InsP7 is a small-molecule regulator of NUDT3-mediated mRNA decapping and processing-body dynamics

Soumyadip Sahu, Zhenzhen Wang, Xinfu Jiao, Chunfang Gu, Nikolaus Jork, Christopher Wittwer, Xingyao Li, Sarah Hostachy, Dorothea Fiedler, Huanchen Wang, Henning J. Jessen, Megerditch Kiledjian, Stephen B. Shears

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Regulation of enzymatic 5′ decapping of messenger RNA (mRNA), which normally commits transcripts to their destruction, has the capacity to dynamically reshape the transcriptome. For example, protection from 5′ decapping promotes accumulation of mRNAs into processing (P) bodies-membraneless, biomolecular condensates. Such compartmentalization of mRNAs temporarily removes them from the translatable pool; these repressed transcripts are stabilized and stored until P-body dissolution permits transcript reentry into the cytosol. Here, we describe regulation of mRNA stability and P-body dynamics by the inositol pyrophosphate signaling molecule 5-InsP7 (5-diphosphoinositol pentakisphosphate). First, we demonstrate 5-InsP7 inhibits decapping by recombinant NUDT3 (Nudix [nucleoside diphosphate linked moiety X]-type hydrolase 3) in vitro. Next, in intact HEK293 and HCT116 cells, we monitored the stability of a cadre of NUDT3 mRNA substrates following CRISPRCas9 knockout of PPIP5Ks (diphosphoinositol pentakisphosphate 5-kinases type 1 and 2, i.e., PPIP5K KO), which elevates cellular 5-InsP7 levels by two- to threefold (i.e., within the physiological rheostatic range). The PPIP5K KO cells exhibited elevated levels of NUDT3 mRNA substrates and increased P-body abundance. Pharmacological and genetic attenuation of 5-InsP7 synthesis in the KO background reverted both NUDT3 mRNA substrate levels and P-body counts to those of wild-type cells. Furthermore, liposomal delivery of a metabolically resistant 5-InsP7 analog into wild-type cells elevated levels of NUDT3 mRNA substrates and raised P-body abundance. In the context that cellular 5-InsP7 levels normally fluctuate in response to changes in the bioenergetic environment, regulation of mRNA structure by this inositol pyrophosphate represents an epitranscriptomic control process. The associated impact on P-body dynamics has relevance to regulation of stem cell differentiation, stress responses, and, potentially, amelioration of neurodegenerative diseases and aging.

Original languageEnglish (US)
Pages (from-to)19245-19253
Number of pages9
JournalProceedings of the National Academy of Sciences of the United States of America
Volume117
Issue number32
DOIs
StatePublished - Aug 11 2020

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • Cellular homeostasis
  • Inositol
  • P bodies
  • Signaling

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