scholarly journals Identification of a core sequence for the binding of BosR to the rpoS promoter region in Borrelia burgdorferi

Microbiology ◽  
2015 ◽  
Vol 161 (4) ◽  
pp. 931-931 ◽  
Author(s):  
Zhiming Ouyang ◽  
Ranjit K. Deka ◽  
Michael V. Norgard ◽  
Jianli Zhou ◽  
Chad A. Brautigam
Keyword(s):  
Borrelia Burgdorferi ◽  
Promoter Region ◽  
Core Sequence

scholarly journals Identification of a core sequence for the binding of BosR to the rpoS promoter region in Borrelia burgdorferi

Microbiology ◽  
2014 ◽  
Vol 160 (5) ◽  
pp. 851-862 ◽  
Author(s):  
Zhiming Ouyang ◽  
Jianli Zhou ◽  
Chad A. Brautigam ◽  
Ranjit Deka ◽  
Michael V. Norgard
Keyword(s):  
Borrelia Burgdorferi ◽  
Transcriptional Activation ◽  
Promoter Region ◽  
Regulatory Function ◽  
Adaptive Responses ◽  
Mobility Shift ◽  
Core Sequence ◽  
Dyad Symmetry ◽  
Electrophoretic Mobility Shift Assays ◽  
Binding Sequence

The alternative sigma factor RpoS in Borrelia burgdorferi plays a central role in modulating host adaptive responses when spirochaetes cycle between ticks and mammals. The transcriptional activation of σ54-dependent rpoS requires a Fur homologue designated BosR. Previously, BosR was shown to directly activate rpoS transcription by binding to the rpoS promoter. However, many other DNA binding features of BosR have remained obscure. In particular, the precise DNA sequence targeted by BosR has not yet been completely elucidated. The prediction of a putative Per box within the rpoS promoter region has further confounded the identification of the BosR binding sequence. Herein, by using electrophoretic mobility shift assays, we demonstrate that the putative Per box predicted in the rpoS promoter region is not involved in the binding of BosR. Rather, a 13 bp palindromic sequence (ATTTAANTTAAAT) with dyad symmetry, which we denote as the ‘BosR box’, functions as the core sequence recognized by BosR in the rpoS promoter region of Borrelia burgdorferi. Similar to a Fur box and a Per box, the BosR box probably comprises a 6–1–6 inverted repeat composed of two hexamers (ATTTAA) in a head-to-tail orientation. Selected mutations in the BosR box prevented recombinant BosR from binding to rpoS. In addition, we found that sequences neighbouring the BosR box also are required for the formation of BosR–DNA complexes. Identification of the BosR box advances our understanding of how BosR recognizes its DNA target(s), and provides new insight into the mechanistic details behind the unique regulatory function of BosR.

scholarly journals Effects of Environmental Changes on Expression of the Oligopeptide Permease (opp) Genes of Borrelia burgdorferi

2002 ◽  
Vol 184 (22) ◽  
pp. 6198-6206 ◽  
Author(s):  
Xing-Guo Wang ◽  
Bo Lin ◽  
J. Michael Kidder ◽  
Samuel Telford ◽  
Linden T. Hu
Keyword(s):  
Borrelia Burgdorferi ◽  
Promoter Region ◽  
Nitrogen Availability ◽  
Environmental Changes ◽  
Substrate Binding ◽  
Environmental Response ◽  
Promoter Regions ◽  
Promoter Sequences ◽  
Oligopeptide Permease

ABSTRACT We analyzed expression of a putative oligopeptide permease (Opp) of Borrelia burgdorferi. Unlike the opp operons of other bacteria for which there is a single substrate binding protein, B. burgdorferi codes for three substrate binding proteins (OppA-I to -III) in its opp operon and an additional two homologs on plasmids (OppA-IV and -V). Instead of a single promoter region regulating transcription of the entire operon, as seen in other bacterial opp operons, it appears that among oppA-I, -II, and -III, as well as oppA-IV and -V, each has a potential upstream promoter region. We tested the function of these putative promoter sequences by fusion to a promoterless β-galactosidase reporter gene in pCB182. Each of the promoter regions was found to be active. The level of activity in the reporter constructs closely paralleled the level of expression of each gene in in vitro-grown B. burgdorferi. Changes in carbon and nitrogen availability differentially affected individual promoters, but no changes in promoter activity were seen when Escherichia coli bacteria (with the promoter constructs) were grown in various concentrations of phosphate and leucine and changes in pH. Expression of specific oppA genes with B. burgdorferi varied significantly between its mouse and fed and unfed tick hosts. Differences in regulation of opp gene expression suggest a potential role in environmental response by the organism.

scholarly journals DNA-Binding Proteins Possibly Involved in Regulation of the Post-Logarithmic-Phase Expression of Lipoprotein P35 in Borrelia burgdorferi

2000 ◽  
Vol 182 (2) ◽  
pp. 522-525 ◽  
Author(s):  
Karl J. Indest ◽  
Mario T. Philipp
Keyword(s):  
Dna Binding ◽  
Borrelia Burgdorferi ◽  
Stationary Phase ◽  
Growth Phase ◽  
Promoter Region ◽  
Protein Complexes ◽  
Phase Cell ◽  
Logarithmic Phase ◽  
Specific Complex

ABSTRACT Previously, we have shown that the transcription ofp35, a lipoprotein gene of Borrelia burgdorferi, is upregulated or initiated during the post-logarithmic bacterial growth phase in vitro. To identify potential regulatory factors, we examined the formation of DNA-protein complexes by electromobility shift assay, using a 157-bp DNA fragment that spans the p35 promoter region and cell-free extracts of spirochetes harvested from both logarithmic and stationary growth phases. The binding properties of the extracts with the promoter region of the flaB gene, a constitutively expressed, growth-phase-independent gene, were also compared. The results from these experiments demonstrate that B. burgdorferistationary-phase cell-free extracts have a growth-phase-dependent DNA binding protein that interacts specifically with the p35promoter region. We show, in addition, that a segment from the 157-bpp35 promoter region which contains both a T-rich stretch and an inverted repeat is able to compete off the stationary-phase-specific complex when the segment is present in molar excess.

New ultrastructural findings about Borrelia burgdorferi by cryofixation, negative staining, thin sectioning and scanning techniques

1990 ◽  
Vol 48 (3) ◽  
pp. 582-583
Author(s):  
S. F. Hayes ◽  
M. D. Corwin ◽  
T. G. Schwan ◽  
D. W. Dorward ◽  
W. Burgdorfer
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Structural Characterization ◽  
Negative Staining ◽  
Low Speed ◽  
Thin Sectioning ◽  
Ultrastructural Findings

Characterization of Borrelia burgdorferi strains by means of negative staining EM has become an integral part of many studies related to the biology of the Lyme disease organism. However, relying solely upon negative staining to compare new isolates with prototype B31 or other borreliae is often unsatisfactory. To obtain more satisfactory results, we have relied upon a correlative approach encompassing a variety EM techniques, i.e., scanning for topographical features and cryotomy, negative staining and thin sectioning to provide a more complete structural characterization of B. burgdorferi.For characterization, isolates of B. burgdorferi were cultured in BSK II media from which they were removed by low speed centrifugation. The sedimented borrelia were carefully resuspended in stabilizing buffer so as to preserve their features for scanning and negative staining. Alternatively, others were prepared for conventional thin sectioning and for cryotomy using modified procedures. For thin sectioning, the fixative described by Ito, et al.

968: Gelsolin Gene Silencing Involving Unusual Chromatin Structures and a Novel Stem Loop Structure of the Promoter Region in Human Bladder Cancer

2004 ◽  
Vol 171 (4S) ◽  
pp. 256-257
Author(s):  
Kazunori Haga ◽  
Ataru Sazawa ◽  
Toru Harabayashi ◽  
Nobuo Shinohara ◽  
Minoru Nomoto ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Gene Silencing ◽  
Promoter Region ◽  
Loop Structure ◽  
Human Bladder Cancer ◽  
Human Bladder ◽  
Stem Loop ◽  
Stem Loop Structure

Variation in the Promoter Region of the β Fibrinogen Gene Is Associated with Plasma Fibrinogen Levels in Smokers and Non-Smokers

1991 ◽  
Vol 65 (05) ◽  
pp. 487-490 ◽  
Author(s):  
A E Thomas ◽  
F R Green ◽  
C H Kelleher ◽  
H C Wilkes ◽  
P J Brennan ◽  
...  
Keyword(s):  
Heart Disease ◽  
Promoter Region ◽  
Fragment Length Polymorphism ◽  
Fibrinogen Level ◽  
Plasma Fibrinogen ◽  
Plasma Fibrinogen Level ◽  
Healthy Men ◽  
History Of ◽  
General Practices ◽  
Restriction Fragment Length

SummaryWe investigated the association between fibrinogen levels and a HaeIII restriction fragment length polymorphism located at −453 bp from the start of transcription of the β fibrinogen gene. 292 healthy men aged 45 to 69 years, recruited from general practices throughout Britain, were studied. None had a history of ischaemic heart disease. 41.1% (120) were smokers and fibrinogen levels were higher in this group. The frequency of the noncutting allele (designated H2) was 0.19 and was the same in smokers and non-smokers. The H2 allele was associated with elevated levels of fibrinogen in both smokers and non-smokers and the effect of genotype was similar in both groups. After smoking, HaeIII genotype was the strongest predictor of fibrinogen levels and explained 3.1% of the variance in fibrinogen levels. These results confirm earlier studies that variation at the fibrinogen locus contributes to the between-individual differences in plasma fibrinogen level.

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