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RNA Pull-down Procedure to Identify RNA Targets of a Long Non-coding RNA
Published on: April 10, 2018
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Functional and molecular dissection of HCMV long non-coding RNAs
Sungwon Lee1,2, Hyewon Kim1,2, Ari Hong2,3
1School of Biological Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
Scientific Reports
|November 12, 2022
Summary
Human cytomegalovirus (HCMV) expresses essential long non-coding RNAs (lncRNAs) that regulate viral replication and latency. N6-methyladenosine (m6A) modification stabilizes these viral lncRNAs, explaining their high abundance.
Area of Science:
- Virology
- Molecular Biology
- RNA Biology
Background:
- Human cytomegalovirus (HCMV) defies genome economy by expressing numerous long non-coding RNAs (lncRNAs).
- The functional roles of these HCMV-encoded lncRNAs during the viral life cycle remain largely uncharacterized.
- Understanding viral lncRNA function is crucial for deciphering host-virus interactions.
Purpose of the Study:
- To investigate the functional significance and molecular characteristics of HCMV-encoded lncRNAs.
- To determine the impact of specific lncRNAs on viral latency and lytic replication.
- To elucidate the post-transcriptional regulation and modification of HCMV lncRNAs.
Main Methods:
- Quantification of viral lncRNA expression across the HCMV life cycle.
- Depletion studies using RNA interference to assess lncRNA function.
- Nanopore direct RNA sequencing for transcriptomic analysis.
- Interactome analysis to identify protein binding partners.
- N6-methyladenosine (m6A) profiling and analysis of m6A machinery interactions.
Main Results:
- HCMV lncRNAs constitute a significant portion (30-60%) of the viral transcriptome throughout infection.
- RNA2.7 and RNA4.9 are critical for maintaining the latent reservoir and promoting lytic replication, respectively.
- HCMV lncRNAs are modified with m6A and interact with m6A reader proteins.
- m6A modification machinery stabilizes viral lncRNAs, contributing to their high abundance.
Conclusions:
- HCMV lncRNAs are essential regulators of viral replication and latency.
- Post-transcriptional modification, particularly m6A, plays a key role in stabilizing and regulating viral lncRNAs.
- This study provides a foundation for understanding viral lncRNA-mediated regulation in HCMV infection.
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