Clinically relevant benzoxaboroles inhibit mRNA processing in Trypanosoma brucei

Area of Science:

• Molecular Biology
• Parasitology
• Drug Discovery

Background:

• The CPSF73 endonuclease is a potential target for benzoxaborole anti-trypanosomal drugs.
• Previous studies indicated AN7973 inhibits mRNA processing.
• Acoziborole and AN11736 are drug candidates for human sleeping sickness and nagana in cattle, respectively.

Purpose of the Study:

• To investigate if acoziborole and AN11736 inhibit mRNA processing in trypanosomes.
• To analyze protein interactions with CPSF73 after AN7973 treatment using mass spectrometry.

Main Methods:

• Assessed mRNA processing by measuring spliced mRNA levels and polycistronic transcripts.
• Used affinity purification to isolate tagged CPSF73 from treated and untreated trypanosomes.
• Employed mass spectrometry to identify differentially co-purified proteins.

Main Results:

• AN11736 and acoziborole significantly inhibited mRNA processing.
• Inhibition was evidenced by reduced spliced mRNAs and increased di- and tri-cistronic alpha-beta tubulin mRNAs.
• Short-term (30 min) AN7973 treatment did not significantly alter CPSF73-associated proteins.

Conclusions:

• Acoziborole and AN11736, like AN7973, target mRNA processing in trypanosomes.
• These findings support the role of CPSF73 as a target for benzoxaborole drugs.
• Further investigation is needed to understand the precise mechanism and short-term effects of AN7973 on CPSF73 interactions.