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Journal of Virology, January 2001, p. 782-788, Vol. 75, No. 2
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.2.782-788.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Characterization of the AdoMet-Dependent
Guanylyltransferase Activity That Is Associated with the N Terminus of
Bamboo Mosaic Virus Replicase
Yi-Ija
Li,
Yi-Jun
Chen,
Yau-Heiu
Hsu, and
Menghsiao
Meng*
Graduate Institute of Agricultural
Biotechnology, National Chung Hsing University, Taichung, Taiwan
40227, Republic of China
Received 7 February 2000/Accepted 15 October 2000
Bamboo mosaic virus (BaMV), a member of the potexvirus group,
infects primarily members of the Bambusoideae. Open reading frame 1 (ORF1) of BaMV encodes a 155-kDa polypeptide that has long been
postulated to be a replicase involved in the replication and formation
of the cap structure at the 5' end of the viral genome. To identify and
characterize the enzymatic activities associated with the N-terminal
domain of the BaMV ORF1 protein, the intact replicase and two
C-terminally truncated proteins were expressed in Saccharomyces
cerevisiae. All three versions of BaMV ORF1 proteins could be
radiolabeled by [
-32P]GTP, which is a characteristic
of guanylyltransferase activity. The presence of
S-adenosylmethionine (AdoMet) was essential for this
enzymatic activity. Thin-layer chromatography analysis suggests that
the radiolabeled moiety linked to the N-terminal domain of the BaMV
ORF1 protein is m7GMP. The N-terminal domain also exhibited
methyltransferase activity that catalyzes the transfer of the
[3H]methyl group from AdoMet to GTP or
guanylylimidodiphosphate. Therefore, during cap structure formation in
BaMV, methylation of GTP may occur prior to transguanylation as for
alphaviruses and brome mosaic virus. This study establishes the
association of RNA capping activity with the N-terminal domain of the
replicase of potexviruses and further supports the idea that the
reaction sequence of RNA capping is conserved throughout the
alphavirus-like superfamily of RNA viruses.
*
Corresponding author. Mailing address: Graduate
Institute of Agricultural Biotechnology, National Chung Hsing
University, 250 Kuo-Kuang Rd., Taichung, Taiwan 40227, Republic of
China. Phone: 886-4-2840328. Fax: 886-4-2853527. E-mail:
mhmeng{at}dragon.nchu.edu.tw.
Journal of Virology, January 2001, p. 782-788, Vol. 75, No. 2
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.2.782-788.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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