Genome segment 6 of Antheraea mylitta cypovirus encodes a structural protein with ATPase activity.
The genome segment 6 (S6) of the 11 double stranded RNA genomes from Antheraea mylitta cypovirus was converted into cDNA, cloned and sequenced. S6 consisted of 1944 nucleotides with an ORF of 607 amino acids and could encode a protein of 68 kDa, termed P68. Motif scan and molecular docking analysis of P68 showed the presence of two cystathionine beta synthase (CBS) domains and ATP binding sites. The ORF of AmCPV S6 was expressed in E. coli as His-tag fusion protein and polyclonal antibody was raised. Immunoblot analysis of virus infected gut cells and purified polyhedra using raised anti-p68 polyclonal antibody showed that S6 encodes a viral structural protein. Fluorescence and ATPase assay of soluble P68 produced in Sf-9 cells via baculovirus expression system showed its ability to bind and cleave ATP. These results suggest that P68 may bind viral RNA through CBS domains and help in replication and transcription through ATP binding and hydrolysis.
Chavali
Venkata R M
VR
Department of Biotechnology, Indian Institute of Technology, Kharagpur, Kharagpur 721302, West Bengal, India.
Madhurantakam
Chaithanya
C
Ghorai
Suvankar
S
Roy
Sobhan
S
Das
Amit K
AK
Ghosh
Ananta K
AK
D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D020134:Catalytic Domain; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D016679:Genome, Viral; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology*; D010802:Phylogeny; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012087:Reoviridae / Q000201:enzymology* / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D015678:Viral Structural Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D015678:Viral Structural Proteins; D000251:Adenosine Triphosphatases