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Volume 97, Issue 7

Reconstitution of the RNA-dependent RNA polymerase activity of cypovirus using separately expressed different functional domains of the enzyme Free

Abstract

cytoplasmic polyhedrosis virus is a segmented dsRNA virus of the family . Segment 2 (S2)-encoded RNA-dependent RNA polymerase (RdRp) helps the virus to propagate its genome in the host cell of the silkworm, . Cloning, expression, purification and functional analysis of individual domains of RdRp have demonstrated that the purified domains interact . The central polymerase domain (PD) shows nucleotide binding properties, but neither the N-terminal domain (NTD) nor the C-terminal domain (CTD). Isolated PD does not exhibit RdRp activity but this activity can be reconstituted when all three domains are included in the reaction mixture. Molecular dynamics simulation suggests that the isolated PD has increased internal motions in comparison to when it is associated with the NTD and CTD. The motions of the separated PD may lead to the formation of a less accessible RNA template-binding channel and, thus, impair RdRp activity.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Antheraea mylitta , cytoplasmic polyhedrosis virus , insect virus , molecular dynamics simulation. , protein domains and RNA-dependent RNA polymerase
© 2016 The Authors
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2016-07-01
2024-04-23
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Reconstitution of the RNA-dependent RNA polymerase activity of Antheraea mylitta cypovirus in vitro using separately expressed different functional domains of the enzyme
J. Gen. Virol. 97, 1709 (2016); https://doi.org/10.1099/jgv.0.000463
/content/journal/jgv/10.1099/jgv.0.000463
/content/journal/jgv/10.1099/jgv.0.000463
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