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Journal of Virology, May 2000, p. 4541-4548, Vol. 74, No. 10
Plant Research International, 6700 AA
Wageningen,1 and Wageningen
University, 6709 PD Wageningen,2 The
Netherlands
Received 9 September 1999/Accepted 22 February 2000
Luteoviruses avoid degradation in the hemolymph of their aphid
vector by interacting with a GroEL homolog from the aphid's primary
endosymbiotic bacterium (Buchnera sp.). Mutational
analysis of GroEL from the primary endosymbiont of Myzus
persicae (MpB GroEL) revealed that the amino acids mediating
binding of Potato leafroll virus (PLRV;
Luteoviridae) are located within residues 9 to 19 and 427 to 457 of the N-terminal and C-terminal regions, respectively, of the
discontinuous equatorial domain. Virus overlay assays with a series of
overlapping synthetic decameric peptides and their derivatives
demonstrated that R13, K15, L17, and R18 of the N-terminal region and
R441 and R445 of the C-terminal region of the equatorial domain of
GroEL are critical for PLRV binding. Replacement of R441 and R445 by
alanine in full-length MpB GroEL and in MpB GroEL deletion mutants
reduced but did not abolish PLRV binding. Alanine substitution of
either R13 or K15 eliminated the PLRV-binding capacity of the other and
those of L17 and R18. In the predicted tertiary structure of
GroEL, the determinants mediating virus binding are juxtaposed in the
equatorial plain.
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Identifying the Determinants in the Equatorial Domain of
Buchnera GroEL Implicated in Binding Potato
Leafroll Virus
*
Corresponding author. Mailing address: Plant Research
International, P.O. Box 16, 6700 AA Wageningen, The Netherlands. Phone: 31 317 476141. Fax: 31 317 410113. E-mail:
J.F.J.M.vandenHeuvel{at}plant.wag-ur.nl.
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