tailieunhanh - Báo cáo khoa học: Leadzyme formed in vivo interferes with tobacco mosaic virus infection in Nicotiana tabacum

We developed a new method for inhibiting tobacco mosaic virus infection in tobacco plants based on specific RNA hydrolysis induced by a leadzyme. We identified a leadzyme substrate target sequence in genomic tobacco mosaic virus RNA and designed a 16-mer oligoribonucleotide capable of forming a specific leadzyme motif with a five-nucleotide catalytic loop. | ễFEBS Journal Leadzyme formed in vivo interferes with tobacco mosaic virus infection in Nicotiana tabacum Eliza Wyszko1 Monika Nowak1 Henryk Pospieszny2 Maciej Szymanski1 Jakub Pas1 3 Mirostawa Z. Barciszewska1 and Jan Barciszewski1 1 Institute of Bioorganic Chemistry Polish Academy of Sciences Poznan Poland 2 Institute of Plant Protection Department of Virology and Bacteriology Poznan Poland 3 BioInfoBank Institute Poznan Poland Keywords leadzyme RNA catalysis RNA hydrolysis TMV tobacco mosaic virus Correspondence J. Barciszewski Institute of Bioorganic Chemistry of the Polish Academy of Sciences Noskowskiego 12 61-704 Poznan Poland Fax 48 618520532 Tel 48 618528503 E-mail Received 5 June 2006 revised 3 August 2006 accepted 12 September 2006 doi We developed a new method for inhibiting tobacco mosaic virus infection in tobacco plants based on specific RNA hydrolysis induced by a leadzyme. We identified a leadzyme substrate target sequence in genomic tobacco mosaic virus RNA and designed a 16-mer oligoribonucleotide capable of forming a specific leadzyme motif with a five-nucleotide catalytic loop. The synthetic 16-mer RNA was applied with nontoxic catalytic amount of lead to infected tobacco leaves. We observed inhibition of tobacco mosaic virus infection in tobacco leaves in vivo due to specific tobacco mosaic virus RNA cleavage effected by leadzyme. A significant reduction in tobacco mosaic virus accumulation was observed even when the leadzyme was applied up to 2 h after inoculation of leaves with tobacco mosaic virus. This process called leadzyme interference is determined by specific recognition and cleavage of the target site by the RNA catalytic strand in the presence of Pb2 . Catalytic RNAs ribozymes are capable of specific binding and cleaving of RNA molecules. Since their discovery many efforts have been made to explore them as tools for silencing of viral genes and inhibition of viral .