Targeting the conserved active site of splicing machines with specific and selective small molecule modulators

Area of Science:

• Molecular Biology
• RNA Biology
• Biochemistry

Background:

• Group II introns are ancient RNA enzymes vital in bacteria and organelles.
• They are precursors to the spliceosome and important biotechnological tools.
• Understanding their mechanism is key for therapeutic development.

Purpose of the Study:

• To investigate how group II introns recognize and bind small molecules.
• To elucidate the mechanism by which small molecules inhibit RNA splicing.
• To provide a foundation for designing novel splicing modulators.

Main Methods:

• Enzymatic assays to measure splicing activity.
• Crystallography to determine structural interactions.
• Computational simulations to model molecular dynamics.

Main Results:

• Group II introns recognize small molecules via their conserved active site.
• RNA-binding small molecules inhibit splicing by adopting specific poses.
• Inhibitors prevent essential conformational changes required for catalysis.

Conclusions:

• The conserved RNA core of splicing machinery can specifically bind small molecules.
• This finding supports the rational design of inhibitors for bacterial, organellar, and human spliceosomes.
• Splicing modulators offer potential therapeutic strategies for diseases and cancers.