Identification and functional analysis of LsMNPV anti-apoptosis genes

J Biochem Mol Biol. 2007 Jul 31;40(4):571-6. doi: 10.5483/bmbrep.2007.40.4.571.

Abstract

Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING. Ls-P49 contains a reactive site loop, predicted cleavage site (KKLD(74) downward arrow G) that is different from Sl-P49 (TVID(94) downward arrow G). Expressed Ls-iap3 or Ls-p49 under presence of actinomycin D in SF9 cells, DNA ladder assay revealed that Ls- IAP3 or Ls-P49 could block the apoptosis of SF9 cells induced by actinomycin D. Replication of p35 deficient-mutant Autographa californica MNPV in SF9 cells was also rescued when Ls-iap3 or Ls-p49 was expressed transiently. No anti-apoptotic activity was observed for Ls-IAP2. The results showed that both of Ls-IAP3 and Ls-P49 were functional apoptotic suppressors in SF9 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis / drug effects
  • Base Sequence
  • Cell Line
  • DNA Fragmentation / drug effects
  • Dactinomycin / pharmacology
  • Flow Cytometry
  • Inhibitor of Apoptosis Proteins / chemistry
  • Inhibitor of Apoptosis Proteins / genetics*
  • Inhibitor of Apoptosis Proteins / metabolism
  • Molecular Sequence Data
  • Nucleopolyhedroviruses / genetics*
  • Nucleopolyhedroviruses / physiology
  • Sequence Alignment
  • Sequence Analysis, Protein
  • Spodoptera / cytology
  • Spodoptera / drug effects
  • Transfection
  • Viral Proteins / chemistry
  • Viral Proteins / genetics*
  • Viral Proteins / metabolism
  • Virus Replication / drug effects

Substances

  • Inhibitor of Apoptosis Proteins
  • Viral Proteins
  • Dactinomycin