Articles
PEANUT BUD NECROSIS VIRUS: PURIFICATION OF NUCLEOCAPSIDS AND SEQUENCE HOMOLOGY OF NUCLEOCAPSID PROTEIN AND GLYCOPROTEIN PRECURSOR WITH OTHER TOSPOVIRUSES
Article number
431_20
Pages
228 – 236
Language
Abstract
A procedure for the purification of peanut bud necrosis virus (PBNV) nucleocapsids was developed.
Virus particles were treated with nonionic detergent to disrupt the envelope membrane and free nucleocapsids were separated into three lightscattering zones after sucrose gradient centrifugation.
Nucleocapsids from the top zone contained S RNA and traces of M RNA; whereas, nucleoplasids from the middle zone contained M RNA with detectable levels of S RNA and those from the bottom zone contained L RNA with traces of M and S RNAs.
Clones from a cDNA library made from the purified PBNV S and M RNAs were characterized and sequenced.
Comparison of the amino-acid sequence of the nucleocapsid (N) protein (276 amino acids) encoded by the S RNA with corresponding sequences from other tospoviruses indicated that PBNV was closely related to members of serogroup IV, i.e., watermelon silver mottle virus (WSMV) and tomato isolate of PBNV (PBNV-To) (identity and similarity values were 86% and 94%, respectively). A more distant relationship was evident between PBNV and members of serogroups I, II, and III (the N protein showed 30–34% identity and 51–53% similarity). Sequence comparisons of the PBNV glycoprotein precursor (GP) protein (1121 amino acids, encoded by the M RNA), also showed distant relationship between PBNV and tomato spotted wilt virus (TSWV) (serogroup I) and impatiens necrotic spot virus (INSV) (serogroup III) (37% identity and 58–59% similarity). These findings suggest that PBNV is a distinct species in serogroup IV.
Virus particles were treated with nonionic detergent to disrupt the envelope membrane and free nucleocapsids were separated into three lightscattering zones after sucrose gradient centrifugation.
Nucleocapsids from the top zone contained S RNA and traces of M RNA; whereas, nucleoplasids from the middle zone contained M RNA with detectable levels of S RNA and those from the bottom zone contained L RNA with traces of M and S RNAs.
Clones from a cDNA library made from the purified PBNV S and M RNAs were characterized and sequenced.
Comparison of the amino-acid sequence of the nucleocapsid (N) protein (276 amino acids) encoded by the S RNA with corresponding sequences from other tospoviruses indicated that PBNV was closely related to members of serogroup IV, i.e., watermelon silver mottle virus (WSMV) and tomato isolate of PBNV (PBNV-To) (identity and similarity values were 86% and 94%, respectively). A more distant relationship was evident between PBNV and members of serogroups I, II, and III (the N protein showed 30–34% identity and 51–53% similarity). Sequence comparisons of the PBNV glycoprotein precursor (GP) protein (1121 amino acids, encoded by the M RNA), also showed distant relationship between PBNV and tomato spotted wilt virus (TSWV) (serogroup I) and impatiens necrotic spot virus (INSV) (serogroup III) (37% identity and 58–59% similarity). These findings suggest that PBNV is a distinct species in serogroup IV.
Authors
T. Satyanarayana, S.E. Mitchell, D.V.R. Reddy, S. Brown, S. Kresovich, R. Jarret, S. Gowda, R.A. Naidu, J.W. Demski
Keywords
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