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KREH1 RNA helicase activity promotes utilization of initiator gRNAs across multiple mRNAs in trypanosome RNA editing

Nucleic Acids Research. 2023-05; 
Ashutosh P Dubey, Brianna L Tylec, Amartya Mishra, Katherine Sortino, Runpu Chen, Yijun Sun, Laurie K Read
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Gene Synthesis The recoded KREH1 DNA sequence was synthesized by GenScript containing an in-frame C-terminal 6His-TEV-2myc (HTM) tag Get A Quote


Mitochondrial U-indel RNA editing in kinetoplastid protozoa is directed by trans-acting gRNAs and mediated by a holoenzyme with associated factors. Here, we examine the function of the holoenzyme-associated KREH1 RNA helicase in U-indel editing. We show that KREH1 knockout (KO) impairs editing of a small subset of mRNAs. Overexpression of helicase-dead mutants results in expanded impairment of editing across multiple transcripts, suggesting the existence of enzymes that can compensate for KREH1 in KO cells. In depth analysis of editing defects using quantitative RT-PCR and high-throughput sequencing reveals compromised editing initiation and progression in both KREH1-KO and mutant-expressing cells. In addition,... More