Generation of Random luxCDABE Transcriptional Fusions in the Genome of Salmonella enterica

Author(s):  
Juan J. de la Rosa-Altura ◽  
Barbora Benesova ◽  
Concepción Panadero-Medianero ◽  
Aida Amador-Álvarez ◽  
Julia Aguilera-Herce ◽  
...  
Microbiology ◽  
2009 ◽  
Vol 155 (8) ◽  
pp. 2490-2497 ◽  
Author(s):  
F. Gil ◽  
I. Hernández-Lucas ◽  
R. Polanco ◽  
N. Pacheco ◽  
B. Collao ◽  
...  

OmpW of Salmonella enterica serovar Typhimurium has been described as a minor porin involved in osmoregulation, and is also affected by environmental conditions. Biochemical and genetic evidence from our laboratory indicates that OmpW is involved in efflux of and resistance towards paraquat (PQ), and its expression has been shown to be activated in response to oxidative stress. In this study we have explored ompW expression in response to PQ. Primer extension and transcriptional fusions showed that its expression was induced in the presence of PQ. In silico analyses suggested a putative binding site for the SoxS transcriptional factor at the ompW regulatory region. Electrophoretic mobility shift assays (EMSAs) and footprinting experiments showed that SoxS binds at a region that starts close to −54 and ends at about −197 upstream of the transcription start site. Transcriptional fusions support the relevance of this region in ompW activation. The SoxS site is in the forward orientation and its location suggests that the ompW gene has a class I SoxS-dependent promoter.


2007 ◽  
Vol 190 (5) ◽  
pp. 1658-1670 ◽  
Author(s):  
I. Hernández-Lucas ◽  
A. L. Gallego-Hernández ◽  
S. Encarnación ◽  
M. Fernández-Mora ◽  
A. G. Martínez-Batallar ◽  
...  

ABSTRACT LeuO is a LysR-type transcriptional regulator that has been implicated in the bacterial stringent response and in the virulence of Salmonella. A genomic analysis with Salmonella enterica serovar Typhi revealed that LeuO is a positive regulator of OmpS1, OmpS2, AssT, and STY3070. In contrast, LeuO down-regulated the expression of OmpX, Tpx, and STY1978. Transcriptional fusions supported the positive and negative LeuO regulation. Expression of ompS1, assT, and STY3070 was induced in an hns mutant, consistent with the notion that H-NS represses these genes; transcriptional activity was lower for tpx and STY1978 in an hns background, suggesting that this global regulatory protein has a positive effect. In contrast, ompS2 and ompX expression appeared to be H-NS independent. LeuO specifically bound to the 5′ intergenic regions of ompS2, assT, STY3070, ompX, and tpx, while it was not observed to bind to the promoter region of STY1978, suggesting that LeuO regulates in direct and indirect ways. In this work, a novel set of genes belonging to the LeuO regulon are described; interestingly, these genes are involved in a variety of biological processes, suggesting that LeuO is a global regulator in Salmonella.


2001 ◽  
Vol 183 (19) ◽  
pp. 5733-5742 ◽  
Author(s):  
Bindhu Michael ◽  
Jenee N. Smith ◽  
Simon Swift ◽  
Fred Heffron ◽  
Brian M. M. Ahmer

ABSTRACT Proteins of the LuxR family detect the presence ofN-acylhomoserine lactones (AHLs) and regulate transcription accordingly. When AHLs are synthesized by the same species that detects them, the system allows a bacterium to measure the population density of its own species, a phenomenon known as quorum sensing. The sdiA genes of Escherichia coli and Salmonella enterica serovar Typhimurium are predicted to encode LuxR homologs. However, these species do not appear to synthesize AHLs or any other molecule detected by SdiA. It has previously been demonstrated that overexpression ofsdiA results in the activation of theftsQAZ locus in E. coli and four other loci in Salmonella serovar Typhimurium. Here we report that transcriptional fusions to these five loci fall into two classes. The first class requires overexpression of sdiA for activation. The second class responds to sdiA expressed from its natural position in the chromosome if the appropriate AHLs are added to the culture. The only member of the second class is a series of Prck-luxCDABE fusions in Salmonellaserovar Typhimurium. SdiA responds with highest sensitivity to AHLs that have a keto modification at the third carbon and an acyl chain length of 6 or 8 (half-maximal response between 1 and 5 nM). Growth ofSalmonella in proximity to species known to synthesize these AHLs results in sdiA-dependent activation of the Prck-luxCDABE fusions. SdiA appears to be the first AHL receptor discovered that detects signals emanating exclusively from other species.


Author(s):  
Dr. Manish Kulshrestha ◽  
Dr. Anjali Kulshrestha

INTRODUCTION: Enteric fever includes typhoid and paratyphoid fever. Peak incidence is seen in children 5–15 years of age; but in regions where the disease is highly endemic, as in India, children younger than 5 years of age may have the highest infection rates. There are about 22 million new typhoid cases occur each year. Young children in poor, resource limited areas, who make up the majority of the new cases and there is a mortality figures of 215,000 deaths annually. A sharp decline in the rates of complications and mortality due to typhoid fever is observed as a result of introduction of effective antibiotic therapy since 1950s. MDR-ST became endemic in many areas of Asia, including India soon after multidrug-resistant strains of Salmonella enterica serotype typhi (MDR-ST) that were resistant to all the three first-line drugs then in use, namely chloramphenicol, amoxycillin and co-trimoxazole emerged in early 1990s. MATERIAL AND METHODS: Only blood culture or bone marrow culture positive cases were included. The patients with culture isolated enteric fever were included in the study. Antimicrobial susceptibility testing was carried out by disk diffusion method using antibiotic discs. The analysis of the antimicrobial susceptibility was carried out as per CLSI interpretative guidelines. RESULTS: A total of 82 culture positive cases were included in the present study. 80 culture isolates were from blood culture and 2 from the bone marrow culture. Salmonella entericasubspecies enterica serovartyphi (S typhi) was isolated from 67 (81.70%) patients while Salmonella enterica subspecies entericaserovarparatyphi (S paratyphi A) was isolated from 13 (15.85%) cases and 2 (2.44%) were Salmonella enterica subspecies entericaserovarschottmuelleri (S paratyphi B). Of the 82 cases 65(79.3%) isolates were resistant to ciprofloxacin, 17 (20.7%) were resistant to nalidixic acid, one (1.2%) case each was resistant to Cefotaxime and ceftriaxone, 2 (2.4%) were resistant to chloramphenicol, 10 (12.2%) were resistant and to cotrimoxazole 3 (3.7%) were resistant. CONCLUSION: In a culture positive cases 65(79.3%) isolates were resistant to ciprofloxacin and 17 (20.7%) were resistant to nalidixic acid. Multidrug resistant isolates were 65(79.3%).


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