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B to Z transitions of the short DNA hairpins formed from the oligomer sequences: d[(CG)3X4(CG)3] (X = A, T, G, C). 总被引:1,自引:1,他引:0
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Circular Dichroism (CD) spectra were collected as a function of sodium perchlorate concentration [NaClO4] for the set of DNA hairpins formed from the oligomer sequences d[(CG)3X4(CG)3] where X = A, T, G or C. Over the range in salt concentration from 0 to 4.0 M NaClO4, the CD spectra invert in a manner characteristic of the B to Z transition. A factor analysis routine is described and employed to determine the least number of basis spectra required to fit the measured spectra of each hairpin over the entire salt range examined. In every case, linear combinations of only two sub-spectra fit the experimental spectra of the hairpins with greater than 98% accuracy, indicating the spectrally monitored structural transitions are two-state. From the relative weights of the individual sub-spectra, B-Z transition curves are constructed. The transitions are analyzed in terms of a simple two-state equilibrium model which yields an evaluation of the transition free-energy, delta GB-Z, as a function of NaClO4 concentration. At 1.0 M NaClO4 and 21 degrees C, delta GB-Z = 5.4, 4.9, 3.6 and 2.3 kcal/mole for the G4, T4, A4 and C4 loop hairpins, respectively. 相似文献
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Influence of a mechanical tension on the B-A and B-C conformational transitions in DNA fibres 总被引:1,自引:0,他引:1
In the present fibre X-ray study we attempt to quantify the effect of a mechanical tension on the conformations, and transitions between the structural forms of DNA. A simple experimental device has been realized in order to apply precise mechanical forces on DNA fibres during X-ray exposure. It is shown that, as the applied tension is increased, the B----A transition can be prevented as well as with a decrease of the sodium salt content. A kind of distorted B form is then observed the helical parameters of which change with the relative humidity. On the contrary, the mechanical tension does not prevent the B----C transition; it only slows down the form change and improves the X-ray patterns up to a relative humidity of 0%. 相似文献
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Slight changes in conditions influence the family of non-B-DNA conformations of the herpes simplex virus type 1 DR2 repeats 总被引:7,自引:0,他引:7
The segment inversion site of herpes simplex virus type 1 contains a series of tandem repeats with a purine bias on one strand and high G + C content (DR2 repeats) capable of adopting a non-B-DNA structure under a variety of conditions. Plasmids carrying eight contiguous copies of DR2 sequences undergo a series of supercoil-driven conformational transitions resulting in different extents of relaxation at pH 5.0. These transitions depend on the presence of an appropriate concentration of divalent cations (Mg2+ and Ca2+) which seem to interact specifically with the alternate structure(s). The transitions occurred at approximately the same superhelical density for all lengths of inserts studied. However, the onset of the transition can be shifted to lower negative superhelical densities by increasing NaCl concentrations. This leads to a reduction of the cooperativity of the transition, which takes place over a range of linking isomers under these conditions. Extrapolating from these results, we established physiological conditions where the alternate DNA structure is found at negative superhelical densities as low as -0.035. The existence of non-B-DNA conformations and/or the structural transitions of these sequences located in this region of intense biological activity implies their involvement in the life cycle of the virus. 相似文献
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pUC12-Wl is a new cloning vector for the study of torsion-induced structural transitions of insert DNA. It was derived from pUC12 by deleting three A + T-rich sequences which can undergo structural transitions when torsionally stressed. Transitions at these sites have low energy of activation and undefined structures. They complicate studies on transitions of DNA inserts by diverting torsional force and causing the vector to be undefined in helical and energetic terms. The new vector pUC12-Wl, from which these segments have been deleted, will facilitate studies of torsion-induced structural transitions of insert DNA. 相似文献
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The small subunit ribosomal RNA gene (srDNA) has been used extensively for phylogenetic analyses. One common assumption in these analyses is that substitution rates are biased toward transitions. We have developed a simple method for estimating relative rates of base change that does not assume rate constancy and takes into account base composition biases in different structures and taxa. We have applied this method to srDNA sequences from taxa with a noncontroversial phylogeny to measure relative rates of evolution in various structural regions of srRNA and relative rates of the different transitions and transversions. We find that: (1) the long single-stranded regions of the RNA molecule evolve slowest, (2) biases in base composition associated with structure and phylogenetic position exist, and (3) the srDNAs studied lack a consistent transition/transversion bias. We have made suggestions based on these findings for refinement of phylogenetic analyses using srDNA data. 相似文献
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G. Albiser M. Harmouchi S. Premilat 《Journal of biomolecular structure & dynamics》2013,31(2):359-366
Abstract In the present fibre X-ray study we attempt to quantify the effect of a mechanical tension on the conformations, and transitions between the structural forms of DNA A simple experimental device has been realized in order to apply precise mechanical forces on DNA fibres during X-ray exposure. It is shown that, as the applied tension is increased, the B→A transition can be prevented as well as with a decrease of the sodium salt content A kind of distorted B form is then observed the helical parameters of which change with the relative humidity. On the contrary, the mechanical tension does not prevent the B→C transition; it only slows down the form change and improves the X-ray patterns up to a relative humidity of 0%. 相似文献
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Protonated polypurine/polypyrimidine DNA tracts that appear to lack the single-stranded pyrimidine loop predicted by the "H" model 总被引:2,自引:0,他引:2
Three synthetic oligomers: 5'd(AG)8.dA.d(CT)(8)3'(A), 5'd(TC)7.d(TTA).d(GA)(8)3'(B) and d(GA)17(C) were cloned into the plasmid vector p915 in order to study the effects of sequence symmetry on pH-dependent structural transitions in polypurine/polypyrimidine DNA. When present in linear molecules all three sequences undergo transitions to protonated states. These are kinked to different degrees as determined by a non-denaturing gel mobility shift assay. Chemical probe analysis shows that the protonated states adopted by the linear forms of A and C exhibit certain features which have been regarded as indicating partially triple stranded "H" transition structures. The chemical reactivities of the transition structure adopted by linear molecule B and certain features of those exhibited by the transition structures of linear molecules A and C do not conform to the predictions of the "H" model. 相似文献
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The influence of tertiary structural restraints on conformational transitions in superhelical DNA.
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C J Benham 《Nucleic acids research》1987,15(23):9985-9995
This paper examines theoretically the effects that restraints on the tertiary structure of a superhelical DNA domain exert on the energetics of linking and the onset of conformational transitions. The most important tertiary constraint arises from the nucleosomal winding of genomic DNA in vivo. Conformational transitions are shown to occur at equilibrium at less extreme superhelicities in DNA whose tertiary structure is restrained than in unrestrained molecules where the residual linking difference alpha res (that part of the superhelical deformation which is not absorbed by transitions) may be freely partitioned between twisting and bending. In the extreme case of a rigidly held tertiary structure, this analysis predicts that the B-Z transition will occur at roughly half the superhelix density needed to drive the same transition in solution, other factors remaining fixed. This suggests that superhelical transitions may occur at more moderate superhelical deformations in vivo than in solution. The influence on transition behavior of the tertiary structural restraints imposed by gel conditions also are discussed. 相似文献
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The formation of melted regions from A + T-rich sequences and left-handed Z-DNA by alternating purine-pyrimidine sequences will both be facilitated by negative supercoiling, and thus if the sequences are present within the same plasmid molecule they will compete for the free energy of supercoiling. We have studied a series of plasmids that contain either (CG)8 or (TG)12 sequences in either G + C or A + T-rich contexts, by means of two-dimensional gel electrophoresis and chemical modification. We observe both B-Z and helix-coil transitions in all plasmids at elevated temperatures and low ionic strength. The plasmids fall into a number of different classes, in terms of the conformational behavior. As the superhelix density is increased, pCG8/vec ((CG)8 in G + C-rich context) undergoes an initial B-Z transition, followed by melting transitions in sequences remote from the (CG)8 sequence. The two transitions are coupled through the topology of the molecule but are otherwise independent. When the (CG)8 sequence was placed in an A + T-rich context (pCG8/col), the helix-coil transition was perturbed by the presence of the Z-DNA segment. Replacement of the (CG)8 tracts by (TG)12 sequences resulted in a further level of interaction between the transitions. Statistical mechanical modeling of the transitions suggested that at intermediate levels of negative supercoiling the Z-DNA formed by the (TG)12 sequence has a lowered probability due to the helix-coil transition in the A + T-rich sequences. These studies illustrate the complexities of competing conformational equilibria in supercoiled DNA molecules. 相似文献
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On the basis of published measurements of the melting transitions of synthetic polydeoxyribonucleotides with known sequences we have determined the parameters of the interplane (stacking) interactions of base pairs in DNA over the range of ionic strengths from 0.01 to 0.1 M Na+. We found that deviations of the stacking-interaction energy from the mean value of 7-8 kcal/mole were extremely small and did not exceed 0.2 kcal/mole. We report an analysis of the influence of the heterogeneity of the stacking interactions on the melting parameters of polynucleotides with random sequences (models of natural DNA's). Inclusion of this effect does not significantly distort the linear dependence of the melting temperature on the relative content of G-C pairs and insignificantly affects the width of the helix-coil transition in DNA under normal conditions. However it is the heterogeneity of the stacking interactions that plays the crucial role in the melting of DNA under conditions where the difference between the relative stabilities of the A-T and G-C pairs tends to zero, as in concentrated solutions of tetraethylammonium and tetramethylammonium salts. 相似文献
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Craig J. Benham 《Journal of molecular biology》1981,150(1):43-68
This paper presents a theoretical analysis of the conformation of a torsionally deformed segment of DNA containing two sites susceptible to stress-induced transitions in secondary structure. A mechanical analysis of the ensuing competitive behavior is developed and applied to several phenomena of possible biological relevance. First, a molecular lesion which disrupts base pairing without strand breakage (such as a pyrimidine dimer) is shown to provide an effective nucleation site for further stress-induced denaturation. A competition is established between strand separation at this lesion site and at the A + T-richest portion of the molecule. The relative importance of these two forms of melting is shown to depend upon the A + T-content of the sites involved, segment length, local environmental conditions and the magnitude of the imposed torsional deformation. A possible alternative mode of behavior of a stressed segment of DNA involves transitions from B-form to Z-form. The second application of this theory analyzes the interplay between B → Z transitions and local denaturation in torsionally stressed DNA. Finally, local melting is shown to be energetically preferred over transitions to A-form under physiologically reasonable conditions in vitro, due primarily to the greater degree of unwinding involved in melting.The mechanical theory presented here makes several simplifying assumptions regarding the nature of the transitions and the sequences involved. First, the theory is developed explicitly for the competition between two sites of possible transition, with no further consideration given to conformational degeneracy or sequence effects. These sites are regarded as being uniform in composition. A multistate, heteropolymeric statistical mechanical transition theory is required to account rigorously for degeneracy and the influence of base sequence. A preliminary formulation of such a theory is used to analyze the denaturation of a segment containing a site of disrupted base pairing. 相似文献
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Changes in chromatin structure during the aging of cell cultures as revealed by differential scanning calorimetry 总被引:1,自引:0,他引:1
Nuclei from cultured human cells were examined by differential scanning calorimetry. Their melting profiles revealed four structural transitions at 60, 76, 88, and 105 degrees C (transitions I-IV, respectively). In immortalized (i.e., tumor) cell cultures and in normal cell cultures of low passage number, melting profiles were dominated by the 105 degrees C transition (transition IV), but in vitro aging of normal and Werner syndrome cells was associated with a marked decrease in transition IV followed by an increase in transition III at the expense of transition IV. At intermediate times in the aging process, much DNA melted at a temperature range (95-102 degrees C) intermediate between transitions III and IV, and this is consistent with the notion that aging of cell cultures is accompanied by an increase in single-strand character of the DNA. Calorimetric changes were observed in the melting profile of nuclei from UV-irradiated tumor cells that resembled the age-induced intermediate melting of chromatin. It is suggested that aging is accompanied by an increase in single-stranded character of the DNA in chromatin, which lowers its melting temperature, followed by strand breaks in the DNA that destroy its supercoiling potential. 相似文献
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The appearance of the slow mode, revealed by dynamic light scattering (DLS) measurements in Micrococcus luteus DNA with high GC content, and the effect of guanine sequences on changes of DNA physical state and conformational transitions
were investigated. We used two different spectroscopic approaches: DLS, to evidence the relatively slowly diffusing particles
arising at high salt concentration, ascribable to the formation of large unspecific molecular aggregates, and circular dichroism
spectroscopy, to identify these entities. Our results bring us to conclude that a peculiar, unconventional, structural transition,
due to the presence of long guanine stretches, in a well-defined experimental condition, can occur. We comment on the biological
implications to detect, by spectroscopic measurements, such an unusual structure involved in the stability, protection and
replication maintenance along the human telomeric G-rich strand. 相似文献
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Substitution-rate variation among sites and differences in the
probabilities of change among the four nucleotides are conflated in DNA
sequence comparisons. When variation in rate exists among sites but is
ignored, biases in the rates of change among nucleotides are
underestimated. This paper provides a quantification of this effect when
the observed proportions of transitions, P, and transversions, Q, between
two sequences are used to estimate transition bias. The utility of P/Q as
an estimator is examined both with and without rate variation among sites.
A gamma-distributed-rates model is used to illustrate the effect that
variation among sites has on estimates of transition bias, but it is argued
that the basic results should hold for any pattern of rate variation. Naive
estimates of the extent of transition bias, those that ignore rate
variation when it is present, can seriously underestimate its true value.
The extent of this underestimation increases with the amount of rate
variation among sites. An example using human mitochondrial DNA shows that
a simple comparison of the proportions of transitions and transversions in
recently diverged sequences underestimates the level of transition bias by
approximately 15%. This does not depend on the use of P/Q to estimate
transition bias; maximum-likelihood methods give similar results.
相似文献
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The structural transitions of (Lysx, Leuy)n-DNA and (Lysx)n(Leuy)m-DNA complexes have been studied by thermal denaturation utilizing simultaneous absorption and circular dichroism (CD) measurements [R. Mandel and G.D. Fasman (1974), Biochem. Biophys. Res. Commun. 59, 672]. These complexes are used as models for nucleohistones. At amino acid/nucleotide ratios r less than 1, the copolymers bind to DNA in a ratio of one amino acid residue per nucleotide, and such binding stabilizes the DNA double helix against thermal denaturation relative to the unbound regions. The leucine residues in the copolymers stabilize the bound portion of the complex against thermal denaturation but to a lesser degree than does poly(L-lysine). This study confirms the hypothesis that absorption melting profiles reflect only the change in secondary structure (helix-coil transition) of DNA. It was found that, in the absence of a higher ordered structure (condensed), the CD melting profile also reflects this same conformational transition, and the melting temperatures, Tm, in CD are equal to those in absorption. However, when a higher ordered structure (tertiary) exists in the complex, then the CD melting profile will be dominated by the structural transitions related to the melting of the higher ordered asymmetric structure in the condensed state, followed by the melting of the secondary structure. Under such circumstances, the Tm obtained from absorption may be slightly different from that of the CD, since only the secondary structural changes are being reflected in absorption. The relevance of these studies to the structure of chromatin is discussed. 相似文献