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Post-transcriptional Gene Regulation by Cytoplasmic Poly(ADP-ribose) Polymerases

细胞质聚（ADP-核糖）聚合酶的转录后基因调控

基本信息

批准号：
9234547
负责人：
Anthony K L Leung
金额：
$ 34.74万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2020-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9234547
关键词：
Ablation Antiviral Agents Antiviral Response Binding Binding Proteins Biochemical Biochemistry Biogenesis Biological Assay Biology Cell Nucleus Cell physiology Cells Clinical Trials Collaborations Cytoplasm Cytoplasmic Granules DNA DNA Modification Process DNA Repair Data Development Ebola virus Embryo Ensure Enzymes Funding Gene Expression Gene Expression Regulation Genetic Transcription HIV Hepatitis B Human In Vitro Individual Integration Host Factors Laboratories Malignant Neoplasms Mass Spectrum Analysis Mediating Messenger RNA Metabolism MicroRNAs Modeling Mus Nuclear Nucleotides Organism Pharmacologic Substance Physiological Play Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Polymerase Polymers Polynucleotides Post-Translational Protein Processing Protein Isoforms Proteins Proteome Proteomics RNA RNA Binding RNA-Binding Proteins Regulator Genes Reporter Role Sindbis Virus Site Stress Surface System Tankyrase Techniques Testing Therapeutic Time Universities Untranslated RNA Viral Viral Physiology Virus Diseases Washington Zinc biological adaptation to stress inhibitor/antagonist innovation novel nucleic acid binding protein overexpression public health relevance response success

项目摘要

DESCRIPTION (provided by applicant): Gene expression is mediated by DNA- and RNA-binding proteins in every organism. Regulation of gene expression is commonly mediated by protein modifications of these nucleic acid binding proteins. This proposal focuses on an under-explored but therapeutically important protein modification called poly(ADP- ribose) (PAR). PAR has been well known for its roles in DNA repair and transcription in the nucleus. Recently, we discovered that PAR also modifies several post-transcriptional mRNA gene regulators in the cytoplasm. Our data are consistent with recent proteomics studies showing that poly(ADP-ribosyl)ated (PARylated) proteomes are enriched with RNA-binding proteins, suggesting that PAR plays a much broader regulatory role in RNA metabolism than previously appreciated. In this proposal, we will focus on how PAR regulates microRNA functions. MicroRNAs are a class of ~22 nucleotide non-coding RNAs that regulate many fundamental cellular processes, including stress responses. Although much has been characterized about microRNA biogenesis, little is known about how microRNA activities are regulated. Key data: Recent data including ours indicate that microRNA activities are inhibited by the PARylation of the core microRNA-binding protein Argonaute (AGO). Such inhibition is regulated by PAR polymerase 13 (PARP- 13) where its overexpression reduces microRNA activities. Intriguingly, PARP-13 is catalytically inactive; therefore, other catalytically active PARP(s) must be involved. Such a PARylation mechanism involving more than one PARP represents a new paradigm. In this proposal, we will investigate how PARP-13 interacts with a cytoplasmic, catalytically active PARP to PARylate AGO (Aim 1), determine how PAR polymers on AGO reduce microRNA activities (Aim 2) and identify which domains of AGO are modified by PAR (Aim 3). We will use a novel mass spectrometry technique to identify AGO PARylation sites. Until now, the identification of PARylation sites has been a challenge for the field and thus this study allows the first systematic analysis of the functional roles of individual sites of a protein substrate. O note, PARP-13 is also known as zinc antiviral protein (ZAP) - a host factor that inhibits the replication of Sindbis virus, Ebola virus, Hepatitis B virus and HIV upon overexpression. Therefore, AGO PARylation-mediated inhibition of microRNA activities may be involved in host antiviral responses, which we will explore in the context of Sindbis virus infection (Aim 3c) in collaboration with Dr. Diane Griffin at Johns Hopkins. The Team: To ensure success, this project is performed with two key collaborators (both of whom we are requesting for one funding module): AGO biochemistry expert Dr. Leemor Joshua-Tor (Cold Spring Harbor Laboratory) and proteomics expert Dr. Shao-En Ong (University of Washington). Other consultants include Drs. Phillip Sharp (MIT) and Carl Novina (Harvard) on microRNA biology, Drs. Ted Dawson (John Hopkins) and Paul Chang (MIT) on PAR biology, and Dr. Pierre Coulombe (Johns Hopkins) on biochemistry.