RinA controls phage-mediated packaging and transfer of virulence genes in gram-positive bacteria

Phage-mediated transfer of microbial genetic elements plays a crucial role in bacterial life style and evolution. In this study, we identify the RinA family of phage-encoded proteins as activators required for transcription of the late operon in a large group of temperate staphylococcal phages. RinA binds to a tightly regulated promoter region, situated upstream of the terS gene, that controls expression of the morphogenetic and lysis modules of the phage, activating their transcription. As expected, rinA deletion eliminated formation of functional phage particles and significantly decreased the transfer of phage and pathogenicity island encoded virulence factors. A genetic analysis of the late promoter region showed that a fragment of 272 bp contains both the promoter and the region necessary for activation by RinA. In addition, we demonstrated that RinA is the only phage-encoded protein required for the activation of this promoter region. This region was shown to be divergent among different phages. Consequently, phages with divergent promoter regions carried allelic variants of the RinA protein, which specifically recognize its own promoter sequence. Finally, most Gram-postive bacteria carry bacteriophages encoding RinA homologue proteins. Characterization of several of these proteins demonstrated that control by RinA of the phage-mediated packaging and transfer of virulence factor is a conserved mechanism regulating horizontal gene transfer.

Vertical transmission efficiency and insecticidal properties of Spodoptera exigua Nucleopolyhedrovirus (SeMNPV) genotypes for their use in biological control

El nucleopoliedrovirus de Spodoptera exigua (SeMNPV) es un patógeno natural de las poblaciones larvarias de S. exigua que constituye la base de un bioinsecticida comercializado en España para el control biológico de esta plaga en pimiento. Recientes estudios han demostrado que la transmisión del virus a la descendencia (transmisión vertical) se da con frecuencia y podría ser una característica deseable para su uso en aplicaciones de campo. En el presente trabajo se discute la conveniencia de utilizar una mezcla de dos genotipos SeAl1 (transmisión vertical) y SeG25 (transmisión horizontal) en determinadas proporciones para mejorar las características que cada uno de ellos presenta por separado y así explotar cada una de las vías de transmisión. La patogenicidad (CL50) del genotipo SeG25, y de cualquiera de las mezclas que contienen un 25, 50 o 75 % del mismo, fue más alta que la del aislado SeAl1. Sin embargo, en términos de virulencia (TMM) y productividad (OBs/larva) no se observaron diferencias significativas entre genotipos ni entre sus mezclas. Además se evaluó la capacidad de producir infecciones encubiertas de cada genotipo y sus mezclas sometiendo larvas de S. exigua a infecciones subletales del virus. Se encontraron transcritos del virus para el gen temprano ie0 mediante RT-PCR en los adultos supervivientes a infecciones provocadas por el genotipo SeG25 y todas las mezclas. También se testaron otros dos genes virales que se expresan de manera temprana y tardía en la infección de baculovirus (DNA-polimerasa y polihedrina) para los que en ningún caso se detectaron transcritos.

The extradomain a of fibronectin enhances the efficacy of lipopolysaccharide defective Salmonella bacterins as vaccines in mice

The Extradomain A from fibronectin (EDA) has an immunomodulatory role as fusion protein with viral and tumor antigens, but its effect when administered with bacteria has not been assessed. Here, we investigated the adjuvant effect of EDA in mice immunizations against Salmonella enterica subspecies enterica serovar Enteritidis (Salmonella Enteritidis). Since lipopolysaccharide (LPS) is a major virulence factor and the LPS O-polysaccharide (O-PS) is the immunodominant antigen in serological diagnostic tests, Salmonella mutants lacking O-PS (rough mutants) represent an interesting approach for developing new vaccines and diagnostic tests to differentiate infected and vaccinated animals (DIVA tests). Here, antigenic preparations (hot-saline extracts and formalin-inactivated bacterins) from two Salmonella Enteritidis rough mutants, carrying either intact (SE Delta waaL) or deep-defective (SE Delta gal) LPS-Core, were used in combination with EDA. Biotinylated bacterins, in particular SE Delta waaL bacterin, decorated with EDAvidin (EDA and streptavidin fusion protein) improved the protection conferred by hot-saline or bacterins alone and prevented significantly the virulent infection at least to the levels of live attenuated rough mutants. These findings demonstrate the adjuvant effect of EDAvidin when administered with biotinylated bacterins from Salmonella Enteritidis lacking O-PS and the usefulness of BEDA-SE Delta waaL as non-live vaccine in the mouse model.