Chromatin structure and control of gene expression in the human malaria parasite

人类疟疾寄生虫的染色质结构和基因表达控制

基本信息

批准号：
10394336
负责人：
Karine Gaelle Le Roch
金额：
$ 67.04万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-02 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10394336
关键词：
3-Dimensional Address Affect Antimalarials Architecture Biological Process Biology Cell Nucleus Cessation of life Chromatin Chromatin Structure Chromosomes Complex Consensus Country Coupled DNA Data Development Eukaryotic Cell Female Gene Expression Gene Expression Regulation Genes Genetic Transcription Genome Genomics Goals Heterochromatin Hi-C Human Infection Infectious Agent Knowledge Laboratories Life Cycle Stages Maintenance Malaria Mass Spectrum Analysis Mediating Methodology Molecular Molecular Genetics Oligonucleotides Outcomes Research Parasites Pathway interactions Plasmodium Plasmodium falciparum Plasmodium falciparum genome Play Population Process Protein Serine/Threonine Phosphatase Protein phosphatase Proteins RNA RNA purification Research Role Scientist Sex Differentiation Sexual Development Structure Testing Therapeutic Transcriptional Regulation Untranslated RNA Virulence chromosome conformation capture design disease transmission drug development enhancer-binding protein AP-2 genetic approach genetic information genome editing genome-wide insight interest knock-down knockout gene male multidisciplinary next generation sequencing novel pathogen programs protein complex protein function sequencing platform transcription factor transmission process

项目摘要

Abstract: Nearly half of the world's population lives in countries where malaria is endemic. Plasmodium falciparum, the causative agent of the most severe form of human malaria, is responsible for 95% of malaria deaths worldwide. This project's main goals are to characterize the molecular determinants that control chromatin organization and gene regulation in this pathogen; elucidate their importance in parasite development, virulence, and sexual differentiation; and identify novel pathways that can be targeted to kill the parasite. The proposed research builds upon a large body of molecular, cellular, and genome-wide data generated in the Pl's lab that discovered how close interconnections between 3D genome organization and transcription regulate sexual differentiation and parasite development. Despite significant progress in the past 10 years in elucidating the role of chromatin in transcriptional control, the mechanism underlying changes in chromatin structure and the factors controlling these changes remain to be elucidated. The studies proposed here will examine the proteins and long non-coding RNAs (lncRNAs) involved in the control of chromatin structure, parasite development, virulence, and sexual differentiation. The project is organized into three Specific Aims. Aim 1 will validate the role and essentiality of two proteins that we previously identified as potential master regulators of male and female sexual differentiation. Two genes encoding an AP2 transcription factor (PfApi2AP2-FG) and a serine/threonine protein phosphatase 2A activator (PfPTPA) are located at the boundary of two condensed chromatin super domains observed in both early and late gametocytes. We will fully investigate the function of these proteins during male and female gametocyte development using cellular, molecular, and genetic approaches. Aim 2 will validate the role of two lncRNAs (lncRNAG9 and lncRNAG14) that we previously identified as potential regulators of sexual differentiation and chromosome reorganization during parasite development. These factors may be involved in chromosome reorganization in gametocyte stages. We will therefore implement a set of cellular, molecular, and genetic approaches to characterize their role in the formation and maintenance of parasite sexual stages, as well as in the specific chromosomal reorganization we observed at these stages. Aim 3 will systematically isolate and identify the proteins and lncRNAs that control the structure and activity of transcriptionally silent heterochromatin clusters in P. falciparum genome, using a novel methodology called chromatin isolation by RNA purification (ChlRP). Once identified, factors will be validated at the functional level using cellular and molecular experimental approaches, including genome editing by CRlSPR-Cas9. lt is anticipated that the proposed research will offer groundbreaking new insights into parasite-specific protein complexes and lncRNAs and their role in chromatin structure and parasite biology, as well as many starting points for novel directions in malaria research and therapy.

抽象的：世界上将近一半的人口生活在疟疾流行的国家。疟原虫恶性菌是最严重的人类疟疾的病因，是负责全球95％的疟疾死亡。这个项目的主要目标是表征控制染色质组织和基因的分子决定因素在这种病原体中调节；阐明它们在寄生虫发展，毒力，毒力中的重要性和性别差异；并确定可以针对杀死的新颖途径寄生虫。拟议的研究建立在大量分子，细胞和在PL的实验室中生成的全基因组数据，该数据发现3D之间的互连如何接近基因组组织和转录调节性区分和寄生虫发展。尽管在过去的10年中取得了重大进展，但在阐明染色质的作用转录控制，染色质结构变化的机制和因素控制这些变化仍有待阐明。这里提出的研究将检查蛋白质和长期非编码RNA（LNCRNA）涉及染色质的控制结构，寄生虫的发展，毒力和性别分化。该项目是组织成三个特定目标。 AIM 1将验证我们以前确定为男性潜在的主调节剂的两种蛋白质和女性性别差异。编码AP2转录因子的两个基因（PFAPI2AP2-FG）和丝氨酸/苏氨酸蛋白磷酸酶2a激活剂（PFPTPA）为位于两个在两个中观察到的两个冷凝染色质超级域的边界早期和晚配子细胞。我们将充分研究这些蛋白质的功能在雄性和女性配子细胞发育期间，使用细胞，分子和遗传方法。 AIM 2将验证两个LNCRNA（lncrnag9和lncrnag14）的作用先前被确定为性别分化和染色体的潜在调节剂寄生虫开发过程中的重组。这些因素可能涉及染色体在配子细胞阶段进行重组。因此，我们将实施一组蜂窝分子和遗传方法来表征它们在形成和维持中的作用我们在这些阶段。 AIM 3将系统地隔离并识别蛋白质和lncRNA 控制转录静音异染色质的结构和活动恶性疟原虫基因组中的簇，使用一种称为染色质分离的新方法通过RNA纯化（CHLRP）。一旦确定，将在使用细胞和分子实验方法（包括基因组）的功能水平 CRLSPR-CAS9编辑。预计拟议的研究将提供对寄生虫特异性蛋白复合物和lncrnas及其的开创性新见解及其在染色质结构和寄生虫生物学中的作用，以及许多新颖的起点疟疾研究和治疗方向。

项目成果

期刊论文数量（33）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Attacking COVID-19 Progression Using Multi-Drug Therapy for Synergetic Target Engagement.

DOI：
10.3390/biom11060787
发表时间：
2021-05-23
期刊：
Biomolecules
影响因子：
5.5
作者：
Coban MA;Morrison J;Maharjan S;Hernandez Medina DH;Li W;Zhang YS;Freeman WD;Radisky ES;Le Roch KG;Weisend CM;Ebihara H;Caulfield TR
通讯作者：
Caulfield TR

Strand-Specific RNA-Seq Applied to Malaria Samples.

链特异性 RNA 测序应用于疟疾样本。

DOI：
10.1007/978-1-0716-0743-5_2
发表时间：
2021
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Lu,XueqingMaggie;LeRoch,Karine
通讯作者：
LeRoch,Karine

Dynamic Chromatin Structure and Epigenetics Control the Fate of Malaria Parasites.

DOI：
10.1016/j.tig.2020.09.003
发表时间：
2021-01
期刊：
Trends in genetics : TIG
影响因子：
0
作者：
Hollin T;Gupta M;Lenz T;Le Roch KG
通讯作者：
Le Roch KG

Three-dimensional chromatin in infectious disease-A role for gene regulation and pathogenicity?

DOI：
10.1371/journal.ppat.1009207
发表时间：
2021-03
期刊：
PLoS pathogens
影响因子：
6.7
作者：
Davis SZ;Hollin T;Lenz T;Le Roch KG
通讯作者：
Le Roch KG

The Arabidopsis RRM domain protein EDM3 mediates race-specific disease resistance by controlling H3K9me2-dependent alternative polyadenylation of RPP7 immune receptor transcripts.

DOI：
10.1111/tpj.14148
发表时间：
2019-03
期刊：
The Plant journal : for cell and molecular biology
影响因子：
0
作者：
Lai Y;Cuzick A;Lu XM;Wang J;Katiyar N;Tsuchiya T;Le Roch K;McDowell JM;Holub E;Eulgem T
通讯作者：
Eulgem T