1. Academic Validation
  2. Identification of purine biosynthesis as an NADH-sensing pathway to mediate energy stress

Identification of purine biosynthesis as an NADH-sensing pathway to mediate energy stress

  • Nat Commun. 2022 Nov 17;13(1):7031. doi: 10.1038/s41467-022-34850-0.
Ronghui Yang # 1 2 Chuanzhen Yang # 1 2 Lingdi Ma 1 Yiliang Zhao 1 Zihao Guo 1 Jing Niu 1 Qiaoyun Chu 1 Yingmin Ma 2 Binghui Li 3 4
Affiliations

Affiliations

  • 1 Department of Biochemistry and Molecular Biology, Capital Medical University, Beijing, China.
  • 2 Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, China.
  • 3 Department of Biochemistry and Molecular Biology, Capital Medical University, Beijing, China. bli@ccmu.edu.cn.
  • 4 Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, China. bli@ccmu.edu.cn.
  • # Contributed equally.
Abstract

An enhanced NADH/NAD+ ratio, termed reductive stress, is associated with many diseases. However, whether a downstream sensing pathway exists to mediate pathogenic outcomes remains unclear. Here, we generate a soluble pyridine nucleotide transhydrogenase from Escherichia coli (EcSTH), which can elevate the NADH/NAD+ ratio and meantime reduce the NADPH/NADP+ ratio. Additionally, we fuse EcSTH with previously described LbNOX (a water-forming NADH oxidase from Lactobacillus brevis) to resume the NADH/NAD+ ratio. With these tools and by using genome-wide CRISPR/Cas9 library screens and metabolic profiling in mammalian cells, we find that accumulated NADH deregulates PRPS2 (Ribose-phosphate pyrophosphokinase 2)-mediated downstream purine biosynthesis to provoke massive energy consumption, and therefore, the induction of energy stress. Blocking purine biosynthesis prevents NADH accumulation-associated cell death in vitro and tissue injury in vivo. These results underscore the pathophysiological role of deregulated purine biosynthesis in NADH accumulation-associated disorders and demonstrate the utility of EcSTH in manipulating NADH/NAD+ and NADPH/NADP+.

Figures
Products