Vincent Méjean

Gram-negative bacteria can also form pellicles.

There is a growing interest in the bacterial pellicle, a biofilm floating at the air-liquid interface. Pellicles have been well studied in the Gram-positive bacterium Bacillus subtilis, but far less in Gram-negative bacteria, where pellicle studies have mostly focused on matrix components rather than on the regulatory cascades involved. Several Gram-negative bacteria, including pathogenic bacteria, have been shown to be able to form a pellicle under static conditions. Here, we summarize the growing body of knowledge about pellicle formation in Gram-negative bacteria, especially about the components of the pellicle matrix. We also propose that the pellicle is a specific biofilm, and that its formation involves particular processes. Since this lifestyle concerns a growing number of bacteria, its properties undoubtedly deserve further investigation.

The cyst-dividing bacterium Ramlibacter tataouinensis TTB310 genome reveals a well-stocked toolbox for adaptation to a desert environment.

Ramlibacter tataouinensis TTB310T (strain TTB310), a betaproteobacterium isolated from a semi-arid region of South Tunisia (Tataouine), is characterized by the presence of both spherical and rod-shaped cells in pure culture. Cell division of strain TTB310 occurs by the binary fission of spherical “cyst-like” cells (“cyst-cyst” division). The rod-shaped cells formed at the periphery of a colony (consisting mainly of cysts) are highly motile and colonize a new environment, where they form a new colony by reversion to cyst-like cells. This unique cell cycle of strain TTB310, with desiccation tolerant cyst-like cells capable of division and desiccation sensitive motile rods capable of dissemination, appears to be a novel adaptation for life in a hot and dry desert environment. In order to gain insights into strain TTB310s underlying genetic repertoire and possible mechanisms responsible for its unusual lifestyle, the genome of strain TTB310 was completely sequenced and subsequently annotated. The complete genome consists of a single circular chromosome of 4,070,194 bp with an average G+C content of 70.0%, the highest among the Betaproteobacteria sequenced to date, with total of 3,899 predicted coding sequences covering 92% of the genome. We found that strain TTB310 has developed a highly complex network of two-component systems, which may utilize responses to light and perhaps a rudimentary circadian hourglass to anticipate water availability at the dew time in the middle/end of the desert winter nights and thus direct the growth window to cyclic water availability times. Other interesting features of the strain TTB310 genome that appear to be important for desiccation tolerance, including intermediary metabolism compounds such as trehalose or polyhydroxyalkanoate, and signal transduction pathways, are presented and discussed.

Hsp90 Is Essential under Heat Stress in the Bacterium Shewanella oneidensis

The Hsp90 chaperone is essential in eukaryotes andxa0activates a large array of client proteins. In contrast, its role is still elusive in bacteria, and only a few Hsp90 bacterial clients are known. Here, we found that Hsp90 is essential in the model bacterium Shewanella oneidensis under heat stress. A genetic screen for Hsp90 client proteins identified TilS, anxa0essential protein involved in tRNA maturation. Overexpression of TilS rescued the growth defect of the hsp90 deletion strain under heat stress. Inxa0vivo,xa0the activity and the amount of TilS were significantly reduced in the absence of Hsp90 at high temperature. Furthermore, we showed that Hsp90 interacts with TilS, and Hsp90 prevents TilS aggregation inxa0vitro at high temperature. Together, our results indicate that TilS is a client of Hsp90 in S.xa0oneidensis. Therefore, our study links the essentiality of bacterial Hsp90 at high temperature with the identification of a client.

Aerotaxis governs floating biofilm formation in Shewanella oneidensis

Floating biofilm, or pellicle, is a biofilm found at the air-liquid interface. Here, we show that pellicle development of the aquatic bacterium Shewanella oneidensis is under the control of the chemotaxis system (Che3), a regulatory system known to pilot planktonic cell motion according to environmental cues. Deletion of the histidine kinase cheA3 or the response regulator cheY3 gene led to a heterogeneous pellicle or to the absence of pellicle respectively. In addition, a non-phosphorylatable CheY3-D56A mutant was unable to promote pellicle formation. Kinetic analysis revealed that pellicle formation occurs in three steps: rapid formation of a thin pellicle evolving into a heterogeneous biofilm and finally into a thick homogeneous biofilm. Depletion of oxygen not only abolished initiation of pellicle formation but also blocked pellicle maturation. This study thus demonstrates an essential role of aerotaxis (chemotaxis towards oxygen) in floating biofilm development in S. oneidensis, and it also reveals that pellicle formation is a step-by-step process.

The chemical-in-μwell: a high-throughput technique for identifying solutes eliciting a chemotactic response in motile bacteria.

Bacteria, and in particular marine bacteria, can be found in environments that are poor in nutrients. To survive, they are able to move toward more favorable niches by a mechanism called chemotaxis, whose first step consists in the detection of substrates by chemoreceptors. We developed a chemotactic assay enabling rapid testing of several hundred different solutes and we identified several molecules eliciting a chemotactic response from two aquatic Shewanella species. We propose that this assay be used for other bacteria to determine the repertoire of chemotactic molecules, generally not clearly elucidated.

Competition between VanU(G) repressor and VanR(G) activator leads to rheostatic control of vanG vancomycin resistance operon expression.

Enterococcus faecalis BM4518 is resistant to vancomycin by synthesis of peptidoglycan precursors ending in D-alanyl-D-serine. In the chromosomal vanG locus, transcription of the resistance genes from the PYG resistance promoter is inducible and, upstream from these genes, there is an unusual three-component regulatory system encoded by the vanURSG operon from the PUG regulatory promoter. In contrast to the other van operons in enterococci, the vanG operon possesses the additional vanUG gene which encodes a transcriptional regulator whose role remains unknown. We show by DNase I footprinting, RT-qPCR, and reporter proteins activities that VanUG, but not VanRG, binds to PUG and negatively autoregulates the vanURSG operon and that it also represses PYG where it overlaps with VanRG for binding. In clinical isolate BM4518, the transcription level of the resistance genes was dependent on vancomycin concentration whereas, in a ΔvanUG mutant, resistance was expressed at a maximum level even at low concentrations of the inducer. The binding competition between VanUG and VanRG on the PYG resistance promoter allowed rheostatic activation of the resistance operon depending likely on the level of VanRG phosphorylation by the VanSG sensor. In addition, there was cross-talk between VanSG and VanRG, a VanRG homolog, encoded elsewhere in the chromosome indicating a sophisticated and subtle regulation of vancomycin resistance expression by a complex two-component system.

Effects of a sulfonylurea herbicide on the soil bacterial community

Sulfonylurea herbicides are widely used on a wide range of crops to control weeds. Chevalier® OnePass herbicide is a sulfonylurea herbicide intensively used on cereal crops in Algeria. No information is yet available about the biodegradation of this herbicide or about its effect on the bacterial community of the soil. In this study, we collected an untreated soil sample, and another sample was collected 1xa0month after treatment with the herbicide. Using a high-resolution melting DNA technique, we have shown that treatment with Chevalier® OnePass herbicide only slightly changed the composition of the whole bacterial community. Two hundred fifty-nine macroscopically different clones were isolated from the untreated and treated soil under both aerobic and microaerobic conditions. The strains were identified by sequencing a conserved fragment of the 16S rRNA gene. The phylogenetic trees constructed using the sequencing results confirmed that the bacterial populations were similar in the two soil samples. Species belonging to the Lysinibacillus, Bacillus, Pseudomonas, and Paenibacillus genera were the most abundant species found. Surprisingly, we found that among ten strains isolated from the treated soil, only six were resistant to the herbicide. Furthermore, bacterial overlay experiments showed that only one resistant strain (related to Stenotrophomonas maltophilia) allowed all the sensitive strains tested to grow in the presence of the herbicide. The other resistant strains allowed only certain sensitive strains to grow. On the basis of these results, we propose that there must be several biodegradation pathways for this sulfonylurea herbicide.

YcdY Protein of Escherichia coli, an Atypical Member of the TorD Chaperone Family

The TorD family of specific chaperones is divided into four subfamilies dedicated to molybdoenzyme biogenesis and a fifth one, exemplified by YcdY of Escherichia coli, for which no defined partner has been identified so far. We propose that YcdY is the chaperone of YcdX, a zinc protein involved in the swarming motility process of E. coli, since YcdY interacts with YcdX and increases its activity in vitro.

Efficient respiration on TMAO requires TorD and TorE auxiliary proteins in Shewanella oneidensis.

Respiration on trimethylamine oxide (TMAO) allows bacterial survival under anoxia. In Shewanella oneidensis, Tor is the system involved in TMAO respiration and it is encoded by the torECAD operon. The torA and torC genes encode TorA terminal reductase and the TorC c-type cytochrome, respectively. Sequence analysis suggests that TorD is the putative specific chaperone of TorA, whereas TorE is of unknown function. The purpose of this study was to understand whether TorD and TorE are two accessory proteins that affect the efficiency of the Tor system by chaperoning TorA terminal reductase. Moreover, by deleting each gene, we established that the absence of TorD drastically affects the stability of TorA, while the absence of TorE does not affect TorA stability or activity. Since TMAO reduction was affected in the ΔtorE mutant, TorE could be an additional component of the TorC-TorA electron transfer chain during bacterial respiration. Finally, a fitness experiment indicated that the presence of TorE, as expected, confers a selective advantage in competitive environments.

Dynamic of bacterial communities attached to lightened phytodetritus.

The effects of singlet oxygen (1O2) transfer to bacteria attached on phytodetritus were investigated under laboratory-controlled conditions. For this purpose, a nonaxenic culture of Emiliania huxleyi in late stationary phase was studied for bacterial viability. Our results indicated that only 9u2009±u20093xa0% of attached bacteria were alive compared to 46u2009±u200923xa0% for free bacteria in the E. huxleyi culture. Apparently, under conditions of low irradiance (36xa0W m−2), during the culture, the cumulative dose received (22,000xa0kJ m−2) was sufficiently important to induce an efficient 1O2 transfer to attached bacteria during the senescence of E. huxleyi cells. At this stage, attached bacteria appeared to be dominated by pigmented bacteria (Maribacter, Roseobacter, Roseovarius), which should resist to 1O2 stress probably due to their high contents of carotenoids. After subsequent irradiation of the culture until fully photodegradation of chlorophyll, DGGE analyses showed that the diversity of bacteria attached to E. huxleyi cells is modified by light. Photooxidative alterations of bacteria were confirmed by the increasing amounts of cis-vaccenic photoproducts (bacterial marker) per bacteria observed during irradiation time. Interestingly, preliminary chemotaxis experiments showed that Shewanella oneidensis considered here as a model of motile bacteria was attracted by phytodetritus producing or not 1O2. This lack of repulsive effects could explain the high mortality rate of bacteria measured on E. huxleyi cells.