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Journal of Virology, March 2005, p. 3758-3765, Vol. 79, No. 6
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.6.3758-3765.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Structural Analysis of Tobacco Etch Potyvirus HC-Pro Oligomers Involved in Aphid Transmission{dagger}

Virginia Ruiz-Ferrer,1,{ddagger} Jasminka Boskovic,2,{ddagger} Carlos Alfonso,2 Germán Rivas,2 Oscar Llorca,2 Dionisio López-Abella,1 and Juan José López-Moya1*

Departamento de Biología de Plantas,1 Departamento de Estructura y Función de Proteínas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu 9, Madrid, Spain2

Received 5 August 2004/ Accepted 21 October 2004

Oligomeric forms of the HC-Pro protein of the tobacco etch potyvirus (TEV) have been analyzed by analytical ultracentrifugation and single-particle electron microscopy combined with three-dimensional (3D) reconstruction. Highly purified HC-Pro protein was obtained from plants infected with TEV by using a modified version of the virus that incorporates a histidine tag at the HC-Pro N terminus (hisHC-Pro). The purified protein retained a high biological activity in solution when tested for aphid transmission. Sedimentation equilibrium showed that the hisHC-Pro preparations were heterogenous in size. Sedimentation velocity confirmed the previous observation and revealed that the active protein solution contained several sedimenting species compatible with dimers, tetramers, hexamers, and octamers of the protein. Electron microscopy fields of purified protein showed particles of different sizes and shapes. The reconstructed 3D structures suggested that the observed particles could correspond to dimeric, tetrameric, and hexameric forms of the protein. A model of the interactions required for oligomerization of the HC-Pro of potyviruses is proposed.

* Correspondence author. Mailing address: Departamento de Biología de Plantas, Centro de Investigaciones Biológicas (CIB, CSIC), Ramiro de Maeztu, 9, 28040-Madrid, Spain. Phone: 34-91 837 31 12. Fax: 34-91 536 04 32. E-mail: jjlopez{at}cib.csic.es.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

{ddagger} V.R.-F. and J.B. contributed equally to this work.

Journal of Virology, March 2005, p. 3758-3765, Vol. 79, No. 6
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.6.3758-3765.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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