贵的见解。病原体中的TE通常包含影响毒力等表型的辅助基因。该建议源于我们发现胃病原体幽门螺杆菌中新型的“可塑性区”转座子（TNPZ）。这些TE编码酪氨酸重组酶家族（“ XERT”）的XERC/XERD分支的新成员，IV型分泌系统（“ TFS3”），以及一个大蛋白（ORFQ）（2800-4000个残基，取决于菌株），这些残基，取决于菌株，其含有突出的DNA甲基酶和RNA硫酸盐想要的菌株。在其他无关细菌病原体的基因组序列中，ORFQ样蛋白很​​明显。我们假设（i）TNPZ是共轭转座子，通过其TFS3蛋白复合物在细菌细胞之间传递，并使用XERT蛋白插入新位点； （ii）假定的DNA甲基酶ORFQ可能会导致靶组织的表观遗传学（DNA修饰）变化，这可能会影响胃病理学和疾病。在特定目标1中，我们将研究幽门螺杆菌和大肠杆菌中TNPZ切除和转座的机理和控制。由于原型（大肠杆菌）XERC/XERD蛋白是特异性的，对于一种独特的染色体序列，而TNPZ插入了许多位点，因此对XERT作用的研究应增强对蛋白质核酸特异性及其进化的理解。在特定目标2中，我们将测试感染的哺乳动物细胞和细菌细胞中ORFQ介导的DNA甲基化。 ORFQ的域结构表明，对其作用的研究可以为慢性感染期间细菌 - 宿主相互作用的动态提供新的见解。总之，这些R21研究的结果应增强对转座相关现象以及感染和毒力机制的理解，并提供所需的数据，以支持未来几年预期的较大的RO1型应用。 公共卫生相关性： 从跨盆地TNPZ研究中获得的基本见解将丰富对微生物病原体进化和人类疾病的理解。特别是对ORFQ的研究可能揭示了细菌宿主相互作用的新维度以及人类表观遗传学变化在疾病病理学中重要的重要性，并改善了包括胃癌在内的感染和相关病理学的诊断和治疗。

项目成果

Intact cag pathogenicity island of Helicobacter pylori without disease association in Kolkata, India.

• DOI: 10.1016/j.ijmm.2010.10.003
• 发表时间: 2011-04
• 期刊: INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY
• 影响因子: 4.1
• 作者: Patra, Rajashree;Chattopadhyay, Santanu;De, Ronita;Datta, Simanti;Chowdhury, Abhijit;Ramamurthy, T.;Nair, G. Balakrish;Berg, Douglas E.;Mukhopadhyay, Asish K.
• 通讯作者: Mukhopadhyay, Asish K.

Xer recombinase and genome integrity in Helicobacter pylori, a pathogen without topoisomerase IV.

• DOI: 10.1371/journal.pone.0033310
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Debowski AW;Carnoy C;Verbrugghe P;Nilsson HO;Gauntlett JC;Fulurija A;Camilleri T;Berg DE;Marshall BJ;Benghezal M
• 通讯作者: Benghezal M

Complete Genome Sequences of Two Helicobacter pylori Strains from a Canadian Arctic Aboriginal Community.

来自加拿大北极原住民社区的两种幽门螺杆菌菌株的完整基因组序列。

• DOI: 10.1128/genomea.00209-15
• 发表时间: 2015
• 期刊: Genome announcements
• 作者: Kersulyte,Dangeruta;Bertoli,MTeresita;Tamma,Sravya;Keelan,Monika;Munday,Rachel;Geary,Janis;VeldhuyzenvanZanten,Sander;Goodman,KarenJ;Berg,DouglasE
• 通讯作者: Berg,DouglasE