TY - JOUR
T1 - Turnip yellow mosaic virus isolates with experimentally produced recombinant virion proteins
AU - Hayden, Catherine M.
AU - Mackenzie, Anne M.
AU - Skotnicki, Mary L.
AU - Gibbs, Adrian
PY - 1998/2
Y1 - 1998/2
N2 - The consensus secondary structure of the virion proteins (VPs) of tymoviruses was predicted from their amino acid sequences using a combination of computer methods: profile alignment, hydrophobicity moment and 'PredictProtein'. All methods predicted that they were eight-stranded anti-parallel β-barrels with two α-helical regions. The predicted structure was used to design recombinants of turnip yellow mosaic virus (TYMV) in which selected parts of its VP were replaced with homologous regions of belladonna mottle virus (BeMV) in a cDNA clone encoding the genome of TYMV. Six of ten such recombinants were fully viable and most gave symptoms in Chinese cabbage indistinguishable from those of TYMV, although they did not always infect plants systemically and none infected hosts of BeMV or of other tymoviruses. A TYMV recombinant with the N-terminal part of its VP replaced with the E71 epitope of Plasmodium falciparum was also viable, but others with the same region replaced with the V3 region of the lentivirus human immunodeficiency virus type 1 were not. Epitope analysis of antisera prepared against the virions of parental TYMV and some of the recombinants showed that, although the N terminus of the VP is immunogenically dominant, it is not exposed at the surface of the virion, a finding confirmed by comparing the electrophoretic mobilities of the virions. The recently published structure of the TYMV VP determined by X-ray crystallography confirms the accuracy of the predicted secondary structure of the VP, and hence the utility of the methods used.
AB - The consensus secondary structure of the virion proteins (VPs) of tymoviruses was predicted from their amino acid sequences using a combination of computer methods: profile alignment, hydrophobicity moment and 'PredictProtein'. All methods predicted that they were eight-stranded anti-parallel β-barrels with two α-helical regions. The predicted structure was used to design recombinants of turnip yellow mosaic virus (TYMV) in which selected parts of its VP were replaced with homologous regions of belladonna mottle virus (BeMV) in a cDNA clone encoding the genome of TYMV. Six of ten such recombinants were fully viable and most gave symptoms in Chinese cabbage indistinguishable from those of TYMV, although they did not always infect plants systemically and none infected hosts of BeMV or of other tymoviruses. A TYMV recombinant with the N-terminal part of its VP replaced with the E71 epitope of Plasmodium falciparum was also viable, but others with the same region replaced with the V3 region of the lentivirus human immunodeficiency virus type 1 were not. Epitope analysis of antisera prepared against the virions of parental TYMV and some of the recombinants showed that, although the N terminus of the VP is immunogenically dominant, it is not exposed at the surface of the virion, a finding confirmed by comparing the electrophoretic mobilities of the virions. The recently published structure of the TYMV VP determined by X-ray crystallography confirms the accuracy of the predicted secondary structure of the VP, and hence the utility of the methods used.
UR - http://www.scopus.com/inward/record.url?scp=0031984768&partnerID=8YFLogxK
U2 - 10.1099/0022-1317-79-2-395
DO - 10.1099/0022-1317-79-2-395
M3 - Article
SN - 0022-1317
VL - 79
SP - 395
EP - 403
JO - Journal of General Virology
JF - Journal of General Virology
IS - 2
ER -