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Molecular analysis of capsid protein of Homalodisca coagulata Virus-1, a new leafhopper-infecting virus from the glassy-winged sharpshooter, Homalodisca coagulata.

Hunter WB, Katsar CS, Chaparro JX - J. Insect Sci. (2006)

Bottom Line: Sequence identities were analyzed by in silico comparison to known genes in the public database, NCBI.Phylogenetic analysis performed using the optimized protein alignment generated a phylogram containing 5 clades.The HoCV-1, capsid protein sequence has been deposited in GenBank, Accession number: DQ308403.

View Article: PubMed Central - PubMed

Affiliation: United States Department of Agriculture, Agricultural Research Service, US Horticultural Research Laboratory, Fort Pierce, FL 34945, USA. whunter@ushrl.ars.usda.gov

ABSTRACT
A new virus that infects and causes increased mortality in leafhoppers was isolated from the glassy-winged sharpshooter, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). The virus, named Homalodisca coagulata virus -1, HoCV-1, was associated with increased mortality of cultured 5(th) instar H. coagulata. To identify the presence of H. coagulata viral pathogens, cDNA expression libraries were made from adult and nymphs. Analysis using reverse transcriptase PCR demonstrated that the virus was present in midgut tissues. As the viral capsid proteins are commonly used in classification of newly discovered viruses, the capsid proteins (CP) of the virus discovered in H. coagulata was examined. The order of the polyprotein subunits of HoCV-1 capsid proteins was determined to be CP2, CP4, CP3, and CP1. The CP4/CP3 (AFGL/GKPK) cleavage boundary site was clearly identified when the sequences were aligned. The putative CP3/CP1 (ADVQ/SAFA) cleavage site and the putative CP2/CP4 (VTMQ/EQSA) cleavage site of HoCV-1, respectively, were located in the same region as that of the other viruses. After alignment, the CP3/CP1 cleavage sites and CP2/CP4 cleavage sites of the viruses analyzed fell within 50 amino acids of one another. As with the cricket paralysis virus, HoCV-1 was found to be mainly comprised of beta-sandwiches in CP1-3 with a jelly roll topological motif. CP4 of HoCV-1 appeared to be mainly alpha-helical in structure. CP1-4 domains are most homologous to insect picorna-like virus coat proteins as was demonstrated by the results of the BLASTP and PSI-BLAST tests, and is strongly supported by the structural modeling. While sequence homology between the cricket paralysis virus and HoCV-1 was low, the global structure of the proteins was conserved. Sequence identities were analyzed by in silico comparison to known genes in the public database, NCBI. Phylogenetic analysis performed using the optimized protein alignment generated a phylogram containing 5 clades. Clade 1 consisted of Drosophila C virus, Clade 2 consisted of cricket paralysis virus, Clade 3 of Triatoma virus, Plautia stali intestine virus, Himetobi P virus, black queen cell virus, and HoCV-1. Clade 4 encompassed acute bee paralysis virus and Kashmir bee virus, and Clade 5 consisted of Rhopalosiphum padi virus. Analysis of the capsid protein of this new leafhopper virus provided significant evidence that it is related to other ssRNA insect viruses within the Family, Dicistroviridae. The HoCV-1, capsid protein sequence has been deposited in GenBank, Accession number: DQ308403.

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Neighbor-joining analyses of Homalodisca coagulata virus-1, HoCV-1 capsid protein amino acid sequence with selected Cripavirus members: Drosophila C virus (DCV) (Johnson and Christian 1998), cricket paralysis virus (CrPV) (Koonin and Gorbalenya 1992), Triatoma virus (TrV) (Czibener et al. 2000), Plautia stali virus (PSIV) (Sasaki et al. 1998), Himetobi P virus (HiPV) (Nakashima et al. 1999), black queen cell virus (BQCV) (Leat et al. 2000), acute bee paralysis virus (ABPV) (Govan et al. 2000), Kashmir bee virus (KBV)( De Miranda et al. 2004), Rhopalosiphum padi virus (RhPV) (Moon et al. 1998). Phylogenetic trees were constructed via the neighbor-joining method using PAUP* version 4.0 (Swofford 2003). For each tree, confidence levels were estimated using the bootstrap resampling procedure (2000 replications).
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i1536-2442-6-28-1-f03: Neighbor-joining analyses of Homalodisca coagulata virus-1, HoCV-1 capsid protein amino acid sequence with selected Cripavirus members: Drosophila C virus (DCV) (Johnson and Christian 1998), cricket paralysis virus (CrPV) (Koonin and Gorbalenya 1992), Triatoma virus (TrV) (Czibener et al. 2000), Plautia stali virus (PSIV) (Sasaki et al. 1998), Himetobi P virus (HiPV) (Nakashima et al. 1999), black queen cell virus (BQCV) (Leat et al. 2000), acute bee paralysis virus (ABPV) (Govan et al. 2000), Kashmir bee virus (KBV)( De Miranda et al. 2004), Rhopalosiphum padi virus (RhPV) (Moon et al. 1998). Phylogenetic trees were constructed via the neighbor-joining method using PAUP* version 4.0 (Swofford 2003). For each tree, confidence levels were estimated using the bootstrap resampling procedure (2000 replications).

Mentions: The phylogenetic analysis performed using the optimized protein alignment generated a phylogram containing 5 clades (Fig. 3). Clade 1 consisted of Drosophila C virus, Clade 2 consisted of c ricket paralysis virus, Clade 3 of Triatoma virus, Plautia stali intestine virus, Himetobi P virus, black queen cell virus, and HoCV-1. Clade 4 encompassed Acute bee paralysis virus and Kashmir bee virus, and Clade 5 consisted of Rhopalosiphum padi virus. When the four capsid polyprotein units were analyzed individually HoCV-1 always fell within the same clade (Fig. 3) as Triatoma virus, Plautia stali intestine virus, Himetobi P virus, and black queen cell virus.


Molecular analysis of capsid protein of Homalodisca coagulata Virus-1, a new leafhopper-infecting virus from the glassy-winged sharpshooter, Homalodisca coagulata.

Hunter WB, Katsar CS, Chaparro JX - J. Insect Sci. (2006)

Neighbor-joining analyses of Homalodisca coagulata virus-1, HoCV-1 capsid protein amino acid sequence with selected Cripavirus members: Drosophila C virus (DCV) (Johnson and Christian 1998), cricket paralysis virus (CrPV) (Koonin and Gorbalenya 1992), Triatoma virus (TrV) (Czibener et al. 2000), Plautia stali virus (PSIV) (Sasaki et al. 1998), Himetobi P virus (HiPV) (Nakashima et al. 1999), black queen cell virus (BQCV) (Leat et al. 2000), acute bee paralysis virus (ABPV) (Govan et al. 2000), Kashmir bee virus (KBV)( De Miranda et al. 2004), Rhopalosiphum padi virus (RhPV) (Moon et al. 1998). Phylogenetic trees were constructed via the neighbor-joining method using PAUP* version 4.0 (Swofford 2003). For each tree, confidence levels were estimated using the bootstrap resampling procedure (2000 replications).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2990318&req=5

i1536-2442-6-28-1-f03: Neighbor-joining analyses of Homalodisca coagulata virus-1, HoCV-1 capsid protein amino acid sequence with selected Cripavirus members: Drosophila C virus (DCV) (Johnson and Christian 1998), cricket paralysis virus (CrPV) (Koonin and Gorbalenya 1992), Triatoma virus (TrV) (Czibener et al. 2000), Plautia stali virus (PSIV) (Sasaki et al. 1998), Himetobi P virus (HiPV) (Nakashima et al. 1999), black queen cell virus (BQCV) (Leat et al. 2000), acute bee paralysis virus (ABPV) (Govan et al. 2000), Kashmir bee virus (KBV)( De Miranda et al. 2004), Rhopalosiphum padi virus (RhPV) (Moon et al. 1998). Phylogenetic trees were constructed via the neighbor-joining method using PAUP* version 4.0 (Swofford 2003). For each tree, confidence levels were estimated using the bootstrap resampling procedure (2000 replications).
Mentions: The phylogenetic analysis performed using the optimized protein alignment generated a phylogram containing 5 clades (Fig. 3). Clade 1 consisted of Drosophila C virus, Clade 2 consisted of c ricket paralysis virus, Clade 3 of Triatoma virus, Plautia stali intestine virus, Himetobi P virus, black queen cell virus, and HoCV-1. Clade 4 encompassed Acute bee paralysis virus and Kashmir bee virus, and Clade 5 consisted of Rhopalosiphum padi virus. When the four capsid polyprotein units were analyzed individually HoCV-1 always fell within the same clade (Fig. 3) as Triatoma virus, Plautia stali intestine virus, Himetobi P virus, and black queen cell virus.

Bottom Line: Sequence identities were analyzed by in silico comparison to known genes in the public database, NCBI.Phylogenetic analysis performed using the optimized protein alignment generated a phylogram containing 5 clades.The HoCV-1, capsid protein sequence has been deposited in GenBank, Accession number: DQ308403.

View Article: PubMed Central - PubMed

Affiliation: United States Department of Agriculture, Agricultural Research Service, US Horticultural Research Laboratory, Fort Pierce, FL 34945, USA. whunter@ushrl.ars.usda.gov

ABSTRACT
A new virus that infects and causes increased mortality in leafhoppers was isolated from the glassy-winged sharpshooter, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). The virus, named Homalodisca coagulata virus -1, HoCV-1, was associated with increased mortality of cultured 5(th) instar H. coagulata. To identify the presence of H. coagulata viral pathogens, cDNA expression libraries were made from adult and nymphs. Analysis using reverse transcriptase PCR demonstrated that the virus was present in midgut tissues. As the viral capsid proteins are commonly used in classification of newly discovered viruses, the capsid proteins (CP) of the virus discovered in H. coagulata was examined. The order of the polyprotein subunits of HoCV-1 capsid proteins was determined to be CP2, CP4, CP3, and CP1. The CP4/CP3 (AFGL/GKPK) cleavage boundary site was clearly identified when the sequences were aligned. The putative CP3/CP1 (ADVQ/SAFA) cleavage site and the putative CP2/CP4 (VTMQ/EQSA) cleavage site of HoCV-1, respectively, were located in the same region as that of the other viruses. After alignment, the CP3/CP1 cleavage sites and CP2/CP4 cleavage sites of the viruses analyzed fell within 50 amino acids of one another. As with the cricket paralysis virus, HoCV-1 was found to be mainly comprised of beta-sandwiches in CP1-3 with a jelly roll topological motif. CP4 of HoCV-1 appeared to be mainly alpha-helical in structure. CP1-4 domains are most homologous to insect picorna-like virus coat proteins as was demonstrated by the results of the BLASTP and PSI-BLAST tests, and is strongly supported by the structural modeling. While sequence homology between the cricket paralysis virus and HoCV-1 was low, the global structure of the proteins was conserved. Sequence identities were analyzed by in silico comparison to known genes in the public database, NCBI. Phylogenetic analysis performed using the optimized protein alignment generated a phylogram containing 5 clades. Clade 1 consisted of Drosophila C virus, Clade 2 consisted of cricket paralysis virus, Clade 3 of Triatoma virus, Plautia stali intestine virus, Himetobi P virus, black queen cell virus, and HoCV-1. Clade 4 encompassed acute bee paralysis virus and Kashmir bee virus, and Clade 5 consisted of Rhopalosiphum padi virus. Analysis of the capsid protein of this new leafhopper virus provided significant evidence that it is related to other ssRNA insect viruses within the Family, Dicistroviridae. The HoCV-1, capsid protein sequence has been deposited in GenBank, Accession number: DQ308403.

Show MeSH
Related in: MedlinePlus