A5026838257- Mycobacterium research and diagnosis
- Tuberculosis Research and Epidemiology
- RNA and protein synthesis mechanisms
- Bacteriophages and microbial interactions
- Bacterial Genetics and Biotechnology
- Antibiotic Resistance in Bacteria
- CRISPR and Genetic Engineering
- Genomics and Phylogenetic Studies
- Chromosomal and Genetic Variations
- RNA modifications and cancer
- Advanced biosensing and bioanalysis techniques
- Microbial Community Ecology and Physiology
- DNA Repair Mechanisms
- DNA and Nucleic Acid Chemistry
- Diagnosis and treatment of tuberculosis
- Escherichia coli research studies
- Bacterial biofilms and quorum sensing
- Infectious Diseases and Tuberculosis
- Parasitic Infections and Diagnostics
- RNA regulation and disease
- Microbial Fuel Cells and Bioremediation
- Ubiquitin and proteasome pathways
- Genome Rearrangement Algorithms
- DNA and Biological Computing
- Graphite, nuclear technology, radiation studies
University at Albany, State University of New York
2013-2023
Wadsworth Center
2014-2023
New York State Department of Health
2014-2023
Albany State University
2013-2016
Western University
2005
Yale University
1984-1995
University of Edinburgh
1987
CITATION: Bitter, W., et al. 2009. Systematic genetic nomenclature for type VII secretion systems. PLoS Pathogens, 5(10): 1-6, doi: 10.1371/journal.ppat.1000507.
RNA-seq technologies have provided significant insight into the transcription networks of mycobacteria. However, such studies provide no definitive information on translational landscape. Here, we use a combination high-throughput transcriptome and proteome-profiling approaches to more rigorously understand protein expression in two mycobacterial species. ribosome profiling Mycobacterium smegmatis, start site (TSS) mapping N-terminal peptide mass spectrometry tuberculosis, complementary,...
Most bacterial ORFs are identified by automated prediction algorithms. However, these algorithms often fail to identify lacking canonical features such as a length of >50 codons or the presence an upstream Shine-Dalgarno sequence. Here, we use ribosome profiling approaches actively translated in Mycobacterium tuberculosis . have not been previously described, indicating that M. transcriptome is pervasively translated. The newly described predominantly short, with many encoding proteins...
Conjugal DNA transfer in Mycobacterium smegmatis occurs by a mechanism distinct from plasmid-mediated transfer. Previously, we had shown that the secretory apparatus, ESX-1, negatively regulated donor strain; ESX-1 mutants are hyper-conjugative. Here, describe genome-wide transposon mutagenesis screen to isolate recipient mutants. Surprisingly, find majority of insertions map within esx-1 locus, which encodes apparatus. Thus, contrast its role function, is essential for function;...
Conjugal DNA transfer occurs by an atypical mechanism in Mycobacterium smegmatis . The system is chromosomally encoded and requires recipient recombination functions for both chromosome plasmid transfer. Cis-acting sequences have been identified that confer mobility on nontransferable plasmids, but these are larger different properties to canonical oriT sites found bacterial plasmids. To identify trans-acting factors required mediating transfer, a library of transposon insertion mutants was...
Horizontal gene transfer (HGT) in bacteria generates variation and drives evolution, conjugation is considered a major contributor as it can mediate of large segments DNA between strains species. We previously described novel form chromosomal mycobacteria that does not conform to classic oriT-based models, whose potential evolutionary significance has been evaluated. Here, we determined the genome sequences 22 F1-generation transconjugants, providing first genome-wide view conjugal HGT at...
Significance Mycobacteria as well other bacteria remodel their ribosomes in response to zinc depletion by replacing zinc-binding ribosomal proteins with zinc-free paralogues, releasing for metabolic processes. In this study, we show that the remodeled ribosome acquires a structurally stable but functionally inactive and aminoglycoside-resistant state zinc-starved Mycobacterium smegmatis . Conversely, M. cells are growth arrested zinc-rich conditions have unstable reduced survival. We further...
Conseting mycobacteria Mycobacteria encompass several slow-growing pathogens, including organisms that cause leprosy and tuberculosis. use a component of their ESX (or Type VII) secretion system for distinctive form genetic exchange called distributive conjugal DNA transfer. Gray et al. investigated quicker-growing model species, Mycobacterium smegmatis. They found the secretory apparatus, ESX-4, is essential transfer into recipient but not required donor cells to pass along DNA. The ESX-1...
Summary The role of host factors in regulating bacterial transposition has never been comprehensively addressed, despite the potential consequences transposition. Here, we describe a screen for that influence IS 903 , and effect these mutations on two additional transposons, Tn 10 552 . Over 20 000 independent insertion mutants were screened strains Escherichia coli ; from isolated over 100 altered These included increased or decreased extent also timing during colony growth. large number...
The insertion sequence IS903 has perfect, 18-base-pair terminal repeats that are the presumed binding sites of its transposase. We have isolated mutations throughout this inverted repeat and analyzed their effect on transposition. show every position in (with possible exception 4) is important for efficient Furthermore, various substitutions at a single can wide range effects. Analysis these hierarchical effects suggests transposase contacts minor groove region from 13 to 16 but makes major...
IS6110 is an insertion element found exclusively within the members of Mycobacterium tuberculosis complex (MTBC), and because this exclusivity, it has become important diagnostic tool in identification MTBC species. The restriction to hypothesized arise from inability these bacteria exchange DNA. We have identified IS6110-related a strain smegmatis. presence indicates that lateral gene transfer occurred among mycobacterial species, suggesting pool larger than previously suspected.
Conjugal transfer of chromosomal DNA between strains Mycobacterium smegmatis occurs by a novel mechanism. In transposon mutagenesis screen, three transfer-defective insertions were mapped to the lsr2 gene donor strain mc(2)155. Because encodes nonspecific DNA-binding protein, mutations give rise variety phenotypes, including an inability form biofilms. this study, we show that efficient M. in mixed biofilm and process requires expression but not recipient strain. Testing cells from different...
The genus Mycobacterium includes the major human pathogens tuberculosis and leprae . development of rational drug treatments for diseases caused by these other mycobacteria requires establishment basic molecular techniques to determine genetic basis pathogenesis resistance. To date, ability manipulate move DNA between mycobacterial strains has relied on processes transformation transduction. Here, we describe a naturally occurring conjugation system present in smegmatis , which anticipate...
An unusual subset of DNA-binding proteins, termed cis-acting has been shown to act preferentially at their site synthesis; the transposases several bacterial insertion sequences (ISs) fall into this class. The transposase IS903 exhibits a strong preference for action in cis: complementation defective transposons trans occurs less than 1%. Furthermore, transposition mediated by acting cis is extremely sensitive distance between 3' end gene and nearest transposon inverted repeat; we find that...
Essential replication (rep) genes of the broad host range plasmid RSF1010 have been cloned onto controlled expression vectors and their protein products visualized, after induction, by NaDodSO4/polyacrylamide gel electrophoresis whole cell lysates. During this induction a coresident replicon, pKT210, was analyzed quantitative DNA X hybridization. The initiation pKT210 stimulated 6-fold simultaneous overproduction RepA RepC proteins compared to cells in which only overproduced. An enhanced...
Summary Most mycobacterial species spontaneously form biofilms, inducing unique growth physiologies and reducing drug sensitivity. Biofilm progresses through three genetically programmed stages: substratum attachment, intercellular aggregation architecture maturation. Growth of Mycobacterium smegmatis biofilms requires multiple factors including a chaperonin (GroEL1) nucleoid‐associated protein (Lsr2), although how their activities are linked remains unclear. Here it is shown that Lsr2...
Functional characterization of bacterial proteins lags far behind the identification new protein families. This is especially true for species that are more difficult to grow and genetically manipulate than model systems such as
Although prokaryotic organisms lack traditional organelles, they must still organize cellular structures in space and time, challenges that different species solve differently. To systematically define the subcellular architecture of mycobacteria, we perform high-throughput imaging a library fluorescently tagged proteins expressed Mycobacterium smegmatis develop customized computational pipeline, MOMIA GEMATRIA, to analyze these data. Our results establish spatial organization network over...
ABSTRACT Transposon mutagenesis in bacteria generally requires efficient delivery of a transposon suicide vector to allow the selection relatively infrequent transposition events. We have developed an IS 903 -based system for diverse gram-negative that is not limited by transfer efficiency. The transposon, 903φkan , carries cryptic kan gene, which can be expressed only after successful transposition. This allows stable introduction into host. Generation insertion mutants then frequency was...