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Journal of Virology, September 2003, p. 9685-9694, Vol. 77, No. 17
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.17.9685-9694.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Bromodomain-Containing Protein from Tomato Specifically Binds Potato Spindle Tuber Viroid RNA In Vitro and In Vivo

Angel Emilio Martínez de Alba,1,{dagger} Rudolf Sägesser,1 Martin Tabler,1 and Mina Tsagris1,2*

Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology,1 Department of Biology, University of Crete, Heraklion, Crete, Greece2

Received 25 February 2003/ Accepted 27 May 2003

For the identification of RNA-binding proteins that specifically interact with potato spindle tuber viroid (PSTVd), we subjected a tomato cDNA expression library prepared from viroid-infected leaves to an RNA ligand screening procedure. We repeatedly identified cDNA clones that expressed a protein of 602 amino acids. The protein contains a bromodomain and was termed viroid RNA-binding protein 1 (VIRP1). The specificity of interaction of VIRP1 with viroid RNA was studied by different methodologies, which included Northwestern blotting, plaque lift, and electrophoretic mobility shift assays. VIRP1 interacted strongly and specifically with monomeric and oligomeric PSTVd positive-strand RNA transcripts. Other RNAs, for example, U1 RNA, did not bind to VIRP1. Further, we could immunoprecipitate complexes from infected tomato leaves that contained VIRP1 and viroid RNA in vivo. Analysis of the protein sequence revealed that VIRP1 is a member of a newly identified family of transcriptional regulators associated with chromatin remodeling. VIRP1 is the first member of this family of proteins, for which a specific RNA-binding activity is shown. A possible role of VIRP1 in viroid replication and in RNA mediated chromatin remodeling is discussed.

* Corresponding author. Mailing address: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, P.O. Box 1527, 71110 Heraklion, Greece. Phone: 30-2810-394367. Fax: 30-2810-394408. E-mail: tsagris{at}imbb.forth.gr.

{dagger} Present address: Instituto de Biologia Molecular y Cellular de Plantas, 46022 Valencia, Spain.

Journal of Virology, September 2003, p. 9685-9694, Vol. 77, No. 17
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.17.9685-9694.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kalantidis, K., Denti, M. A., Tzortzakaki, S., Marinou, E., Tabler, M., Tsagris, M. (2007). Virp1 Is a Host Protein with a Major Role in Potato Spindle Tuber Viroid Infection in Nicotiana Plants. J. Virol. 81: 12872-12880 [Abstract] [Full Text]  
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