CONICET | Buscador de Institutos y Recursos Humanos

In the baculovirus, the homologous regions (hrs) have been implicated both as transcriptional enhancers and origins of DNA replication. Hrs and IBMs (IE1 binding motif) sequences has been shown to bind IE1 protein in gel mobility shift assays. IE1 is an important transcriptional transactivator of the baculovirus. Several studies showed that the IE1 protein has an essential role in the transcriptional regulation of early and late genes and in the viral replication. The goals of this work were to find homologous motif to hrs and IBMs in the AgMNPV genome, and analyze the role in the transcription regulation in presence of the IE1. Computational analysis of the complete AgMNPV genome was used to identified hrs and IBMs motifs similar to that found in AcMNPV. We have identified several putative sequences located upstream to the different ORFs. In order to evaluate if these motifs were involved in the interaction with IE1, we performed gel shift assays, using a DNA fragment containing a putative IBM motif found in the 5´ UTR of ie1 gene. To confirm if these genes were transactivated by IE1, we construct different reporter plasmids with the 5´ UTR region containing the putative hrs or IBMs driving -galactosidase ORF transcription. Through transient assays, we were able to measure b-galactosidase activity in presence or absence of the IE1 and confirm the transactivation.

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