Gre factors are required fot biofilm formation in Salmonella enterica serovar Typhimurium by targeting transcription of the cgsD gene

Rdar biofilm formation of Salmonella typhimurium and Escherichia coli is a common ancient multicellular behavior relevant in cell-cell and inter-organism interactions equally, as in interaction with biotic and abiotic surfaces. With the expression of the characteristic extracellular matrix components amyloid curli fimbriae and the exopolysaccharide cellulose, the central hub for the delicate regulation of rdar morphotype expression is the orphan transcriptional regulator CsgD. Gre factors are ubiquitously interacting with RNA polymerase to selectively overcome transcriptional pausing. In this work, we found that GreA/GreB are required for expression of the csgD operon and consequently the rdar morphotype. The ability of the Gre factors to suppress transcriptional pausing and the 147 bp 5′-UTR of csgD are required for the stimulatory effect of the Gre factors on csgD expression. These novel mechanism(s) of regulation for the csgD operon might be relevant under specific stress conditions

ppGpp, the general stress response alarmone, is required for the expression of the alpha-hemolysin toxin in the uropathogenic EScherichia coli isolate, J96.

ppGpp is an intracellular sensor that, in response to different types of stress, coordinates the rearrangement of the gene expression pattern of bacteria to promote adaptation and survival to new environmental conditions. First described to modulate metabolic adaptive responses, ppGpp modulates the expression of genes belonging to very diverse functional categories. In Escherichia coli, ppGpp regulates the expression of cellular factors that are important during urinary tract infections. Here, we characterize the role of this alarmone in the regulation of the hlyCABDII operon of the UPEC isolate J96, encoding the toxin α-hemolysin that induces cytotoxicity during infection of bladder epithelial cells. ppGpp is required for the expression of the α-hemolysin encoded in hlyCABDII by stimulating its transcriptional expression. Prototrophy suppressor mutations in a ppGpp-deficient strain restore the α-hemolysin expression from this operon to wild-type levels, confirming the requirement of ppGpp for its expression. ppGpp stimulates hlyCABDII expression independently of RpoS, RfaH, Zur, and H-NS. The expression of hlyCABDII is promoted at 37 °C and at low osmolarity. ppGpp is required for the thermoregulation but not for the osmoregulation of the hlyCABDII operon. Studies in both commensal and UPEC isolates demonstrate that no UPEC specific factor is strictly required for the ppGpp-mediated regulation described. Our data further support the role of ppGpp participating in the coordinated regulation of the expression of bacterial factors required during infection

Patógenos de importancia clínica

PublishedEl presente libro Patógenos de importancia clínica: Inves­tigaciones Recientes en el Valle del Cauca nace como una iniciativa para socializar los resultados obtenidos en los tra­bajos de investigación más recientes de los miembros del Grupo de Investigación en genética, fisiología y metabolis­mo –GEFIME– de la Universidad Santiago de Cali. Estas investigaciones involucran patógenos que son importantes agentes causales de infecciones y de alto impacto en la salud pública en la ciudad de Cali y en el Valle del Cauca. Este libro consta de cinco capítulos con trabajos de investiga­ción en bacterias resistentes a los antibióticos como Staphylo­coccus aureus y Mycobacterium tuberculosis y su comporta­miento epidemiológico en ambientes intrahospitalarios y en poblaciones, respectivamente. En el Capítulo 3 se presenta un estudio epidemiológico que establece la presencia del virus del papiloma humano (VPH) en mucosa oral y su relación con el desarrollando cáncer oral y en el Capítulo 4 se presenta una investigación realizada con moléculas efectoras (alarmona pp­Gpp y la proteína DksA) de Salmonella enterica serovar Typhi­murium y su influencia en la patogenicidad y la formación de biopelículas in-vitro

Expression of accessory genes in Salmonella requires the presence of the Gre factors

Genomic studies with Salmonella enterica serovar Typhimurium reveal a crucial role of horizontal gene transfer (HGT) in the acquisition of accessory cellular functions involved in host-interaction. Many virulence genes are located in genomic islands, plasmids and prophages. GreA and GreB proteins, Gre factors, interact transiently with the RNA polymerase alleviating backtracked complexes during transcription elongation. The overall effect of Gre factors depletion in Salmonella expression profile was studied. Both proteins are functionally redundant since only when both Gre factors were depleted a major effect in gene expression was detected. Remarkably, the accessory gene pool is particularly sensitive to the lack of Gre factors, with 18.6% of accessory genes stimulated by the Gre factors versus 4.4% of core genome genes. Gre factors involvement is particularly relevant for the expression of genes located in genomic islands. Our data reveal that Gre factors are required for the expression of accessory genes

First Report of Acanthocheilonema reconditum Outbreak in Canines with Clinical Signs of Anemia from Southwestern Colombia

Different nematodes affect canines, however Acanthocheilonema reconditum was considered mostly a non-pathogenic parasite. Climate change, animal migration, and other factors transformed the dynamics of vector-borne diseases, including filariasis. Since 2016, a sudden increase in the number of dogs with microfilaremia was reported by different veterinary centers in Cali, southwest Colombia. The objective of this study was to molecularly identify the etiologic agent of this filariasis outbreak detected in this city, using PCR–RFLP and evaluating dogs’ clinical signs. From 2018–2019, canine filariasis cases were prospectively evaluated after a microscopic test, recruiting 82 cases and 43 healthy controls from 2971 samples. Acanthocheilonema reconditum (Nematoda, Onchocercidae) was identified in 61.3% of the cases (49/82) by PCR–RFLP. Sanger sequencing of the 5.8S ribosomal RNA gene and internal transcribed spacer-2 fragment was additionally performed on seven cases, confirming A. reconditum in all of them. The filariasis cases are statistically associated with male dogs who have clinical signs of anemia, low levels of hemoglobin and hematocrit (p < 0.0001), and high levels of plasma proteins (p < 0.001). This emerging canine disease constitutes an important public health concern among veterinarians and active surveillance is advised to explore its zoonotic potential