Role of Nicotinamide Nucleotide Adenylyltransferase-1 (NMNAT1) in Chromatin Remodeling and Gene Regulation

烟酰胺核苷酸腺苷酸转移酶-1 (NMNAT1) 在染色质重塑和基因调控中的作用

• 批准号: 10607072
• 负责人: Yessenia Cedeno
• 金额: $ 4.77万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607072
• 关键词: ATAC-seq; Address; Binding; CDKN1A gene; CRISPR/Cas technology; Cell Cycle Arrest; Cell Line; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromatin Fiber; Chromatin Structure; Color; Data; Defect; Disease; Enzymes; Functional disorder; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Genome; Genomic approach; Genomics; Human Pathology; Impairment; Kinetics; Knock-out; Knowledge; Lead; Ligase; Malignant Neoplasms; Mediating; Mentorship; Metabolic; Metabolism; Microscopy; Modeling; Molecular; Molecular and Cellular Biology; Morphology; Mutation; Nicotinamide-Nucleotide Adenylyltransferase; Nuclear; Phenotype; Power Plants; Process; Production; Publishing; Regulation; Research; Role; SIRT1 gene; Scientist; Series; Structure; System; Training; Work; age related; career; chromatin remodeling; enzyme activity; experimental study; genomic locus; human disease; imaging approach; imaging system; insight; multidisciplinary; mutant; novel; particle; single molecule; transcriptome; transcriptome sequencing

PROJECT SUMMARY Chromatin remodeling and gene expression are highly dynamic processes in the nucleus. The regulation of these processes is crucial to maintain proper cellular functions. Chromatin remodeling enzymes mediate changes in chromatin structure and control the cycling of chromatin-associated factors on and off the genome. The remodeling of chromatin depends on metabolites (e.g., ATP and NAD+), which fuels chromatin remodeling enzymes activity. NMNAT1 is a metabolic enzyme primarily involved in the local production of NAD+ in the nucleus. NMNAT1 is thought to provide pools of NAD+ which fuels NAD+-dependent chromatin remodeling enzymes, including SIRT1 and PARP1. However, various studies suggest that NMNAT1 may also allosterically regulate these enzymes activity in an enzymatic independent manner. Nonetheless, the role of NMNAT1 in regulating chromatin remodeling and gene expression through local NAD+ production and interactions with chromatin remodeling enzymes remains poorly defined. In this proposal, I seek to define the role of NMNAT1 in regulating chromatin remodeling enzymes activity, chromatin structure, and gene expression. I hypothesize that the cycling of NMNAT1 on the genome produces local NAD+ gradients that regulate chromatin-modifying enzyme activity, influencing rapid changes in chromatin structure and gene expression of target genes. To address this hypothesis, I will (1) establish an NMNAT1 knock-out cellular system to perform structure-function studies in live cells and identify phenotypical defects upon depletion of NMNAT1. To validate this system, I will perform rescue experiments with wild-type NMNAT1 and catalytic dead mutant NMNAT1. (2) Examine how NMNAT1 regulates the expression of chromatin remodeling enzymes (e.g., SIRT1) target genes by defining changes in chromatin structure, gene expression and genomic occupancy, and (3) establish how NMNAT1 cycling dynamics regulate chromatin remodelers and transcriptional bursting. These studies will be essential to determine how nuclear metabolic enzymes like NMNAT1 and metabolite production regulate chromatin structure and gene expression of target genes. This knowledge will define new paradigms regarding gene expression and regulation. Under the mentorship of Drs. Robert Coleman and Robert H. Singer, Yessenia Cedeno Cedeno will receive multi-disciplinary training in molecular and cellular biology combined with expertise in state-of-the-art microscopy. This will allow Yessenia to successfully execute the proposed research and training plans and advance her professional career as an independent scientist.

项目总结