Prophage maintenance into enterobacterial genomes

Les bactériophages sont les virus spécifiques des bactéries. Ils sont considérés comme les entités biologiques les plus abondantes de la biosphère (1031 au total). Une grande partie des bactériophages sont dits tempérés de part leur propriété à intégrer leur génome dans celui de leur hôte et à s'y maintenir en état de réplication passive appelé lysogénie. Les gènes de prophages apportent de nouvelles propriétés à l'hôte via la conversion lysogénique. De nombreux prophages défectifs et fonctionnels sont maintenus dans les génomes bactériens. Nous avons émis l'hypothèse que des stratégies de maintien aient été sélectionnées pour maintenir cette source de gènes, même si elle est potentiellement dangereuse car les prophages peuvent être induits dans des conditions de stress.Nos résultats suggèrent que le maintien de la lysogénie d'une catégorie de prophages, qui présente une organisation génétique atypique du module de recombinaison spécifique de site, est sous le contrôle du facteur de terminaison de la transcription Rho. Pour ces prophages, qu'ils soient défectifs ou fonctionnels, leur induction par inactivation de Rho, fait intervenir une nouvelle voie d'induction lytique indépendante de la voie classique via la réponse SOS.Ces interactions hôtes-virus reflète la coévolution de ces microorganismes, qui permet l'acquisition de gènes via le transfert horizontal tout en contrôlant l'expression des gènes délétères. Ceci permet l'acquisition de nouvelles propriétés et l'adaptation de l'hôte à différentes conditions environnementales.Bacteriophages are the most abundant biological entities in the biosphere. A majority of them are temperate phages that are able to integrate their genome into the host and replicate passively in a lysogenic state. Hosts frequently benefit from such massive gene acquisition through lysogenic conversion. As prophages may be beneficial to their hosts, we hypothesize that hosts adapted strategies for maintaining that gene source. Since prophages integrate into and excise from the host chromosome through site-specific recombination (SSR), we investigated whether regulation of SSR at the level of gene expression could be involved in the maintenance process. Our results suggest that lysogeny maintenance of a class of prophages, which all share a same unusual genetic organization, are controlled by the transcription termination factor Rho. Rho is not only involved in horizontally acquired gene silencing but also in prophage maintenance, which can be seen as an adaptation of the host to maintain prophage genes. For these prophages, whether defective or functional, their induction by the inactivation of Rho, involves a new pathway of lysogeny escape, which is independent of the classical pathway via the SOS response. This newly characterized interaction reflects the coevolution of host and viruses, which allows the acquisition of genes, and thus new properties, via horizontal transfer, while controlling the expression of deleterious genes

Subspace Analysis of Indoor UWB Channels

This work aims at characterizing the second-order statistics of indoor ultra-wideband (UWB) channels using channel sounding techniques. We present measurement results for different scenarios conducted in a laboratory setting at Institut Eurécom. These are based on an eigendecomposition of the channel autocovariance matrix, which allows for the analysis of the growth in the number of significant degrees of freedom of the channel process as a function of the signaling bandwidth as well as the statistical correlation between different propagation paths. We show empirical eigenvalue distributions as a function of the signal bandwidth for both line-of-sight and non-line-of-sight situations. Furthermore, we give examples where paths from different propagation clusters (possibly arising from reflection or diffraction) show strong statistical dependence

Bacterial genome remodeling through bacteriophage recombination

Bacteriophages co-exist and co-evolve with their hosts in natural environments. Virulent phages lyse infected cells through lytic cycles, whereas temperate phages often remain dormant and can undergo lysogenic or lytic cycles. In their lysogenic state, prophages are actually part of the host genome and replicate passively in rhythm with host division. However, prophages are far from being passive residents: they can modify or bring new properties to their host. In this review, we focus on two important phage-encoded recombination mechanisms, i.e. site-specific recombination and homologous recombination, and how they remodel bacterial genomes

Transcription termination controls prophage maintenance in Escherichia coli genomes.

International audienceProphages represent a large fraction of prokaryotic genomes and often provide new functions to their hosts, in particular virulence and fitness. How prokaryotic cells maintain such gene providers is central for understanding bacterial genome evolution by horizontal transfer. Prophage excision occurs through site-specific recombination mediated by a prophage-encoded integrase. In addition, a recombination directionality factor (or excisionase) directs the reaction toward excision and prevents the phage genome from being reintegrated. In this work, we describe the role of the transcription termination factor Rho in prophage maintenance through control of the synthesis of transcripts that mediate recombination directionality factor expression and, thus, excisive recombination. We show that Rho inhibition by bicyclomycin allows for the expression of prophage genes that lead to excisive recombination. Thus, besides its role in the silencing of horizontally acquired genes, Rho also maintains lysogeny of defective and functional prophages

Chaperone-assisted Excisive Recombination, a Solitary Role for DnaJ (Hsp40) Chaperone in Lysogeny Escape

International audienceTemperate bacteriophage lytic development is intrinsically related to the stress response in particular at the DNA replication and virion maturation steps. Alternatively, temperate phages become lysogenic and integrate their genome into the host chromosome. Under stressful conditions, the prophage resumes a lytic development program, and the phage DNA is excised before being replicated. The KplE1 defective prophage of Escherichia coli K12 constitutes a model system because it is fully competent for integrative as well as excisive recombination and presents an atypical recombination module, which is conserved in various phage genomes. In this work, we identified the host-encoded stress-responsive molecular chaperone DnaJ (Hsp40) as an active participant in KplE1 prophage excision. We first show that the recombination directionality factor TorI of KplE1 specifically interacts with DnaJ. In addition, we found that DnaJ dramatically enhances both TorI binding to its DNA target and excisive recombination in vitro. Remarkably, such stimulatory effect by DnaJ was performed independently of its DnaK chaperone partner and did not requirea functional DnaJ J-domain. Taken together, our results underline a novel and unsuspected functional interaction between the generic host stress-regulated chaperone and temperate bacteriophage lysogenic development

Retrotransposons. An RNA polymerase III subunit determines sites of retrotransposon integration.

International audienceMobile genetic elements are ubiquitous. Their integration site influences genome stability and gene expression. The Ty1 retrotransposon of the yeast Saccharomyces cerevisiae integrates upstream of RNA polymerase III (Pol III)-transcribed genes, yet the primary determinant of target specificity has remained elusive. Here we describe an interaction between Ty1 integrase and the AC40 subunit of Pol III and demonstrate that AC40 is the predominant determinant targeting Ty1 integration upstream of Pol III-transcribed genes. Lack of an integrase-AC40 interaction dramatically alters target site choice, leading to a redistribution of Ty1 insertions in the genome, mainly to chromosome ends. The mechanism of target specificity allows Ty1 to proliferate and yet minimizes genetic damage to its host

A small targeting domain in Ty1 integrase is sufficient to direct retrotransposon integration upstream of tRNA genes

Integration of transposable elements into the genome is mutagenic. Mechanisms targeting integrations into relatively safe locations, hence minimizing deleterious consequences for cell fitness, have emerged during evolution. In budding yeast, integration of the Ty1 LTR retrotransposon upstream of RNA polymerase III (Pol III)-transcribed genes requires interaction between Ty1 integrase (IN1) and AC40, a subunit common to Pol I and Pol III. Here, we identify the Ty1 targeting domain of IN1 that ensures (i) IN1 binding to Pol I and Pol III through AC40, (ii) IN1 genome-wide recruitment to Pol I- and Pol III-transcribed genes, and (iii) Ty1 integration only at Pol III-transcribed genes, while IN1 recruitment by AC40 is insufficient to target Ty1 integration into Pol I-transcribed genes. Swapping the targeting domains between Ty5 and Ty1 integrases causes Ty5 integration at Pol III-transcribed genes, indicating that the targeting domain of IN1 alone confers Ty1 integration site specificity