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Cylindrical inclusion protein of potato virus A is associated with a subpopulation of particles isolated from infected plants.

Authors :
Gabrenaite-Verkhovskaya R
Andreev IA
Kalinina NO
Torrance L
Taliansky ME
Mäkinen K
Source :
The Journal of general virology [J Gen Virol] 2008 Mar; Vol. 89 (Pt 3), pp. 829-838.
Publication Year :
2008

Abstract

Potato virus A (PVA) particles were purified by centrifugation through a 30 % sucrose cushion and the pellet (P1) was resuspended and sedimented through a 5-40 % sucrose gradient. The gradient separation resulted in two different virus particle populations: a virus fraction (F) that formed a band in the gradient and one that formed a pellet (P2) at the bottom of the gradient. All three preparations contained infectious particles that retained their integrity when visualized by electron microscopy (EM). Western blotting of the P1 particles revealed that the viral RNA helicase, cylindrical inclusion protein (CI), co-purified with virus particles. This result was confirmed with co-immunoprecipitation experiments. CI was detected in P2 particle preparations, whereas F particles were devoid of detectable amounts of CI. ATPase activity was detected in all three preparations with the greatest amount in P2. Results from immunogold-labelling EM experiments suggested that a fraction of the CI present in the preparations was localized to one end of the virion. Atomic force microscopy (AFM) studies showed that P1 and P2 contained intact particles, some of which had a protruding tip structure at one end, whilst F virions were less stable and mostly appeared as beaded structures under the conditions of AFM. The RNA of the particles in F was translated five to ten times more efficiently than RNA from P2 particles when these preparations were subjected to translation in wheat-germ extracts. The results are discussed in the context of a model for CI-mediated functions.

Details

Language :
English
ISSN :
0022-1317
Volume :
89
Issue :
Pt 3
Database :
MEDLINE
Journal :
The Journal of general virology
Publication Type :
Academic Journal
Accession number :
18272775
Full Text :
https://doi.org/10.1099/vir.0.83406-0