HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Kass, G. Articles by Loyter, A. Search for Related Content PubMed PubMed Citation Articles by Kass, G. Articles by Loyter, A. Agricola Articles by Kass, G. Articles by Loyter, A.

Permeabilized mammalian cells as an experimental system for nuclear import of geminiviral karyophilic proteins and of synthetic peptides derived from their nuclear localization signal regions

Gideon Kass^1,

Gabriel Arad^1,

¹ Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel
² Department of Plant Genetics, ARO, The Volcani Center, Bet-Dagan 50250, Israel
³ Institut für Molekularbiologie und Biochemie, Free University of Berlin, 14195 Berlin, Germany
⁴ Department of Plant Pathology, University of California, Davis, CA 95616, USA

Correspondence
Abraham Loyter
loyter{at}mail.ls.huji.ac.il

The plant-infecting geminiviruses deliver their genome and viral proteins into the host cell nucleus. Members of the family Geminiviridae possess either a bipartite genome composed of two 2.6 kb DNAs or a monopartite genome of 3.0 kb DNA. The bipartite genome of Bean dwarf mosaic virus (BDMV) encodes several karyophilic proteins, among them the capsid protein (CP) and BV1 (nuclear shuttle protein). A CP is also encoded by the monopartite genome of Tomato yellow leaf curl virus (TYLCV). Here, an in vitro assay system was used for direct demonstration of nuclear import of BDMV BV1 and TYLCV CP, as well as synthetic peptides containing their putative nuclear localization signals (NLSs). Full-length recombinant BDMV BV1 and TYLCV CP mediated import of conjugated fluorescently labelled BSA molecules into nuclei of permeabilized mammalian cells. Fluorescently labelled and biotinylated BSA conjugates bearing the synthetic peptides containing aa 3–20 of TYLCV CP (CP-NLS) or aa 84–106 of BDMV BV1 (BV1-NLS) were also imported into the nuclei of permeabilized cells. This import was blocked by the addition of unlabelled BSA–NLS peptide conjugates or excess unlabelled free NLS peptides. The CP- and BV1-NLS peptides also mediated nuclear import of fluorescently labelled BSA molecules into the nuclei of microinjected mesophyll cells of Nicotiana benthamiana leaves, demonstrating their biological function in intact plant tissue. BV1-NLS and CP-NLS were shown to mediate specific binding to importin , both in vitro and in vivo. These results are consistent with a common nuclear-import pathway for CP and BV1, probably via importin .

These authors contributed equally to this work.

This article has been cited by other articles:

				J. Rosenbluh, Z. Hayouka, S. Loya, A. Levin, A. Armon-Omer, E. Britan, A. Hizi, M. Kotler, A. Friedler, and A. Loyter Interaction between HIV-1 Rev and Integrase Proteins: A BASIS FOR THE DEVELOPMENT OF ANTI-HIV PEPTIDES J. Biol. Chem., May 25, 2007; 282(21): 15743 - 15753. [Abstract] [Full Text] [PDF]