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Journal of Virology, August 2005, p. 9381-9387, Vol. 79, No. 15
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.15.9381-9387.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of Secret Agent as the O-GlcNAc Transferase That Participates in Plum Pox Virus Infection

D. Chen,1 S. Juárez,1 L. Hartweck,2 J. M. Alamillo,1 C. Simón-Mateo,1 J. J. Pérez,1 M. R. Fernández-Fernández,1,{dagger} N. E. Olszewski,2 and J. A. García1*

Department of Plant Molecular Genetics, Centro Nacional de Biotecnología (CSIC), Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain,1 Department of Plant Biology and Plant Molecular Genetics Institute, University of Minnesota, Saint Paul, Minnesota 551082

Received 1 March 2005/ Accepted 23 April 2005

Serine and threonine of many nuclear and cytoplasmic proteins are posttranslationally modified with O-linked N-acetylglucosamine (O-GlcNAc). This modification is made by O-linked N-acetylglucosamine transferases (OGTs). Genetic and biochemical data have demonstrated the existence of two OGTs of Arabidopsis thaliana, SECRET AGENT (SEC) and SPINDLY (SPY), with at least partly overlapping functions, but there is little information on their target proteins. The N terminus of the capsid protein (CP) of Plum pox virus (PPV) isolated from Nicotiana clevelandii is O-GlcNAc modified. We show here that O-GlcNAc modification of PPV CP also takes place in other plant hosts, N. benthamiana and Arabidopsis. PPV was able to infect the Arabidopsis OGT mutants sec-1, sec-2, and spy-3, but at early times of the infection, both rate of virus spread and accumulation were reduced in sec-1 and sec-2 relative to spy-3 and wild-type plants. By matrix-assisted laser desorption ionization-time of flight mass spectrometry, we determined that a 39-residue tryptic peptide from the N terminus of CP of PPV purified from the spy-3 mutant, but not sec-1 or sec-2, was O-GlcNAc modified, suggesting that SEC but not SPY modifies the capsid. While our results indicate that O-GlcNAc modification of PPV CP by SEC is not essential for infection, they show that the modification has a role(s) in the process.

* Corresponding author. Mailing address: Department of Plant Molecular Genetics, Centro Nacional de Biotecnología (CSIC), Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain. Phone: 34 915854535. Fax: 34 915854506. E-mail: jagarcia{at}cnb.uam.es.

{dagger} Present address: Centre for Protein Engineering, MRC Centre, Hills Road, CB2 2QH Cambridge, United Kingdom.

Journal of Virology, August 2005, p. 9381-9387, Vol. 79, No. 15
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.15.9381-9387.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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