tREPs-A New Class of Functional tRNA-Encoded Peptides

Area of Science:

• Biochemistry
• Molecular Biology
• Parasitology

Background:

• Transfer RNA (tRNA) sequences are primarily known for their role in protein synthesis.
• The potential for tRNA sequences to encode functional peptides (tREPs) has been largely unexplored.

Purpose of the Study:

• To investigate the potential for tRNA sequences to encode functional peptides.
• To artificially synthesize and characterize novel tREPs for therapeutic applications.
• To identify and validate tREPs with antiparasitic activity.

Main Methods:

• Computational translation of Escherichia coli tRNA sequences into hypothetical tREP sequences.
• Bioinformatic filtering (sequence, structure, energy) to select candidate tREPs.
• Chemical synthesis of selected tREPs, including tREP-18.
• In vitro antileishmanial assays against Leishmania donovani strains (Ag83 and BS12).
• Microscopy (AFM, SEM) to assess structural changes in parasites.
• Mitochondrial membrane potential assays.

Main Results:

• A database of hypothetical tREPs was generated from E. coli tRNA sequences.
• The synthesized tREP-18 demonstrated potent antileishmanial activity against Leishmania donovani (IC50 = 22.13 nM for Ag83, 15 nM for BS12).
• tREP-18 induced significant alterations in parasite cytoskeletal architecture and destabilized mitochondrial membrane potential.
• tREP-18 showed efficacy against the amastigote stage and promoted macrophage pathogen clearance.

Conclusions:

• This study provides proof-of-concept for generating a new class of functional peptides from tRNA sequences.
• tREP-18 represents a novel, potent antileishmanial agent with a unique mechanism of action.
• The exploration of tRNA-peptide space offers a vast, untapped resource for discovering novel therapeutic agents.