Understanding the Evolution, Biology, and Molecular Mechanism of Argonaute Proteins

了解 Argonaute 蛋白质的进化、生物学和分子机制

• 批准号: 10634674
• 负责人: PHILLIP D ZAMORE
• 金额: $ 50.98万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-03 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634674
• 关键词: Animals; Bacteria; Bacterial Genome; Biochemical; Biochemistry; Biology; Chromosomes; Compensation; Complex; DNA; DNA Sequence; DNA Topoisomerase IV; DNA biosynthesis; Development; Dicer Enzyme; Ensure; Evolution; Exclusion; Gene Expression; Genes; Genetic; Genetic Transcription; Genomic Segment; Germ Cells; Goals; Guide RNA; Male Infertility; Mediating; MicroRNAs; Molecular; Moths; Mus; Mutation; Organism; Pathway interactions; Pharmaceutical Preparations; Plants; Process; Protein Family; Proteins; RNA; RNA Precursors; RNA Sequences; Reproductive Health; Research; Ribosomes; Role; Small RNA; Specificity; Spermatocytes; Thermus thermophilus; Transcript; Translations; Work; computer studies; design; experimental study; fly; fungus; human disease; improved; in vivo; insight; mRNA Stability; mouse genetics; piRNA; placental mammal; pre-miRNA; sperm cell

Project Summary Argonautes are the only known family of proteins that can be programmed with any RNA or DNA sequence to make sequence-specific regulators of transcription, mRNA stability, or translation. Our goal is to understand the biology and mechanism of paradigmatic examples of Argonaute proteins and pathways, and, ultimately, to use these insights to design and improve small RNA-guided therapies for human diseases. Indeed, studying how Argonautes work and how their small RNA guides are made has led to the development and FDA approval of small RNA drugs. Nevertheless, fundamental questions about the specificity and function of Argonaute protein-mediated pathways remain unanswered. Despite >20 years of study, for example, we still cannot predict how Dicer enzymes will cleave a pre- miRNA based only on its sequence. We will use biochemical and structural approaches to identify the features that determine where Dicer cleaves a pre-miRNA and how Dicer partner proteins alter this process. In animals, the PIWI subfamily of Argonaute proteins uses 23–30-nt “piRNA” guides to silence transposons or regulate gene expression in germ cells. piRNAs are made from specific long, single-stranded precursor RNAs. Our research seeks to explain why some genomic regions and transcripts are destined to make piRNAs, while others are excluded. By studying piRNAs in flies, moths, and mice, we hope to identify both evolutionarily ancient and newly evolved strategies that animals use to designate piRNA precursors and to convert them into functional complexes with PIWI proteins. While experimental and computational studies have dramatically improved our ability to predict miRNA targets, similar advances have not yet been made for piRNAs. In the spermatocytes of placental mammals, pachytene piRNAs are nearly as abundant as ribosomes, but we still do not know what or how they regulate. Mutations in the proteins that make pachytene piRNAs cause male infertility, suggesting that pachytene piRNAs promote sperm development. We will use biochemistry and mouse genetics to study the function and specificity of pachytene piRNAs. Finally, 30% of bacterial genomes encode Argonautes, yet we do not know what they do. Surprisingly, we find that in Thermus thermophilus, the DNA-guided, DNA-cleaving Argonaute (TtAgo) acts together with gyrase A to ensure successful replication. Our hypothesis is that TtAgo has acquired a role in disentangling the circular chromosomes at the end of DNA replication, perhaps to compensate for the absence of Topoisomerase IV in this organism. We will use genetics and biochemistry to understand how TtAgo acquires its guides, and how and what it regulates in vivo. Together these studies will reveal diverse strategies that organisms use to make small RNAs and how they use Argonautes to control development, differentiation, and reproductive health.

项目总结

项目成果

Thermus thermophilus Argonaute Functions in the Completion of DNA Replication.

• DOI: 10.1016/j.cell.2020.07.036
• 发表时间: 2020-09-17
• 期刊: Cell
• 影响因子: 64.5
• 作者: Jolly SM;Gainetdinov I;Jouravleva K;Zhang H;Strittmatter L;Bailey SM;Hendricks GM;Dhabaria A;Ueberheide B;Zamore PD
• 通讯作者: Zamore PD

Tetrazine-Ligated CRISPR sgRNAs for Efficient Genome Editing.

• DOI: 10.1021/acschembio.2c00116
• 发表时间: 2022-05-20
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Chen, Zexiang;Devi, Gitali;Arif, Amena;Zamore, Phillip D.;Sontheimer, Erik J.;Watts, Jonathan K.
• 通讯作者: Watts, Jonathan K.

A maternally programmed intergenerational mechanism enables male offspring to make piRNAs from Y-linked precursor RNAs in Drosophila.

• DOI: 10.1038/s41556-023-01227-4
• 发表时间: 2023-10
• 期刊: Nature cell biology
• 影响因子: 21.3

The transcription factor TCFL5 responds to A-MYB to elaborate the male meiotic program in mice.

• DOI: 10.1530/rep-22-0355
• 发表时间: 2023-02-01
• 期刊: Reproduction (Cambridge, England)
• 作者: Cecchini K;Biasini A;Yu T;Säflund M;Mou H;Arif A;Eghbali A;Colpan C;Gainetdinov I;de Rooij DG;Weng Z;Zamore PD;Özata DM
• 通讯作者: Özata DM

A-MYB/TCFL5 regulatory architecture ensures the production of pachytene piRNAs in placental mammals.

• DOI: 10.1261/rna.079472.122
• 发表时间: 2022-10-14
• 期刊: RNA
• 影响因子: 4.5
• 作者: Yu, Tiangxiong;Biasini, Adriano;Cecchini, Katharine;Saflund, Martin;Mou, Haiwei;Arif, Amena;Eghbali, Atiyeh;De Rooij, Dirk G.;Weng, Zhiping;Zamore, Phillip D.;Ozata, Deniz M.
• 通讯作者: Ozata, Deniz M.