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Journal of Virology, March 2006, p. 2566-2574, Vol. 80, No. 5
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.5.2566-2574.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Crucial Role of the 5' Conserved Structure of Bamboo Mosaic Virus Satellite RNA in Downregulation of Helper Viral RNA Replication

Yau-Heiu Hsu,1 Hsin-Chuan Chen,2 Jeannie Cheng,2 Padmanaban Annamali,2 Bin-Yen Lin,1,2 Chiang-Tai Wu,2 Wen-Bin Yeh,2,{dagger} and Na-Sheng Lin1,2*

Graduate Institute of Biotechnology, National Chung Hsing University, Taichung 402, Taiwan, Republic of China,1 Institute of Plant and Microbial Biology, Academia Sinica, Taipei 115, Taiwan, Republic of China2

Received 11 July 2005/ Accepted 6 December 2005

Satellite RNA of Bamboo mosaic virus (satBaMV), a single-stranded mRNA type satellite encoding a protein of 20 kDa (P20), depends on the helper BaMV for replication and encapsidation. Two satBaMV isolates, BSF4 and BSL6, exhibit distinctly differential phenotypes in Nicotiana benthamiana plants when coinoculated with BaMV RNA. BSL6 significantly reduces BaMV RNA replication and suppresses the BaMV-induced symptoms, whereas BSF4 does not. By studies with chimeric satBaMVs generated by exchanging the components between BSF4 and BSL6, the genetic determinants responsible for the downregulation of BaMV replication and symptom expression were mapped at the 5' untranslated region (UTR) of BSL6. The 5' UTR of BSL6 alone is sufficient to diminish BaMV RNA replication when the 5' UTR is inserted in cis into the BaMV expression vector or when coinoculation with mutants that block the synthesis of P20 protein takes place. Further, the 5' UTR of natural satBaMV isolates contains one hypervariable (HV) region which folds into a conserved apical hairpin stem-loop (AHSL) structure (W. B. Yeh, Y. H. Hsu, H. C. Chen, and N. S. Lin, Virology 330:105-115, 2004). Interchanges of AHSL segment of HV regions between BSF4 and BSL6 led to the ability of chimeric satBaMV to interfere with BaMV replication and symptom expression. The conserved secondary structure within the HV region is a potent determinant of the downregulation of helper virus replication.

* Corresponding author. Mailing address: Institute of Plant and Microbial Biology, Academia Sinica, Taipei 115, Taiwan, Republic of China. Phone: 886-2-2789-2897. Fax: 886-2-2788-0991. E-mail: nslin{at}sinica.edu.tw.

{dagger} Present address: Department of Entomology, National Chung Hsing University, Taichung 402, Taiwan, Republic of China.

Journal of Virology, March 2006, p. 2566-2574, Vol. 80, No. 5
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.5.2566-2574.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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