Tombusvirus p19 Captures RNase III-Cleaved Double-Stranded RNAs Formed by Overlapping Sense and Antisense Transcripts in Escherichia coli

Author:

Huang Linfeng1234,Deighan Padraig56,Jin Jingmin7,Li Yingxue34,Cheung Hung-Chi34,Lee Elaine12,Mo Shirley S.12,Hoover Heather8,Abubucker Sahar8,Finkel Nancy8,McReynolds Larry7,Hochschild Ann5,Lieberman Judy12

Affiliation:

1. Program in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, Massachusetts, USA

2. Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA

3. Department of Biomedical Sciences, Jockey Club College of Veterinary Medicine and Life Sciences, City University of Hong Kong, Hong Kong SAR, China

4. Biotechnology and Health Centre, City University of Hong Kong Shenzhen Research Institute, Nanshan District, Shenzhen, China

5. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA

6. Department of Biology, Emmanuel College, Boston, Massachusetts, USA

7. Division of RNA Biology, New England Biolabs, Ipswich, Massachusetts, USA

8. Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, USA

Abstract

Antisense transcription is widespread in bacteria. By base pairing with overlapping sense RNAs, antisense RNAs (asRNA) can form double-stranded RNAs (dsRNA), which are cleaved by RNase III, a dsRNA endoribonuclease. The ectopic expression of plant Tombusvirus p19 in Escherichia coli stabilizes ∼21-nucleotide (nt) dsRNA RNase III decay intermediates, which enabled us to characterize otherwise highly unstable asRNA by deep sequencing of p19-captured dsRNA. RNase III-produced small dsRNA were formed at most bacterial genes in the bacterial genome and in a plasmid.

Funder

General Research Fund from Research Grants Council of Hong Kong SAR

GSK-IDI Alliance Fellowship

National Natural Science Foundation of China

Science, Technology and Innovation Commission of Shenzhen Municipality

Publisher

American Society for Microbiology

Subject

Virology,Microbiology

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