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Evolution of RAYT/REP system in Stenotrophomonas
Staněk, Tadeáš ; Nunvář, Jaroslav (advisor) ; Zahradník, Jiří (referee)
REP (repetitive extragenic palindromes) are short DNA motifs found in the intergenic regions of many bacteria, typically in the hundreds of copies per genome. Unable to mobilize on their own, REP are propagated by RAYT (REP-associated tyrosine transposase). Stenotrophomonas are G- bacteria with a broad ecological niche, belonging to the Gammaproteobacteria. RAYT/REP systems are widespread in them, with over 132,000 REP found in 102 strains. Several classes of REP coexist in the genomes, the presence of associated RAYT is essential for high REP abundance. REP are localised in "REP loci", a series of intergenic regions with high evolutionary dynamics - the number, sequence, and orientation of REP vary at a given locus. In addition to genomic islands, REP are absent from the replication termination site, a region showing non-standard evolutionary dynamics (variable number and order of genes), probably related to chromosome segregation. Stenotrophomonas are insertion sequences (IS) rich. 35 unique IS were identified and their insertion target was verified. Ten IS insert into REP, and four other target YPAL (Yersinia palindromic elements), DNA motifs so far known only in the genus Yersinia. A total of 14 out of 35 IS in Stenotrophomonas target intergenic motifs, while their copy number accounts for 75%...
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Y1 and Y2 transposases, mechanisms of transposition, biological function.
Zahradník, Jiří ; Lichá, Irena (advisor) ; Schierová, Michaela (referee)
Transposases are enzymes that catalyse cleavage, transmission and re-inserting of mobile genetic element into the DNA. Tyrosine transposase take between these enzymes completely independend status. Their uniqueness is determined by their structure and different mechanism of the transposition reaction, in which the covalent phosphotyrosine intermediate plays major role. Mandatory presence of the catalytic tyrosine gives name to these enzymes and it enables their further classification into a group that carries only a single catalytic tyrosine - Y1 transposases and a group carrying two tyrosines - Y2 transposases. This thesis summarizes the current knowledge about tyrosine transposases. It covers their occurrence, structure, reaction mechanism and biological function. The reaction mechanism of the most studied Y1 transposase, associated with IS608 element, is described in detail. The work also focuses on other members of the tyrosin transposases family which carry the characteristic HUH motive. These include transposases associated with the insertion sequence of IS200/IS605 family (Y1), transposases associated with REP elements (so called RAYT proteins), transposases associated with IS91 family (Y2), transposases of ISCRs family (Y1) and unusual eukaryotic transposases of the Helitron family (Y2)....
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Bacterial REP elements: origins, variability and application.
Nunvář, Jaroslav ; Lichá, Irena (advisor) ; Pačes, Jan (referee) ; Melter, Oto (referee)
4 ABSTRACT (English) This thesis is based on three published research papers studying bacterial REP (repetitive extragenic palindrome) elements. REP elements are one of the best-characterized groups of bacterial DNA repeats, distributed mostly in gammaproteobacteria, including enterobacteria. They are present in noncoding parts of host genomes, usually occurring in hundreds of copies. REPs are typically aggregated in higher order repeats. In the Gram-negative model Escherichia coli, interactions of several proteins important for cell's physiology with REPs were described, indicating significant role for these elements for host cells. The first work (Nunvar et al. 2010) presents the discovery of a protein class, related to IS200/IS605 transposases. These proteins, termed RAYTs (REP-associated tyrosine transposases), contain characteristic motifs in their amino acid sequences, which are absent in canonical IS200/IS605 transposases. Another attribute of RAYTs is the arrangement of their encoding genes. These are single copy genes, always flanked at both termini by at least two REPs in inverted orientation. Based on the similarity between the REP-rayt-REP unit and insertion sequences of the IS200/IS605 family, between RAYTs and tyrosine transposases and between REPs and subterminal sequences of the IS200/IS605...
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Bacterial REP elements: origins, variability and application.
Nunvář, Jaroslav
4 ABSTRACT (English) This thesis is based on three published research papers studying bacterial REP (repetitive extragenic palindrome) elements. REP elements are one of the best-characterized groups of bacterial DNA repeats, distributed mostly in gammaproteobacteria, including enterobacteria. They are present in noncoding parts of host genomes, usually occurring in hundreds of copies. REPs are typically aggregated in higher order repeats. In the Gram-negative model Escherichia coli, interactions of several proteins important for cell's physiology with REPs were described, indicating significant role for these elements for host cells. The first work (Nunvar et al. 2010) presents the discovery of a protein class, related to IS200/IS605 transposases. These proteins, termed RAYTs (REP-associated tyrosine transposases), contain characteristic motifs in their amino acid sequences, which are absent in canonical IS200/IS605 transposases. Another attribute of RAYTs is the arrangement of their encoding genes. These are single copy genes, always flanked at both termini by at least two REPs in inverted orientation. Based on the similarity between the REP-rayt-REP unit and insertion sequences of the IS200/IS605 family, between RAYTs and tyrosine transposases and between REPs and subterminal sequences of the IS200/IS605...
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Bacterial REP elements: origins, variability and application.
Nunvář, Jaroslav
4 ABSTRACT (English) This thesis is based on three published research papers studying bacterial REP (repetitive extragenic palindrome) elements. REP elements are one of the best-characterized groups of bacterial DNA repeats, distributed mostly in gammaproteobacteria, including enterobacteria. They are present in noncoding parts of host genomes, usually occurring in hundreds of copies. REPs are typically aggregated in higher order repeats. In the Gram-negative model Escherichia coli, interactions of several proteins important for cell's physiology with REPs were described, indicating significant role for these elements for host cells. The first work (Nunvar et al. 2010) presents the discovery of a protein class, related to IS200/IS605 transposases. These proteins, termed RAYTs (REP-associated tyrosine transposases), contain characteristic motifs in their amino acid sequences, which are absent in canonical IS200/IS605 transposases. Another attribute of RAYTs is the arrangement of their encoding genes. These are single copy genes, always flanked at both termini by at least two REPs in inverted orientation. Based on the similarity between the REP-rayt-REP unit and insertion sequences of the IS200/IS605 family, between RAYTs and tyrosine transposases and between REPs and subterminal sequences of the IS200/IS605...
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Bacterial REP elements: origins, variability and application.
Nunvář, Jaroslav ; Lichá, Irena (advisor) ; Pačes, Jan (referee) ; Melter, Oto (referee)
4 ABSTRACT (English) This thesis is based on three published research papers studying bacterial REP (repetitive extragenic palindrome) elements. REP elements are one of the best-characterized groups of bacterial DNA repeats, distributed mostly in gammaproteobacteria, including enterobacteria. They are present in noncoding parts of host genomes, usually occurring in hundreds of copies. REPs are typically aggregated in higher order repeats. In the Gram-negative model Escherichia coli, interactions of several proteins important for cell's physiology with REPs were described, indicating significant role for these elements for host cells. The first work (Nunvar et al. 2010) presents the discovery of a protein class, related to IS200/IS605 transposases. These proteins, termed RAYTs (REP-associated tyrosine transposases), contain characteristic motifs in their amino acid sequences, which are absent in canonical IS200/IS605 transposases. Another attribute of RAYTs is the arrangement of their encoding genes. These are single copy genes, always flanked at both termini by at least two REPs in inverted orientation. Based on the similarity between the REP-rayt-REP unit and insertion sequences of the IS200/IS605 family, between RAYTs and tyrosine transposases and between REPs and subterminal sequences of the IS200/IS605...
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Y1 and Y2 transposases, mechanisms of transposition, biological function.
Zahradník, Jiří ; Lichá, Irena (advisor) ; Schierová, Michaela (referee)
Transposases are enzymes that catalyse cleavage, transmission and re-inserting of mobile genetic element into the DNA. Tyrosine transposase take between these enzymes completely independend status. Their uniqueness is determined by their structure and different mechanism of the transposition reaction, in which the covalent phosphotyrosine intermediate plays major role. Mandatory presence of the catalytic tyrosine gives name to these enzymes and it enables their further classification into a group that carries only a single catalytic tyrosine - Y1 transposases and a group carrying two tyrosines - Y2 transposases. This thesis summarizes the current knowledge about tyrosine transposases. It covers their occurrence, structure, reaction mechanism and biological function. The reaction mechanism of the most studied Y1 transposase, associated with IS608 element, is described in detail. The work also focuses on other members of the tyrosin transposases family which carry the characteristic HUH motive. These include transposases associated with the insertion sequence of IS200/IS605 family (Y1), transposases associated with REP elements (so called RAYT proteins), transposases associated with IS91 family (Y2), transposases of ISCRs family (Y1) and unusual eukaryotic transposases of the Helitron family (Y2)....
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