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Guide-substrate base-pairing requirement for box H/ACA RNA-guided RNA pseudouridylation
Meemanage D De Zoysa1, Guowei Wu1, Raviv Katz1
1Department of Biochemistry and Biophysics, Center for RNA Biology, University of Rochester Medical Center, Rochester, New York 14642, USA.
RNA (New York, N.Y.)
|June 7, 2018
Summary
Box H/ACA RNAs guide RNA pseudouridylation, a crucial process. This study identifies the minimum 8 base pairs needed for specificity and finds the pseudouridylation pocket accommodates sequence variations.
Area of Science:
- Molecular Biology
- RNA Biology
- Biochemistry
Background:
- Box H/ACA RNAs are small RNAs essential for guiding RNA pseudouridylation in eukaryotes and archaea.
- They possess a conserved hairpin-hinge-hairpin-tail structure and primarily direct modifications of rRNA and snRNA.
- The precise base-pairing rules governing guide RNA-substrate interactions for pseudouridylation specificity remain incompletely understood.
Purpose of the Study:
- To investigate the base-pairing interactions between box H/ACA RNAs and their RNA substrates.
- To determine the minimum number of base pairs required for efficient and specific RNA pseudouridylation.
- To assess the flexibility of the box H/ACA RNA pseudouridylation pocket in accommodating diverse substrate sequences.
Main Methods:
- Detailed investigation of guide-substrate base-pairing interactions.
- Analysis of pseudouridylation activity across different box H/ACA RNA pockets.
- Experimental validation of base-pairing requirements and pocket flexibility.
Main Results:
- Identified a minimum of 8 base pairs as essential for RNA-guided pseudouridylation.
- Demonstrated that the pseudouridylation pocket exhibits flexibility, tolerating slight variations in substrate RNA sequences.
- Findings were consistent across three distinct pseudouridylation pockets, indicating general applicability.
Conclusions:
- Established the minimal base-pairing requirement for box H/ACA RNA-guided pseudouridylation.
- Characterized the sequence flexibility of the pseudouridylation pocket, crucial for substrate recognition.
- Provided generalizable insights into the mechanism of box H/ACA RNA-guided RNA modification.
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