Project description:Folate metabolism is intricately linked to purine de novo synthesis through the incorporation of folate-derived one-carbon units into the purine scaffold. Here, we investigate the chemical and genetic dependencies caused by mutations in the folate enzyme MTHFD1 and discover a key role for Nudix hydrolase 5 (NUDT5) in regulating purine de novo synthesis. Through genetic knockout and development of a selective chemical NUDT5 degrader, we uncover an unprecedented scaffolding role rather than NUDT5 enzymatic activity is responsible for this phenotype. We find that NUDT5 interacts with the rate-limiting enzyme of purine de novo synthesis, PPAT, to repress the pathway in response to elevated purine levels. Our findings establish NUDT5 as an important regulator of purine de novo synthesis and elucidate its role in mediating sensitivities to purine analogs in cancer treatment and to adenosine in MTHFD1 deficiency.
Project description:Identification of the interaction partners of the protein ecdysoneless (Ecd) in Drosophila melanogaster S2 cells as well as profiling of the changes in binding for mutant, truncated Ecd del34 protein.
Project description:Although ADSL is an enzyme that is involved in de novo purine synthesis, it was reported that ADSL knockdown did not result in decreases in adenine nucleotides (AMP, ADP, and ATP) or GTP. Then, we hypothesized that decreased cell proliferation, migration and invasion capability by ADSL knockdown in endometrial cancer cells were caused through a mechanism independent of purine synthesis. To elucidate a mechanism, we performed DNA microarray-based gene expression profiling using ADSL knockdown and control HEC1B cells. In the analysis, 940 out of 34,127 probes showed greater than two-fold changes in expression with a significant difference, p value < 0.05.
Project description:Reactivating cell cycle re-entry in adult cardiomyocytes improves cardiac function after myocardial infarction (MI). De novo purine synthesis is a critical source of nucleotides and is involved in modulating cell proliferation. Here, using gain-of-function genetic tools, we explored the role of the de novo purine synthesis enzyme Adssl1 in cardiac regeneration. RNA sequencing analysis suggested that Adssl1 overexpression induced strong dedifferentiation and cell cycle entry.
