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Journal of Virology, September 2001, p. 8547-8555, Vol. 75, No. 18
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.18.8547-8555.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Hepatitis Delta Virus Minimal Substrates Competent
for Editing by ADAR1 and ADAR2
Shuji
Sato,1
Swee
Kee
Wong,1,2 and
David W.
Lazinski1,2,*
Department of Molecular Biology and
Microbiology1 and the Raymond and
Beverly Sackler Research Foundation Laboratory, Sackler School of
Graduate Biomedical Sciences,2 Tufts University
School of Medicine, Boston, Massachusetts 02111
Received 12 April 2001/Accepted 5 June 2001
A host-mediated RNA-editing event allows hepatitis delta virus
(HDV) to express two essential proteins, the small delta antigen (HDAg-S) and the large delta antigen (HDAg-L), from a single open reading frame. One or several members of the ADAR (adenosine deaminases that act on RNA) family are thought to convert the adenosine to an
inosine (I) within the HDAg-S amber codon in antigenomic RNA. As a
consequence of replication, the UIG codon is converted to a UGG
(tryptophan [W]) codon in the resulting HDAg-L message. Here, we used
a novel reporter system to monitor the editing of the HDV amber/W site
in the absence of replication. In cultured cells, we observed that both
human ADAR1 (hADAR1) and hADAR2 were capable of editing the amber/W
site with comparable efficiencies. We also defined the minimal HDV
substrate required for hADAR1- and hADAR2-mediated editing. Only 24 nucleotides from the amber/W site were sufficient to enable efficient
editing by hADAR1. Hence, the HDV amber/W site represents the smallest
ADAR substrate yet identified. In contrast, the minimal substrate
competent for hADAR2-mediated editing contained 66 nucleotides.
*
Corresponding author. Mailing address: Department of
Molecular Biology and Microbiology, Tufts University School of
Medicine, 136 Harrison Ave., Boston, MA 02111-1817. Phone: (617)
636-3671. Fax: (617) 636-0337. E-mail:
david.lazinski{at}tufts.edu.
Journal of Virology, September 2001, p. 8547-8555, Vol. 75, No. 18
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.18.8547-8555.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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