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Transcription activation and DNA binding by the MelR proteinHoward, Victoria Jayne January 2001 (has links)
No description available.
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Bioinformatics and Biological Databases: 1) Sigma-54 Promoter Database – A Database of Sigma-54 Promoters Covering a Wide Range of Bacterial Genomes 2) ClusterMine360 – A Database of PKS/NRPS BiosynthesisConway, Kyle 14 January 2013 (has links)
The Sigma-54 Promoter Database contains computationally predicted sigma-54 promoters from
over 60 prokaryotic species. Organisms from all major phyla were analysed and results were
made available online at http://www.sigma54.ca. This database is particularly unique due to its
inclusion of intragenic regions, grouping of data by COG and COG category, and the ability to
summarize results either by phylum or database-wide.
ClusterMine360 (http://www.clustermine360.ca/) is a database of microbial polyketide and nonribosomal peptide gene clusters. It takes advantage of crowd-sourcing by allowing members of the community to make contributions while automation is used to help achieve high data
consistency and quality. The database currently has over 200 gene clusters from over 185
compound families. It also features a unique sequence repository containing over 10,000
PKS/NRPS domains. The sequences are filterable and downloadable as individual or multiple
sequence FASTA files. This database will be a useful resource for members of the PKS/NRPS
research community enabling them to keep up with the growing number of sequenced gene
clusters and rapidly mine these clusters for functional information.
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Bioinformatics and Biological Databases: 1) Sigma-54 Promoter Database – A Database of Sigma-54 Promoters Covering a Wide Range of Bacterial Genomes 2) ClusterMine360 – A Database of PKS/NRPS BiosynthesisConway, Kyle 14 January 2013 (has links)
The Sigma-54 Promoter Database contains computationally predicted sigma-54 promoters from
over 60 prokaryotic species. Organisms from all major phyla were analysed and results were
made available online at http://www.sigma54.ca. This database is particularly unique due to its
inclusion of intragenic regions, grouping of data by COG and COG category, and the ability to
summarize results either by phylum or database-wide.
ClusterMine360 (http://www.clustermine360.ca/) is a database of microbial polyketide and nonribosomal peptide gene clusters. It takes advantage of crowd-sourcing by allowing members of the community to make contributions while automation is used to help achieve high data
consistency and quality. The database currently has over 200 gene clusters from over 185
compound families. It also features a unique sequence repository containing over 10,000
PKS/NRPS domains. The sequences are filterable and downloadable as individual or multiple
sequence FASTA files. This database will be a useful resource for members of the PKS/NRPS
research community enabling them to keep up with the growing number of sequenced gene
clusters and rapidly mine these clusters for functional information.
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Bioinformatics and Biological Databases: 1) Sigma-54 Promoter Database – A Database of Sigma-54 Promoters Covering a Wide Range of Bacterial Genomes 2) ClusterMine360 – A Database of PKS/NRPS BiosynthesisConway, Kyle January 2013 (has links)
The Sigma-54 Promoter Database contains computationally predicted sigma-54 promoters from
over 60 prokaryotic species. Organisms from all major phyla were analysed and results were
made available online at http://www.sigma54.ca. This database is particularly unique due to its
inclusion of intragenic regions, grouping of data by COG and COG category, and the ability to
summarize results either by phylum or database-wide.
ClusterMine360 (http://www.clustermine360.ca/) is a database of microbial polyketide and nonribosomal peptide gene clusters. It takes advantage of crowd-sourcing by allowing members of the community to make contributions while automation is used to help achieve high data
consistency and quality. The database currently has over 200 gene clusters from over 185
compound families. It also features a unique sequence repository containing over 10,000
PKS/NRPS domains. The sequences are filterable and downloadable as individual or multiple
sequence FASTA files. This database will be a useful resource for members of the PKS/NRPS
research community enabling them to keep up with the growing number of sequenced gene
clusters and rapidly mine these clusters for functional information.
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Expanded Functionality of the Bacterial Global Regulator LrpHart, Benjamin Randall 26 August 2010 (has links)
No description available.
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Inhibiteurs de la transcription bactérienne : étude du mécanisme d'action de la lipiarmycine et dépendance au facteur de transcription σ / Inhibitors of bacterial transcription : study of the mechanism of action of lipiarmycin and dependence on transcription factor σTupin, Audrey 19 November 2010 (has links)
Le nombre croissant de bactéries résistantes aux antibiotiques et le problème des cellules persistantes rend urgent le développement de nouveaux antibiotiques et la compréhension de leur mécanisme d'action. L'ARN polymérase est l'enzyme centrale de la transcription et est une cible intéressante pour les antibiotiques. Dans cette étude, nous nous sommes particulièrement intéressés à un inhibiteur de l'ARN polymérase : la lipiarmycine. Il s'agit d'un inhibiteur de la transcription macrocyclique qui inhibe les bactéries à Gram + et qui est en essai clinique de phase III pour le traitement des infections liées à Clostridium difficile. L'objectif de ce travail a été de déterminer le mécanisme d'action de la lipiarmycine ainsi que le mécanisme de résistance à la molécule. Pour cela, nous avons dans un premier temps précisé et modélisé son site de liaison sur l'ARN polymérase. Puis, dans un deuxième temps, nous avons utilisé des approches génétiques et biochimiques afin de déterminer son mécanisme et l'effet de certaines mutations sur la transcription. Ces travaux ont mis à jour un nouveau mécanisme d'inhibition de la transcription. / The growing number of antibiotic-resistant bacteria added to the problem caused by persistent cells stress the need for developing new antibiotics and for understanding their mechanism of action. RNA polymerase is the main enzyme of the transcription process and is an interesting target for antibiotics. In this study we focus on a particular inhibitor of RNA polymerase : lipiarmycin. It is a macrocyclic inhibitor of transcription inhibiting Gram + bacteria that is developed in phase III clinical trials for treatment of Clostridium difficile infections. The objective of this work was to determine the mechanism of action of lipiarmycin and the mechanism confering resistance against the molecule. We first define more precisely its binding site on RNA polymerase and then used genetic and biochemical approaches to determine its mechanism of action and the effect of some specific mutations on transcription. Our experiments reveal a new mechanism of t ranscription inhibitor action.
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Elucidating the Role of MsRbpA in Rifampicin Tolerance and Transcription Regulation of Mycobacterium SmegmatisVerma, Amit Kumar January 2013 (has links) (PDF)
RNA polymerase binding protein A (RbpA) was first discovered as a RNA polymerase binding protein from Streptomyces. coelicolor. It was shown to cause rifampicin tolerance to RNA polymerase in vitro and leads to basal level of rifampicin resistance in vivo. This protein is exclusively present in the actinobacteria family with the nearest neighbour in mycobacteria. When null mutant of RbpA in S. coelicolor were transformed with the rbpA gene from Mycobacterium tuberculosis the resistance level of rifampicin increased from 0.75 µgml-1 to 2 µg ml-1 suggesting analogous role of MtbRbpA (RbpA from M. tuberculosis). MsRbpA, RbpA from Mycobacterium smegmatis was found to interact with the β-subunit of RNAP and its binding location on M. smegmatis RNAP was shown to be 18 Å from the (i+1) site. MsRbpA was also shown to rescue the inhibitory effect of rifampicin in vitro. Furthermore, overexpression of MsRbpA in wild type M. smegmatis resulted in the increase in the MIC of rifampicin to 85 µg ml-1 from 20 µg ml-1, which is the MIC of rifampicin for the wild type M. smegmatis. On the other hand, MsRbpA was unable to augment transcription in the presence of rifampicin when the reaction was catalysed by rifampicin resistant RNAP. Recent reports have shown that MtbRbpA enhances the affinity σA to core RNAP thereby activates transcription. The N and C-termini of MtbRbpA interact with σA while the C-terminal region of MtbRbpA is required for the oligomerisation of MtbRbpA. However M. tuberculosis and S. coleicolor are part of same family actinobacteria, RbpA is essential for the former while it is dispensable in the later case.This work focuses on characterisation of rifampicin resistant RNAP from M. smegmatis and elaborates on the roles played by MsRbpA. These include its effect on transcription activation, transcription rescue, its role in RNAP promoter closed and open complex formation, characterisation of its site of interaction with RNAP and σA, finding critical functional residues and establishing the essentiality of MsRbpA in M. smegmatis.
Chapter 1 deals with the literature survey on structure of bacterial RNAP, promoters, sigma factors, RNAP inhibitors, transcriptional activators with the emphasis on the Mycobacteria.
Chapter 2 summarises the identification of the mutations in rpoB gene from the rifampicin resistant (RifR) mutant strains of M. smegmatis, purification of RNAP from these strain, determining IC50 values of these RifR RNAP for rifampicin, finding kinetic parameters for the interaction of RifR RNAP with 3-formyl rifampicin and evaluating their interaction with MsRbpA.
Chapter 3 describes the function of MsRbpA in transcription initiation, particularly its role in RNAP-promoter closed and open complex formation. Furthermore, this chapter throws light on the role of MsRbpA in transcription activation vis a vis its effects on transcription rescue from the inhibitory effect of rifampicin.
Chapter 4 elucidates the function of a segment of MsRbpA from Arg58 to Lys 73 in activation of transcription activity, transcription rescue from the inhibitory effect of rifampicin and its interaction with σA and core RNAP. Furthermore, the alanine scanning of the region and subsequent in vitro transcription studies revealed four important residues required for MsRbpA functions.
Chapter 5 describes the generation of conditional knock down strain of MsRbpA in M. smegmatis and establishing its essentiality.
Chapter 6 summarizes the work documented in the thesis.
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Insights into Occurrence and Divergence of Intrinsic Terminators and Studies on Rho-Dependent Termination in Mycobacterium TuberculosisMitra, Anirban January 2013 (has links) (PDF)
Two mechanisms, intrinsic and factor-dependent, have evolved for accomplishing the termination of transcription in eubacteria. In this thesis, the first chapter is an introduction to the topic that presents what is known about the mechanisms of termination. The properties of the primary and secondary ‘players’- intrinsic terminators, Rho protein, rho-dependent terminators, RNA polymerse and Nus factors - are presented and the known mechanisms by which termination functions are discussed. In Chapter 2, a detailed analysis of intrinsic terminators – their differential distribution, similarity and divergence - has been penned. The database, compiled using the program GeSTer (Genome Scanner for Terminators), comprises ~2000 sequences and is one of the largest of its kind. Furthermore, analyzing the data from over 700 bacteria reveals how different species have fine-tuned intrinsic terminators to suit their cellular needs. Non-canonical intrinsic terminators emerge to be a significant fraction of the observed structures. The conserved structural features of identified intrinsic terminators are discussed and the relationship between the two modes of termination is assessed. Chapter 3 deals with the importance of transcription termination in regulating horizontally acquired DNA. The results show that genomic islands are scarce in intrinsic terminators and thus constitute most likely sites for Rho-dependent termination. Plausible reasons for why such a scenario has evolved are discussed and a generally applicable model is presented. Chapters 4 and 5 focus on Rho protein from Mycobacterium tuberculosis. In silico identification of M. tuberculosisgenes that rely on MtbRho-dependent termination is followed by experimental validation. The data show that Rho-dependent termination is the predominant mechanism in this species.MtbRho is a majorly expressed protein that governs termination of protein-coding and non-protein coding genes. Further, MtbRho can productively interact with RNA that has considerable secondary structure. Such interactions cause conformational changes in the enzyme. Given that MtbRho has to function with a GC-rich transcriptome, the altered properties could have evolved for optimal function.
Taken together, the thesis extends our current understanding of both modes of termination. The importance of non-canonical intrinsic terminators in mycobacteria and other organisms is discussed. The unusual function of Rho and its predominant role in mycobacteria is elucidated. Finally, the inter-relationship between the two modes of termination is also discussed.
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