Guanine tetraplex topology of human telomere DNA is governed by the number of (TTAGGG) repeats

Secondary structures of the G-rich strand of human telomere DNA fragments G(3)(TTAG(3))(n), n = 1–16, have been studied by means of circular dichroism spectroscopy and PAGE, in solutions of physiological potassium cation concentrations. It has been found that folding of these fragments into tetraplexes as well as tetraplex thermostabilities and enthalpy values depend on the number of TTAG(3) repeats. The suggested topologies include, e.g. antiparallel and parallel bimolecular tetraplexes, an intramolecular antiparallel tetraplex, a tetraplex consisting of three parallel chains and one antiparallel chain, a poorly stable parallel intramolecular tetraplex, and both parallel and antiparallel tetramolecular tetraplexes. G(3)(TTAG(3))(3) folds into a single, stable and very compact intramolecular antiparallel tetraplex. With an increasing repeat number, the fragment tetraplexes surprisingly are ever less thermostable and their migration and enthalpy decrease indicate increasing irregularities or domain splitting in their arrangements. Reduced stability and different topology of lengthy telomeric tails could contribute to the stepwise telomere shortening process

Arrangements of human telomere DNA quadruplex in physiologically relevant K+ solutions

The arrangement of the human telomeric quadruplex in physiologically relevant conditions has not yet been unambiguously determined. Our spectroscopic results suggest that the core quadruplex sequence G3(TTAG3)3 forms an antiparallel quadruplex of the same basket type in solution containing either K+ or Na+ ions. Analogous sequences extended by flanking nucleotides form a mixture of the antiparallel and hybrid (3 + 1) quadruplexes in K+-containing solutions. We, however, show that long telomeric DNA behaves in the same way as the basic G3(TTAG3)3 motif. Both G3(TTAG3)3 and long telomeric DNA are also able to adopt the (3 + 1) quadruplex structure: Molecular crowding conditions, simulated here by ethanol, induced a slow transition of the K+-stabilized quadruplex into the hybrid quadruplex structure and then into a parallel quadruplex arrangement at increased temperatures. Most importantly, we demonstrate that the same transitions can be induced even in aqueous, K+-containing solution by increasing the DNA concentration. This is why distinct quadruplex structures were detected for AG3(TTAG3)3 by X-ray, nuclear magnetic resonance and circular dichrosim spectroscopy: Depending on DNA concentration, the human telomeric DNA can adopt the antiparallel quadruplex, the (3 + 1) structure, or the parallel quadruplex in physiologically relevant concentrations of K+ ions

Independent Regulatory Authorities in the Czech Republic from 2001 until the Present Day

This work is will deal with the amount of costs and personal provisions with a view to the performance of regulations of independent regulatory authorities in the Czech Republic from 2001 until the present day. Independent regulatory authorities are characteristic for their specific placement within the hierarchy of state administration, their independence from the political will, and they are financed from the state budget. This work aims to find out what is the proportion of costs related to the output of regulations of independent regulatory authorities to the overall costs linked to the output of state administration, and analyze the development of this indicator in the aforesaid period. In the same way, the development in the personnel area will be analysed. As the source for data analysis will be primarily used annual reports of selected regulatory authorities, The Czech Statistical Authority and Ministry of Finance. The amount of regulations by the state and reinforcing or streamlining of the functions of the state administrations -- these are the topics of constant scholarly discussions both in the economic and political fields. At present, The European Commission requires that the Czech Republic establish two new independent regulatory authorities in the fields of railway transport and water management. The reason to establish an independent regulatory authority shall be to make sure of equal conditions of liberalized market of railway transportation and functioning of independent regulatory authority in the water-management field, as stated in the conditions of European Commission for subsidizing in the period between 2014 and 2020. The theoretical part will define the terms of regulations and describe the concepts of economic theories related to the regulations controlled by the state. Furthermore, the theoretical part will focus on definitions of independent regulatory authority and will describe the historical development of independent regulatory authorities. The practical part will concern itself with the institutional and legal framework, competency and activities of independent regulatory authorities. Furthermore, their budget and personnel aspects will be analysed. The goal of the Thesis is to analyse the development of costs and personnel scope of independent regulatory authorities with a view to the overall costs linked to the state administration and to the overall personnel provisions of the state administration in the mentioned period

Independent Regulatory Authorities in the Czech Republic from 2001 until the Present Day

Diplomová práce se bude zabývat velikostí nákladů a personálním zabezpečením spojeným s výkonem regulace nezávislých regulačních úřadů v ČR od roku 2001 do současnosti. Nezávislé regulační úřady jsou charakterizovány specifickým zařazením v hierarchii státní správy, nezávislostí na politické vůli a jsou financované ze státního rozpočtu. Cílem práce je zjistit, jaký poměr tvoří náklady spojené s výkonem regulace nezávislých regulačních úřadů na celkových nákladech spojených s výkonem státní správy a analyzovat vývoj tohoto ukazatele v daném období. Stejně bude také analyzován vývoj v personální oblasti. Zdrojem pro analýzu dat budou zejména výroční zprávy vybraných regulačních úřadů, Český statistický úřad a Ministerstvo financí. Míra státní regulace a posilování či zeštíhlování funkcí státního aparátu jsou tématy, o kterých jsou neustále vedeny odborné diskuse na poli ekonomickém a politickém. Evropská komise aktuálně požaduje po ČR vytvoření dvou nových nezávislých regulačních úřadů v odvětví železniční dopravy a vodohospodářském. Důvodem zřízení nezávislého regulátora má být zajištění rovných podmínek liberalizovaného trhu železniční dopravy a fungováním nezávislého regulačního úřadu ve vodohospodářském odvětví podmiňuje Evropská komise čerpání dotací pro období 2014 až 2020. V teoretické části budou definovány pojmy regulace a budou zde popsány koncepty ekonomických teorií relevantních pro státem řízenou regulaci. Dále se teoretická část zaměří na definici nezávislý regulační úřad a nastíní historický vývoj nezávislých regulačních autorit. Praktická část přiblíží institucionálně právní rámec, kompetence a činnosti nezávislých regulačních úřadů. Dále bude analyzována jejich rozpočtová a personální stránka. Cílem práce je analýza vývoje nákladové a personální velikosti nezávislých regulačních úřadů na celkových nákladech státní správy a na celkovém personálním zabezpečení státní správy ve sledovaném období.This work is will deal with the amount of costs and personal provisions with a view to the performance of regulations of independent regulatory authorities in the Czech Republic from 2001 until the present day. Independent regulatory authorities are characteristic for their specific placement within the hierarchy of state administration, their independence from the political will, and they are financed from the state budget. This work aims to find out what is the proportion of costs related to the output of regulations of independent regulatory authorities to the overall costs linked to the output of state administration, and analyze the development of this indicator in the aforesaid period. In the same way, the development in the personnel area will be analysed. As the source for data analysis will be primarily used annual reports of selected regulatory authorities, The Czech Statistical Authority and Ministry of Finance. The amount of regulations by the state and reinforcing or streamlining of the functions of the state administrations -- these are the topics of constant scholarly discussions both in the economic and political fields. At present, The European Commission requires that the Czech Republic establish two new independent regulatory authorities in the fields of railway transport and water management. The reason to establish an independent regulatory authority shall be to make sure of equal conditions of liberalized market of railway transportation and functioning of independent regulatory authority in the water-management field, as stated in the conditions of European Commission for subsidizing in the period between 2014 and 2020. The theoretical part will define the terms of regulations and describe the concepts of economic theories related to the regulations controlled by the state. Furthermore, the theoretical part will focus on definitions of independent regulatory authority and will describe the historical development of independent regulatory authorities. The practical part will concern itself with the institutional and legal framework, competency and activities of independent regulatory authorities. Furthermore, their budget and personnel aspects will be analysed. The goal of the Thesis is to analyse the development of costs and personnel scope of independent regulatory authorities with a view to the overall costs linked to the state administration and to the overall personnel provisions of the state administration in the mentioned period

The guanine-rich fragile X chromosome repeats are reluctant to form tetraplexes

Using circular dichroism spectroscopy, UV absorption spectroscopy and polyacrylamide gel electrophoresis, we studied conformational properties of guanine-rich DNA strands of the fragile X chromosome repeats d(GGC)(n), d(GCG)(n) and d(CGG)(n), with n = 2, 4, 8 and 16. These strands are generally considered in the literature to form guanine tetraplexes responsible for the repeat expansion. However, we show in this paper that the repeats are reluctant to form tetraplexes. At physiological concentrations of either Na(+) or K(+) ions, the hexamers and dodecamers associate to form homoduplexes and the longer repeats generate homoduplexes and hairpins. The tetraplexes are rarely observed being relatively most stable with d(GGC)(n) and least stable with d(GCG)(n). The tetraplexes are exclusively formed in the presence of K(+) ions, at salt concentrations higher than physiological, more easily at higher than physiological temperatures, and they arise with extremely long kinetics (even days). Moreover, the capability to form tetraplexes sharply diminishes with the oligonucleotide length. These facts make the concept of the tetraplex appearance in this motif in vivo very improbable. Rather, a hairpin of the fragile X repeats, whose stability increases with the repeat length, is the probable structure responsible for the repeat expansion in genomes

Influence of additional bases and of a redox marker on the structure of the Thrombin Binding Aptamer

info:eu-repo/semantics/nonPublishe

Specific interaction between a redox marker and G quartets

info:eu-repo/semantics/nonPublishe