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  • 1
    Language: English
    Source: National Library of Australia (Trove)〈img src=https://exlibris-pub.s3.amazonaws.com/Trove_reload.gif style="vertical-align:middle;margin-left:7px"〉
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  • 2
    In: Molecular Microbiology, August 2013, Vol.89(4), pp.715-731
    Description: –– and – are the major chaperone machineries in bacteria. In many species, and are encoded in bicistronic operons. Quantitative proteomics revealed that and amounts in dominate over and respectively. An imperfect transcriptional terminator in the intergenic region of is known to result in higher transcript levels of the first gene. Here, we examined the operon and asked how the second gene in a heat shock operon can be preferentially expressed and found that an structure in the 5′untranslated region of is responsible. The secondary structure masks the hine–algarno () sequence and start codon and thereby modulates translation of . Reporter gene assays combined with structure probing and toeprinting analysis revealed a dynamic temperature‐sensitive structure. Following an increase in temperature, only the second of two hairpins melts and partially liberates the sequence, thus facilitating translation. Translation of is not temperature‐regulated leading to an excess of the chaperonin in the cell at low temperature. Discussion in a broader context shows how structured segments can differentially control expression of temperature‐affected operons in various ways.
    Keywords: Translation (Genetics) -- Analysis ; Messenger Rna -- Analysis ; Genes -- Analysis ; Salmonella -- Analysis;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 3
    In: Molecular Microbiology, August 2014, Vol.93(3), pp.439-452
    Description: In contrast to numerous enzymes involved in c‐di‐ synthesis and degradation in enterobacteria, only a handful of c‐di‐ receptors/effectors have been identified. In search of new c‐di‐ receptors, we screened the   overexpression gene library using the ifferential adial apillary ction of igand ssay () with fluorescently and radioisotope‐labelled c‐di‐. We uncovered three new candidate c‐di‐ receptors in and characterized one of them, . The gene is encoded in cellulose synthase operons in representatives of ammaproteobacteria and etaproteobacteria. The purified proteins from , and bind c‐di‐ via the domain of unknown function, 2819, which is hereby designated , ‐site ike domain. The motif of the domain is required for c‐di‐ binding, similar to the c‐di‐‐binding ‐site of the diguanylate cyclase domain. Thus, is the second protein domain, after , dedicated to c‐di‐‐binding. We show that in  , is not essential for cellulose synthesis but is required for maximal cellulose production, and that c‐di‐ binding is critical for function. It appears that cellulose production in enterobacteria is controlled by a two‐tiered c‐di‐‐dependent system involving and the domain containing glycosyltransferase .
    Keywords: Proteins – Chemical Properties ; Protein Binding – Chemical Properties ; Escherichia Coli – Chemical Properties ; Cellulose – Chemical Properties;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 4
    In: European Integration Studies, 01/12/2012, Vol.0(5)
    Description: Elaboration and use of development policy monitoring instruments promotes applicable and effective policy implementation and decision making, which is based on regular and systematic verification of resources, actions and results. On different policy development levels exists varied approach of defining specific aims and use of policy monitoring measures. Since activation of development monitoring issues in Latvia, some institutions have gained experience in elaboration of monitoring systems among them Riga region and its local municipalities practise various approaches using policy monitoring instruments. Specific study analyzes theoretical aspects of different development monitoring approaches and factors that impact its quality. Monitoring performance evaluation is based mostly on territory development comparative analysis using the methods of statistical analysis by assessing territory development indicators. Practical examples explain experiences of development tendencies and policy evaluation as well as impact on further decision making and policy implementation. Analyze of development planning system laws and regulations explain legal basis impact on processes in the field of development policy elaboration, implementation and monitoring. Development monitoring and its role and effectiveness evaluation, and further monitoring system optimization analysis is an objective of this article. On different policy development levels exists varied approach on elaboration, implementation and monitoring of specific policy. This study analyzes the matter contextually on state, regional and local levels. Observations indicate link between development priorities and budget planning is a principally essential precondition for implementation of development policies. Different development monitoring approaches are determined by aim of the specific policy. Development evaluation mostly express as monitoring of general development processes, connected to specific aims or policies. Overall approach of development policy monitoring mostly is oriented on evaluation of planning documents. Riga region development monitoring practice is considered as successful, till now and within nearest future region's development monitoring realization based on statistics, thematic studies, efficiency analysis and functional monitoring is expected. Further research would be focused on new approaches on elaborating development monitoring instruments for suitability of use for specific aims on different policy development levels. Regional development monitoring system improvements would be connected with elaboration of operational indicators structure and the creation of flexible thematic indicators system. Permanently on the agenda of Riga region are monitoring themes such as public sector efficiency, the society's economic activity, social activity, population needs and movement. Specific thematic research is needed for solving challenges of environmental quality, human settlements and infrastructure planning, public transport, school network planning and other issues.
    Keywords: Economics;
    ISSN: 1822-8402
    E-ISSN: 23358831
    Source: CrossRef
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  • 5
    Language: English
    In: Journal of Molecular Biology, 14 September 2018, Vol.430(18), pp.3170-3189
    Description: Many bacteria secrete cellulose, which forms the structural basis for bacterial multicellular aggregates, termed biofilms. The cellulose synthase complex of Salmonella typhimurium consists of the catalytic subunits BcsA and BcsB and several auxiliary subunits that are encoded by two divergently transcribed operons, bcsRQABZC and bcsEFG. Expression of the bcsEFG operon is required for full-scale cellulose production, but the functions of its products are not fully understood. This work aimed to characterize the BcsG subunit of the cellulose synthase, which consists of an N-terminal transmembrane fragment and a C-terminal domain in the periplasm. Deletion of the bcsG gene substantially decreased the total amount of BcsA and cellulose production. BcsA levels were partially restored by the expression of the transmembrane segment, whereas restoration of cellulose production required the presence of the C-terminal periplasmic domain and its characteristic metal-binding residues. The high-resolution crystal structure of the periplasmic domain characterized BcsG as a member of the alkaline phosphatase/sulfatase superfamily of metalloenzymes, containing a conserved Zn2+-binding site. Sequence and structural comparisons showed that BcsG belongs to a specific family within alkaline phosphatase-like enzymes, which includes bacterial Zn2+-dependent lipopolysaccharide phosphoethanolamine transferases such as MCR-1 (colistin resistance protein), EptA, and EptC and the Mn2+-dependent lipoteichoic acid synthase (phosphoglycerol transferase) LtaS. These enzymes use the phospholipids phosphatidylethanolamine and phosphatidylglycerol, respectively, as substrates. These data are consistent with the recently discovered phosphoethanolamine modification of cellulose by BcsG and show that its membrane-bound and periplasmic parts play distinct roles in the assembly of the functional cellulose synthase and cellulose production. Unlabelled Image •BcsG subunit of cellulose synthase is required for full-scale cellulose production.•BcsG affects cellulose production via at least two distinct molecular mechanisms.•Transmembrane part of BcsG is required for proper production of the BcsA subunit.•The periplasmic domain of BcsG has the alkaline phosphatase superfamily structure.•Crystal structure of the BcsG periplasmic domain shows a single active-site Zn ion.
    Keywords: Alkaline Phosphatase Superfamily ; Biofilm Formation ; Cellulose Biosynthesis ; Extracellular Matrix ; Virulence ; C-Di-Gmp ; TM ; Alkp ; Pe ; PG
    ISSN: 0022-2836
    E-ISSN: 10898638
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  • 6
    Language: English
    In: Methods in molecular biology (Clifton, N.J.), 2017, Vol.1657, pp.225-241
    Description: The Congo Red (CR) assay is a standard biofilm test assessing the colony morphology of bacteria growing on agar plates supplemented with the diazo dye Congo Red. Biofilm forming Salmonella enterica serovar Typhimurium and Escherichia coli produce a red, dry, and rough (rdar) morphotype on CR-plates. The phenotype is characterized by staining of the extracellular matrix components curli (brown color) and cellulose (pink color) by CR. This method allows semiquantitative determination of the expression level of the individual matrix components and dissection of the regulatory networks controlling their production in response to c-di-GMP levels. Here, we describe the CR-assay and its variations and discuss the effect of deletion or overexpression of c-di-GMP turnover proteins on colony morphology.
    Keywords: Biofilm ; C-Di-Gmp ; Calcofluor ; Congo Red ; Rdar Morphotype ; Rugosity ; Biofilms ; Congo Red ; Escherichia Coli -- Growth & Development ; Salmonella Typhimurium -- Growth & Development
    E-ISSN: 1940-6029
    Source: MEDLINE/PubMed (U.S. National Library of Medicine)
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  • 7
    Language: English
    In: BMC Microbiology, 2017, Vol. 17
    Description: Background: The secondary messenger cyclic di-GMP promotes biofilm formation by up regulating the expression of csgD, encoding the major regulator of rdar biofilm formation in Salmonella typhimurium. The GGDEF/EAL domain proteins regulate the c-di-GMP turnover. There are twenty-two GGDEF/EAL domain proteins in the genome of S. typhimurium. In this study, we dissect the role of individual GGDEF/EAL proteins for csgD expression and rdar biofilm development. Results: Among twelve GGDEF domains, two proteins upregulate and among fifteen EAL domains, four proteins down regulate csgD expression. We identified two additional GGDEF proteins required to promote optimal csgD expression. With the exception of the EAL domain of STM1703, solely, diguanylate cyclase and phosphodiesterase activities are required to regulate csgD mediated rdar biofilm formation. Identification of corresponding phosphodiesterases and diguanylate cyclases interacting in the csgD regulatory network indicates various levels of regulation by c-di-GMP. The phosphodiesterase STM1703 represses transcription of csgD via a distinct promoter upstream region. Conclusion: The enzymatic activity and the protein scaffold of GGDEF/EAL domain proteins regulate csgD expression. Thereby, c-di-GMP adjusts csgD expression at multiple levels presumably using a multitude of input signals.
    Keywords: C-Di-Gmp ; Csgd ; Ggdef/Eal Domain Proteins ; Rdar Morphotype ; Biofilm Formation ; Salmonella Typhimurium ; Medical And Health Sciences ; Basic Medicine ; Microbiology In The Medical Area ; Medicin Och Hälsovetenskap ; Medicinska Och Farmaceutiska Grundvetenskaper ; Mikrobiologi Inom Det Medicinska Området
    ISSN: 1471-2180
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  • 8
    Language: English
    In: Journal of biotechnology, 2012, Vol.160(1), pp.55-63
    Description: Free-living bacteria constantly monitor their ambient temperature. Drastic deviations elicit immediate protective responses known as cold shock or heat shock response. Many mammalian pathogens use temperature surveillance systems to recognize the successful invasion of a host by its body temperature, usually 37°C. Translation of temperature-responsive genes can be modulated by RNA thermometers (RNATs). RNATs form complex structures primarily in the 5′-untranslated region of their transcripts. Most RNATs block the ribosome binding site at low temperatures. Translation is induced at increasing temperature by melting of the RNA structure. The analysis of such temperature-dependent RNA elements calls for adequate test systems that function in the appropriate temperature range. Here, we summarize previously established reporter gene systems based on the classical β-galactosidase LacZ, the heat-stable β-galactosidase BgaB and the green fluorescent protein GFP. We validate these systems by testing known RNATs and describe the construction and application of an optimized bgaB system. Finally, two novel RNA thermometer candidates from Escherichia coli and Salmonella will be presented. ; p. 55-63.
    Keywords: Mammals ; Green Fluorescent Protein ; Monitoring ; Binding Sites ; Reporter Genes ; Body Temperature ; Thermometers ; Pathogens ; Ribosomes ; Translation (Genetics) ; Biotechnology ; Beta-Galactosidase ; Rna ; Bacteria ; Escherichia Coli ; Cold Stress ; Ambient Temperature ; Gene Expression ; Heat Stability ; Salmonella ; Heat Shock Response ; Melting Point
    ISSN: 0168-1656
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  • 9
    Language: English
    In: 2017 14th Workshop on Positioning, Navigation and Communications (WPNC), October 2017, pp.1-6
    Description: Device-free localization (DFL) systems detect and track persons without devices that participate in the localization process. A person moving within a target area affects the electromagnetic field that is measured by received signal strength (RSS) values. Consequently for DFL systems modeling of RSS is important and still an open issue. In this paper, we develop a simple model for prediction of RSS values in a setup with transmitter and receiver devices, a person and multipath propagation. We design and implement the model as a superposition of both, knife-edge diffraction to account for the change made by the person, and, propagation effects such as multipath propagation that result in reflection and path loss including the antenna characteristics. We evaluate our model in comparison with real measurements in various setups with and without multipath propagation. We achieve an accuracy that is close to our hardware limitations, which is the resolution of the measured RSS values of the receiver.
    Keywords: Diffraction ; Receivers ; Transmitters ; Predictive Models ; Antennas ; Conferences ; Navigation ; Device-Free Localization Model ; Knife-Edge Diffraction ; Passive Object Tracking ; Propagation & Antennas ; Engineering
    Source: IEEE Conference Publications
    Source: IEEE Xplore
    Source: IEEE Journals & Magazines 
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  • 10
    Language: English
    In: Journal of Biotechnology, 31 July 2012, Vol.160(1-2), pp.55-63
    Description: Highlights► Several reporter gene systems can be used to analyze RNA thermometers in vivo. ► Both inducible as well as constitutively regulated systems are applicable. ► GFP and the β-galactosidases LacZ and BgaB (thermostable) are suitable reporters. ► Besides moderate heat shock, LacZ is optimal for analyzing cold shock regulation. ► Novel RNA thermometer candidates were found upstream of the htrA gene from Salmonella and Escherichia coli. Free-living bacteria constantly monitor their ambient temperature. Drastic deviations elicit immediate protective responses known as cold shock or heat shock response. Many mammalian pathogens use temperature surveillance systems to recognize the successful invasion of a host by its body temperature, usually 37°C. Translation of temperature-responsive genes can be modulated by RNA thermometers (RNATs). RNATs form complex structures primarily in the 5′-untranslated region of their transcripts. Most RNATs block the ribosome binding site at low temperatures. Translation is induced at increasing temperature by melting of the RNA structure. The analysis of such temperature-dependent RNA elements calls for adequate test systems that function in the appropriate temperature range. Here, we summarize previously established reporter gene systems based on the classical β-galactosidase LacZ, the heat-stable β-galactosidase BgaB and the green fluorescent protein GFP. We validate these systems by testing known RNATs and describe the construction and application of an optimized bgaB system. Finally, two novel RNA thermometer candidates from Escherichia coli and Salmonella will be presented.
    Keywords: Bp(S) ; Csp ; Fouru ; Hsp ; Nt ; Rbs ; Rnat ; Rose ; SD ; Tss ; Utr ; Cold Shock ; Heat Shock ; RNA Thermometer ; Translational Regulation ; Htra ; Protease
    ISSN: 0168-1656
    Source: ScienceDirect (Elsevier B.V.)
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