A potential nuclear envelope-targeting domain and an arginine-rich RNA binding element identified in the putative movement protein of the GAV strain of Barley yellow dwarf virus
Zongliang Xia A B E , Yan Wang A B E , Zhiqiang Du A , Junmin Li A C , Richard Y. Zhao D F and Daowen Wang A FA The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
B Graduate School of Chinese Academy of Sciences, Beijing 100039, China.
C The Centre for Agricultural Resources, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
D Departments of Pathology and Microbiology-Immunology, Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, MSTF 600, Baltimore, MD 21201-1192, USA.
E These authors contributed equally to this article.
F Corresponding authors. Emails: dwwang@genetics.ac.cn; rzhao@som.umaryland.edu
Functional Plant Biology 35(1) 40-50 https://doi.org/10.1071/FP07114
Submitted: 3 May 2007 Accepted: 3 December 2007 Published: 25 January 2008
Abstract
In this study, the structural elements in the putative movement protein (MP) of the GAV strain of Barley yellow dwarf virus (BYDV-GAV) were investigated. The GFP fusion protein of BYDV-GAV MP was found to be associated with the nuclear envelope (NE) in transgenic Arabidopsis thaliana (L.) cells. Serial deletion mapping demonstrated that the predicted α-helical domain located at the N-terminus of BYDV-GAV MP was required and sufficient for NE targeting in onion epidermal cells. This α-helical domain does not contain any sequence elements similar to known nuclear localisation signals or bear any significant resemblance to previously characterised NE-targeting structure, indicating that it may represent a novel NE-targeting domain in plant cells. Deletion mutagenesis showed that the C-terminal end of BYDV-GAV MP possessed an element required for its RNA binding activity in vitro. Further analysis revealed that the arginine amino acids within the last 11 residues of the C-terminal end were crucial for the binding of BYDV-GAV MP to RNA. This C-terminal element enriched in basic residues was also present in the MPs of other BYDV strains and the polerovirus Potato leaf roll virus (PLRV), suggesting the conservation of a RNA binding element in the MPs from both luteoviruses and poleroviruses. The data in this work present an initial characterisation of a novel plant NE-targeting domain and a RNA binding element on BYDV-GAV MP. Further studies are underway to investigate the function of these elements in the biology of natural BYDV-GAV infection.
Additional keywords: Arabidopsis thaliana, luteovirus.
Acknowledgements
We thank Professor Nam-Hai Chua (The Rockefeller University, USA) and Dr Jianru Zuo (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, China) for providing the XVE construct, and Professor David Phoenix and Dr Frederick Harris (Department of Forensic and Investigative Science, University of Central Lancashire, UK) for helpful discussions on this work. In addition, we are grateful to Drs Sylvian Huard (University of Maryland, USA) and David Leader (Scottish Crop Research Institute, Scotland) for correcting the manuscript and to the reviewers for constructive suggestions on manuscript revision. Financial support for this work is provided by the National Natural Science Foundation of China (30521001) and the Ministry of Science and Technology of China (G2000016201).
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