Histone H4 is a protein subunit of nucleosomes in eukaryotes and play crucial roles in DNA package and in regulation of gene expression by covalent modification. A viral histone H4 is encoded in a polydnavirus called Cotesia plutellae bracovirus (CpBV). The viral H4 (CpBV-H4) is highly homologous with other H4 proteins except 38 extended residues in N terminus. Its expression alters insect gene expression and suppresses immune and development. In this study, CpBV-H4 was expressed in a natural host, Plutella xylostella, and its suppressive activity on host gene expression was detected by suppressive subtractive hybridization (SSH) technique. SSH targets, of which expressions were down-regulated by CpBV-H4, were read by 454 pyrosequencing and annotated using the published P. xylostella whole genome. Resulting targets were assigned to most GO functional categories. Two chromatin remodeling factors were included in the SSH targets. Lysine demethylase (Px-KDM) of P. xylostella was highly expressed during entire larval period in all tested tissues. However, the suppression of Px-KDM expression by a specific RNA interference (RNAi) did not affect immune response, but significantly impaired the larval development. SWI/SNF of P. xylostella (Px-SWI/SNF) was expressed in all developmental stages. Its RNAi did not affect larval development, but led to significant alteration in adult metamorphosis. CpBV-H4 suppressed expressions of both Px-KDM and Px-SWI/SNF, but its truncated mutant lacking in the extended N-terminal tail did not. These results suggest that the developmental alteration in P. xylostella parasitized by C. plutellae can be caused by an epigenetic control of CpBV-H4 against chromatin remodeling factors.

Cotesia plutellae, an endoparasitoids braconid wasp, possesses a polydnaviruses (PDVs) called Cotesia plutellae bracovirus (CpBV) that encodes viral histone H4 (= CpBV-H4). This viral histone H4 shares high sequence homology (82.5%) with host`s H4 of P.xylostella, except an extended N-terminal tail consisting of 38 amino acid residues with nine lysines. Its extended N-terminal tail has been postulated to play a crucial role in suppressing host immunity, growth and development-associated genes, presumably through an epigenetic control mechanism. A suppression subtractive hybridization (SSH) analysis was analyzed in transcriptome by short-read sequencing technology and provided several target and non-target genes of a viral histone H4. In this study, we analyzed the effect CpBV-H4 on the expression of two target genes: Lysine-specific demethylase (KDM) and Serine proteinase inhibitor (Serpins). Transient expression of CpBV-H4 into non parasitized P. xylostella was performed by microinjection of a recombinant expression vector, and showed the expression up to 70 h. Under this transient expression condition, we analyzed the effect of CpBV-H4 on the expression of target genes by RT-PCR at different time points. Interestingly, the CpBV-H4 significantly inhibited the expression of these target genes, while the truncated CpBV-H4 deleting the N-terminal tail did not show this inhibitory effect. This study also showed that the viral histone H4 suppresses expressions of lysine-specific demethylase and serine proteinase inhibitor (Serpin2) to inhibit host growth and development.