Functional insights from molecular modeling, docking, and dynamics study of a cypoviral RNA dependent RNA polymerase.
Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV) contains 11 double stranded RNA genome segments and infects tasar silkworm A. mylitta. RNA-dependent RNA polymerase (RdRp) is reported as a key enzyme responsible for propagation of the virus in the host cell but its structure function relationship still remains elusive. Here a computational approach has been taken to compare sequence and secondary structure of AmCPV RdRp with other viral RdRps to identify consensus motifs. Then a reliable pairwise sequence alignment of AmCPV RdRp with its closest sequence structure homologue λ3 RdRp is done to predict three dimensional structure of AmCPV RdRp. After comparing with other structurally known viral RdRps, important sequence and/or structural features involved in substrate entry or binding, polymerase reaction and the product release events have been identified. A conserved RNA pentanucleotide (5'-AGAGC-3') at the 3'-end of virus genome is predicted as cis-acting signal for RNA synthesis and its docking and simulation study along with the model of AmCPV RdRp has allowed to predict mode of template binding by the viral polymerase. It is found that template RNA enters into the catalytic center through nine sequence-independent and two sequence-dependent interactions with the specific amino acid residues. However, number of sequence dependent interactions remains almost same during 10 nano-second simulation time while total number of interactions decreases. Further, docking of N(7)-methyl-GpppG (mRNA cap) on the model as well as prediction of RNA secondary structure has shown the template entry process in the active site. These findings have led to postulate the mechanism of RNA-dependent RNA polymerization process by AmCPV RdRp. To our knowledge, this is the first report to evaluate structure function relationship of a cypoviral RdRp.
Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
Anirban
A
Kundu
Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
Anirudha
A
Dutta
Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
Poulomi
P
Biswas
Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
Amit Kumar
AK
Das
Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India. Electronic address: aghosh@hijli.iitkgp.ernet.in.
Ananta Kumar
AK
Ghosh
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D015226:Dinucleoside Phosphates / Q000737:chemistry*; D016679:Genome, Viral; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000737:chemistry*; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry*; D012087:Reoviridae / Q000737:chemistry* / Q000201:enzymology; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D013379:Substrate Specificity; D014764:Viral Proteins / Q000737:chemistry*
D015226:Dinucleoside Phosphates; D012367:RNA, Viral; D014764:Viral Proteins; C062229:7-methyl-diguanosine triphosphate; D012324:RNA-Dependent RNA Polymerase
Antheraea mylitta; Cytoplasmic polyhedrosis virus; Docking and simulation; Double stranded RNA virus; Molecular modeling; RNA dependent RNA polymerase