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J. Biol. Chem., Vol. 280, Issue 13, 13153-13162, April 1, 2005

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mRNA Guanylation Catalyzed by the S-Adenosylmethionine-dependent Guanylyltransferase of Bamboo Mosaic Virus^*

Yih-Leh Huang, Yau-Heiu Hsu, Yu-Tsung Han, and Menghsiao Meng

From the Graduate Institute of Biotechnology, National Chung Hsing University, 250 Kuo-Kuang Rd., Taichung, Taiwan 40227, Republic of China

The S-adenosylmethionine-dependent guanylyltransferase of bamboo mosaic virus belongs to a novel class of mRNA capping enzymes distantly conserved in Alphavirus-like superfamily. The reaction sequence of the viral enzyme has been proposed comprising steps of 1) binding of GTP and S-adenosylmethionine, 2) formation of m⁷GTP and S-adenosylhomocysteine, 3) formation of the covalent (Enzyme-m⁷GMP) intermediate, and 4) transfer of m⁷GMP from the intermediate to the RNA acceptor. In this study the acceptor specificity of the viral enzyme was characterized. The results show that adenylate or guanylate with 5'-diphosphate group is an essential feature for acceptors, which can be RNA or mononucleotide, to receive m⁷GMP. The transfer rate of m⁷GMP to guanylate is greater than to adenylate by a factor of 3, and the K_m value for mononucleotide acceptor is 10³-fold higher than that for RNA. The capping efficiency of the viral genomic RNA transcript depends on the length of the transcript and the formation of a putative stem-loop structure, suggesting that mRNA capping process may participate in regulating the viral gene expression.

Received for publication, November 8, 2004 , and in revised form, January 24, 2005.

^* This study was supported by National Science Council, Taiwan, Republic of China Grants NSC 92-2313-B-005-071 and NSC 93-2313-B-005-052. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Present address: Institute of Molecular Biology, Academia Sinica, Nankang, Taipei, Taiwan 11529, Republic of China.

To whom correspondence should be addressed. Tel.: 886-422840328; Fax: 886-4-22853527; E-mail: mhmeng{at}dragon.nchu.edu.tw.

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