Riboregulation of Enolase 1 activity controls glycolysis and embryonic stem cell differentiation

Area of Science:

• Biochemistry
• Molecular Biology
• Developmental Biology

Background:

• Stem cell differentiation necessitates precise coordination between metabolic pathways and cell fate determination.
• Glycolysis, a central metabolic pathway, plays a critical role in regulating stem cell functions.

Purpose of the Study:

• To investigate the regulatory mechanisms linking metabolism and cell fate in differentiating stem cells.
• To identify novel regulators of glycolytic enzymes involved in stem cell differentiation.

Main Methods:

• In vitro enzymatic assays to assess Enolase 1 (ENO1) activity.
• RNA-protein interaction studies to identify RNA ligands binding to ENO1.
• Cell culture experiments with human cells and mouse embryonic stem cells (mESCs).
• Pharmacological and RNAi-mediated modulation of SIRT2 and ENO1.
• Analysis of ENO1 acetylation and its impact on RNA binding and enzymatic activity during mESC differentiation.

Main Results:

• Specific RNA ligands were identified that inhibit ENO1 activity in vitro and reduce glycolysis in cells.
• SIRT2 activity inversely correlates with ENO1 acetylation and RNA binding.
• mESC differentiation induces ENO1 acetylation, enhancing RNA binding and inhibiting glycolysis.
• Mutant ENO1 forms that bypass this regulation impair germ layer differentiation.

Conclusions:

• Acetylation-driven riboregulation of ENO1 is a key mechanism controlling glycolysis and stem cell differentiation.
• This RNA-based regulation of enzyme activity (riboregulation) may be a widespread biological control principle.
• Findings provide insights into metabolic control during stem cell fate decisions.