Generation and application of second messenger molecules by SMODS and SAVED

SMODS和SAVED第二信使分子的生成和应用

基本信息

批准号：
10078261
负责人：
Raven H Huang
金额：
$ 18.87万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10078261
关键词：
Abbreviations Affinity Animals Bacteria Bacterial Infections Bacteriology Biochemical Biochemistry Bioinformatics Biological Biological Process Biology Cell physiology Chemical Structure Complex Cyclic GMP DNA Dinucleoside Phosphates Double-Stranded RNA Enzymes Family Foundations Future Generations Genome Immune response In Vitro Innate Immune Response Investigation Ligase Link Measures Mobile Genetic Elements Monitor Names Neighborhoods Nucleotides Oligonucleotides Operon Organism Pathway interactions Periodicity Phospholipids Play Production Pyrimidine RNA Recombinants Reporting Research Role Second Messenger Systems Specificity Structure System Testing Virus Diseases X-Ray Crystallography base cell growth comparative genomics ds-DNA inorganic phosphate novel therapeutics nucleotidyltransferase oligoadenylate pathogenic bacteria receptor reconstitution sensor structural biology success

项目摘要

Project Summary/Abstract Nucleotide-based second messengers play important biological functions in living organisms. Among various second messenger molecules, several of them are generated by families of NTases that belong to Polb superfamily. In animals, cytosolic NTases OAS and cGAS generate 2’-5’-linked oligoadenylates and 2’- 5’-linked c-GAMP, respectively, both of which trigger innate immune response against viral and bacterial infection. In bacteria, NTase DncV generates 3’-5’-linked cGAMP, which activates CapV to inhibit cell growth. Using a combination of comparative genomics, sequence conservation, and structure analysis, Aravind and coworkers uncovered a vast network of nucleotide-centric systems in bacteria. A family of NTases named SMODS (secondary messenger oligo and dinucleotides synthase) was predicted to generate second messengers. And several conserved domains were predicted to be receptors of the second messengers generated by SMODS. Among the predicted receptors, the domain named SAVED (SMODS-associated and fused to various effector domains) is the most abundant. With the exception of a recent characterization of the product generated by one of SMODS, however, the predicted biochemical and biological functions of SMODS and SAVED have not been experimentally tested. Employing approaches of bioinformatics, biochemistry, and structural biology, we aim to pursue the following two lines of investigation of SMODS and SAVED: 1) We will in vitro reconstitute the enzymatic activity of SMODS and probe interaction between SAVED and the second messengers generated by SMODS; and 2) We will carry out structural studies of SMODS and SAVED, SMODS in complex with its activator and substrates, and SAVED in complex with the second messengers generated by SMODS.

项目概要/摘要基于核苷酸的第二信使在生物体中发挥重要的生物学功能。各种第二信使分子，其中一些是由属于 NTase 家族产生的 Polb 超家族在动物中，胞质 NTase OAS 和 cGAS 产生 2'-5'- 连接的寡腺苷酸和 2'- 分别为 5’-连接的 c-GAMP，两者都会触发针对病毒和细菌的先天免疫反应在细菌感染中，NTase DncV 会生成 3'-5' 连接的 cGAMP，从而激活 CapV 来抑制细胞。结合比较基因组学、序列保守和结构分析，阿拉文德和同事发现了细菌家族中一个庞大的以核苷酸为中心的系统网络。名为 SMODS（第二信使寡核苷酸和二核苷酸合酶）的 NTase 预计会产生第二信使并且几个保守结构域被预测为受体。 SMODS 产生的第二信使在预测的受体中，名为 SAVED 的结构域。（SMODS 相关并与各种效应域融合）是最丰富的，但 a 除外。然而，SMODS 之一生成的产物的最新表征，预测的生化 SMODS和SAVED的生物学功能尚未经过实验测试。生物信息学、生物化学和结构生物学的方法，我们的目标是追求以下两条路线 SMODS 和 SAVED 的研究： 1) 我们将在体外重建 SMODS 的酶活性，探测 SAVED 和 SMODS 生成的第二信使之间的相互作用，并且 2) 我们将进行；进行 SMODS 和 SAVED 的结构研究，SMODS 与其激活剂和底物的复合物，以及与 SMODS 生成的第二信使一起保存在复合体中。