An antibody to the putative aphid recognition site on cucumber mosaic virus recognizes pentons but not hexons

Valorie D. Bowman, Elaine S. Chase, Alexander W E Franz, Paul R. Chipman, Xing Zhang, Keith L. Perry, Timothy S. Baker, Thomas Smith

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Cucumber mosaic virus (CMV), the type member of the genus Cucumovirus (family Bromoviridae), is transmitted by aphids in a nonpersistent manner. Mutagenesis experiments identified the βH-βI loop of the capsid subunit as a potential key motif responsible for interactions with the insect vector. To further examine the functional characteristics of this motif, we generated monoclonal antibodies that bound to native virions but not to βH-βI mutants. Fab fragments from these antibodies were complexed with wild-type CMV and the virus-Fab structure was determined to 12-Å resolution by using electron cryomicroscopy and image reconstruction techniques. The electron density attributed to the bound antibody has a turret-like appearance and protrudes from each of the 12 fivefold axes of the icosahedral virus. Thus, the antibody binds only to the pentameric clusters (pentons) of A subunits of the T=3 quasisymmetric virus and does not appear to bind to any of the B and C subunits that occur as hexameric clusters (hexons) at the threefold (quasi-sixfold) axes. Modeling and electron density comparisons were used to analyze the paratope-epitope interface and demonstrated that the antibody binds to three βH-βI loops in three adjacent A subunits in each penton. This antibody can discriminate between A and B/C subunits even though the βH-βI loop adopts the same structure in all 180 capsid subunits and is therefore recognizing differences in subunit arrangements. Antibodies with such character have potential use as probes of viral assembly. Our results may provide an additional rationale for designing synthetic vaccines by using symmetrical viral particles.

Original languageEnglish (US)
Pages (from-to)12250-12258
Number of pages9
JournalJournal of Virology
Volume76
Issue number23
DOIs
StatePublished - Dec 2002
Externally publishedYes

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Cucumovirus
Aphids
Cucumber mosaic virus
Aphidoidea
antibodies
Antibodies
capsid
Capsid
virion
Virion
Bromoviridae
synthetic vaccines
electrons
viral morphology
Insect Vectors
Electrons
Antibody Binding Sites
Viruses
Cryoelectron Microscopy
Virus Assembly

ASJC Scopus subject areas

  • Immunology

Cite this

An antibody to the putative aphid recognition site on cucumber mosaic virus recognizes pentons but not hexons. / Bowman, Valorie D.; Chase, Elaine S.; Franz, Alexander W E; Chipman, Paul R.; Zhang, Xing; Perry, Keith L.; Baker, Timothy S.; Smith, Thomas.

In: Journal of Virology, Vol. 76, No. 23, 12.2002, p. 12250-12258.

Research output: Contribution to journalArticle

Bowman, Valorie D. ; Chase, Elaine S. ; Franz, Alexander W E ; Chipman, Paul R. ; Zhang, Xing ; Perry, Keith L. ; Baker, Timothy S. ; Smith, Thomas. / An antibody to the putative aphid recognition site on cucumber mosaic virus recognizes pentons but not hexons. In: Journal of Virology. 2002 ; Vol. 76, No. 23. pp. 12250-12258.
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abstract = "Cucumber mosaic virus (CMV), the type member of the genus Cucumovirus (family Bromoviridae), is transmitted by aphids in a nonpersistent manner. Mutagenesis experiments identified the βH-βI loop of the capsid subunit as a potential key motif responsible for interactions with the insect vector. To further examine the functional characteristics of this motif, we generated monoclonal antibodies that bound to native virions but not to βH-βI mutants. Fab fragments from these antibodies were complexed with wild-type CMV and the virus-Fab structure was determined to 12-{\AA} resolution by using electron cryomicroscopy and image reconstruction techniques. The electron density attributed to the bound antibody has a turret-like appearance and protrudes from each of the 12 fivefold axes of the icosahedral virus. Thus, the antibody binds only to the pentameric clusters (pentons) of A subunits of the T=3 quasisymmetric virus and does not appear to bind to any of the B and C subunits that occur as hexameric clusters (hexons) at the threefold (quasi-sixfold) axes. Modeling and electron density comparisons were used to analyze the paratope-epitope interface and demonstrated that the antibody binds to three βH-βI loops in three adjacent A subunits in each penton. This antibody can discriminate between A and B/C subunits even though the βH-βI loop adopts the same structure in all 180 capsid subunits and is therefore recognizing differences in subunit arrangements. Antibodies with such character have potential use as probes of viral assembly. Our results may provide an additional rationale for designing synthetic vaccines by using symmetrical viral particles.",
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