Differentiation of Potyviruses and Their Strains by High Performance Liquid Chromatographic Peptide Profiling of Coat Proteins

Shukla, D. D.; McKern, Neil M.; Gough, K. H.; Tracy, Samantha Lilly; Letho, S. G.

doi:10.1099/0022-1317-69-3-493

Cited by 35 publications

(26 citation statements)

References 16 publications

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“…Potyviruses used in this study were bean yellow mosaic virus (BYMV-S; Randles et al, 1980), clover yellow vein virus (CYVV; Garrett, 1983), Johnson grass mosaic virus (JGMV; Shukla et al, 1987), passion-fruit woodiness virus (PWV-Tip blight; Shukla et al, 1988), potato virus Y (PVY-D; Shukla et al, 1986) and watermelon mosaic virus II (WMMV-II; Greber, 1978). Propagation hosts were Phaseolus vulgaris for BYMV, CYVV and PWV, Zea mays cv.…”

confidence: 99%

The N and C Termini of the Coat Proteins of Potyviruses Are Surface-located and the N Terminus Contains the Major Virus-specific Epitopes

Shukla

Strike

Tracy

et al. 1988

Journal of General Virology

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281

241

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SUMMARYMild proteolysis by trypsin of particles of six potyviruses (bean yellow mosaic virus, clover yellow vein virus, Johnson grass mosaic virus, passion-fruit woodiness virus, potato virus Y and watermelon mosaic virus II) revealed that the N-and C-terminal regions of their coat protein are exposed on the particles' surfaces. The enzyme treatment removed the N-terminal region (30 to 67 amino acids long, depending on the virus) and 18 to 20 amino acids from the C terminus of the coat proteins, leaving a fully assembled virus particle composed of coat protein cores consisting of 216 or 218 amino acid residues. These core particles were indistinguishable from untreated native particles in an electron microscope and were still infectious. The core particles lacked the virus-specific surface epitopes that are recognized by the bulk of the polyclonal antibodies raised against the whole virus particles. Epitopes thought to be groupspecific were located in the trypsin-resistant core protein region. The implications of these findings are discussed in relation to the similar surface location of the N-and Cterminal regions of the coat protein of other rod-shaped plant viruses and the observed common structural features displayed by isometric plant and animal viruses.

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confidence: 99%

The N and C Termini of the Coat Proteins of Potyviruses Are Surface-located and the N Terminus Contains the Major Virus-specific Epitopes

Shukla

Strike

Tracy

et al. 1988

Journal of General Virology

Self Cite

281

241

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“…Although amino acid sequence homology between CPs of potyviruses can differentiate between distinct members and strains, its value as a taxonomic criterion would be greatly enhanced, if this property could be exploited by simpler techniques than sequencing. One approach is the use of high-performance liquid chromatographic peptide profiling which reflects amino acid sequence relationships and readily discrimi nates between viruses and strains [124], This approach has been used in combination with amino acid composition data, partial amino acid sequencing, and immunochemistry [67] to confirm the serological assignment [98] that MDMV-0 is a strain of JGMV and to show that MDMV-KS1 is also a strain of JGMV and is almost identical to MDMV-0 [67]. It is now being employed to resolve the taxonomic relationships of several legumeinfecting potyviruses, including peanut stripe virus, Azuki bean mosaic virus, clover yellow vein virus, pea mosaic virus, BYMV, cowpea aphid-borne mosaic virus, SM V, and other soybean isolates [125].…”

Section: Cp Sequence Datamentioning

confidence: 99%

Taxonomy of Potyviruses: Current Problems and Some Solutions

1991

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There are two major requirements for potyvirus taxonomy; group-specific criteria and criteria which discriminate between distinct viruses and strains. This review discusses the relative merits in potyvirus taxonomy of molecular parameters, such as gene sequence data, nucleic acid hybridization, coat protein sequence data, or high-performance liquid chromatography peptide profiles, and phenotypic characteristics, such as particle morphology, host range, symptomatology, cross-protection, cytoplasmic inclusion morphology, and serology. Coat protein and gene sequence data are the most useful criteria, as they can be used to distinguish viruses from strains and to establish evolutionary relationships between groups of distinct potyviruses. This has led to the revised classification of some viruses and strains and to the clarification of previously conflicting and inconsistent biological properties. It has also provided a sound basis for subgrouping potyviruses. An analysis of the data supports the view that the potyvirus group, including the non-aphid-transmitted viruses, should be elevated to family status, that the vector transmission mode, which correlates with major sequence diversity, should define the four genera, and that distinct potyviruses correspond to species and their variants to strains.

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“…HPLC peptide profiling of potyvi rus coat proteins is a convenient method for checking the validity of such relationships. When used in conjunction with other criteria, as described here and elsewhere [12,19,23,27,37], it can materially assist in the resolution of problems in potyvirus taxonomy.…”

Section: Discussionmentioning

confidence: 99%

“…Solu ble peptides were separated on a Vydac C|S reversephase column (4.6 x 250 mm) in a buffer of 0.1% trifluoroacetic acid, eluted with a linear gradient of 0-35% acetonitrile over 60 min. All other conditions were as described previously [12,23]. The first injection peaks and the last acetonitrile wash peaks were not included in comparisons of profiles.…”

Section: Comparison O F Coat Protein Tryptic Peptides By Hplcmentioning

confidence: 99%

“…In order to assist clarification of the relation ships among some of these legume-infecting potyviruses, we have investigated their coat protein properties, principally by using highperformance liquid chromatographic (HPLC) peptide profiling [12], supported by SDS-PAGE, and amino acid analysis of the coat proteins and derived peptides. The results have been compared with those obtained from coat proteins of some nonlegume infecting potyvi ruses and used as a basis for revision of taxon omic relationships.…”

Section: Introductionmentioning

confidence: 99%

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Coat Protein Properties Suggest that Azuki Bean Mosaic Virus, Blackeye Cowpea Mosaic Virus, Peanut Stripe Virus, and Three Isolates from Soybean Are All Strains of the Same Potyvirus

et al. 1992

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The interrelationship of a number of potyviruses infecting legumes has been investigated by comparing molecular properties of their coat proteins. Comparison of the coat proteins by the techniques of amino acid analysis and PAGE was inadequate to distinguish strains from distinct potyviruses. However, high-performance liquid chromatographic peptide profiles of tryptic digests of coat proteins of these legume-infecting potyviruses enabled such assignments to be made. These data indicate that amino acid sequences of coat proteins of azuki bean mosaic virus, the Type and W strains of blackeye cowpea mosaic virus, three isolates (74, PM, PN) of a potyvirus obtained from soybean in Taiwan, and the Blotch and Mild Mottle strains of peanut stripe virus (PStV) may be very similar to the known sequence of PStV Stripe coat protein. In contrast, peptide profiles of coat proteins from soybean mosaic virus, clover yellow vein virus, bean yellow mosaic virus, potato virus Y, and tobacco etch virus were dissimilar to each other and to the profile of PStV Stripe, suggesting that their coat protein sequences were also quite different. Based on observations of the coat protein structure of many potyviruses, the results suggest that the potyvirus isolates with similar coat proteins are strains of the same potyvirus.

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Differentiation of Potyviruses and Their Strains by High Performance Liquid Chromatographic Peptide Profiling of Coat Proteins

Cited by 35 publications

References 16 publications

The N and C Termini of the Coat Proteins of Potyviruses Are Surface-located and the N Terminus Contains the Major Virus-specific Epitopes

The N and C Termini of the Coat Proteins of Potyviruses Are Surface-located and the N Terminus Contains the Major Virus-specific Epitopes

Taxonomy of Potyviruses: Current Problems and Some Solutions

Coat Protein Properties Suggest that Azuki Bean Mosaic Virus, Blackeye Cowpea Mosaic Virus, Peanut Stripe Virus, and Three Isolates from Soybean Are All Strains of the Same Potyvirus

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