Generation of Left-handed Z-DNA in Solution and Visualization in Polytene Chromosomes by Immunofluorescence

Jovin, Thomas M.; Sande, J.H. van de; Zarling, David A.; Arndt‐Jovin, Donna J.; Eckstein, Fritz; Füldner, H.H.; Greider, Carol W.; Grieger, I.; Hamori, Eugene; Kalisch, Bernd W.; McIntosh, Lawrence P.; Robert‐Nicoud, Michel

doi:10.1101/sqb.1983.047.01.019

Cold Spring Harbor Symposia on Quantitative Biology

1983

DOI: 10.1101/sqb.1983.047.01.019

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Generation of Left-handed Z-DNA in Solution and Visualization in Polytene Chromosomes by Immunofluorescence

Thomas M. Jovin¹,

J.H. van de Sande²,

David A. Zarling³

et al.

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1983

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Cited by 73 publications

(43 citation statements)

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“…The assignments we derive independently for (dC-dG)3 and (dC-dG)4 duplexes in high salt agree with those poly(dG-dC) assignments (19), although our spectra are much better resolved as a result of the lower molecular weight of the oligonucleotide. This correspondence occurs despite problems in the former study due to: (i) assumptions about the similarity in response of the thioand oxo-phosphates to conformational change and (ii) problems associated with bulk magnetic susceptibility corrections necessary to relate chemical shifts measured in low and high salt with respect to an external standard.…”

supporting

confidence: 71%

“…Recently the 31P resonances of poly(dG-dC) (19) and poly-(dA-dT) (7) in high salt solutions were assigned by selective substitution or the phosphate group by phosphorothioate. The assignments we derive independently for (dC-dG)3 and (dC-dG)4 duplexes in high salt agree with those poly(dG-dC) assignments (19), although our spectra are much better resolved as a result of the lower molecular weight of the oligonucleotide.…”

mentioning

confidence: 99%

“…Also, 31P NMR spectra of the left handed Z DNA of polynucleotides show two characteristic signals with separation or ca. 0.5-1.5 ppm, which can be used to detect the presence of Z DNA in solution (3,6,8,9,(15)(16)(17)(18)(19)(20)(21)(22)(23). It is usual to assume that the downfield and upfield components correspond to predominantly gt and gg conformations, respectively, of the i and a torsion angles (24) in the 03'-P-05' linkages (6,(15)(16)(17)(18)(19)(20)(21)(22)(23) …”

mentioning

confidence: 99%

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³¹P-NMR analysis of the B to Z transition in double-stranded (dC-dG)₃, and (dC-dG)₄in high salt solution

Holak¹,

Borer²,

Levy³

et al. 1984

Nucl Acids Res

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ARITRACTIn 4M NaCl solutions (dC-dG) NMR spectroscopy gives evidence for sequence-dependent conformations or right handed deoxynucleotides (1-9). Assignments have recently been made for phosphodiester groups in the sugar-phosphate backbones ot snort oligonucleotides using 31P-lH decoupling experiments (10-12), two dimensional NMR 31P-lH correlation spectroscopy (13), and from 170 labeled experiments (14). Also, 31P NMR spectra of the left handed Z DNA of polynucleotides show two characteristic signals with separation or ca. 0.5-1.5 ppm, which can be used to detect the presence of Z DNA in solution (3,6,8,9,(15)(16)(17)(18)(19)(20)(21)(22)(23). It is usual to assume that the downfield and upfield components correspond to predominantly gt and gg conformations, respectively, of the i and a torsion angles (24) in the 03'-P-05' linkages (6,(15)(16)(17)(18)(19)(20)(21)(22)(23)

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supporting

confidence: 71%

mentioning

confidence: 99%

mentioning

confidence: 99%

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³¹P-NMR analysis of the B to Z transition in double-stranded (dC-dG)₃, and (dC-dG)₄in high salt solution

Holak¹,

Borer²,

Levy³

et al. 1984

Nucl Acids Res

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“…transcription (10) as well as recombination (32). In addition, since poly(dG-dT) forms Z-DNA under certain conditions (22), it has been reported to be involved in the definition of chromosomal domains through alterations in chromatin structure (14,23).Human alphoid DNA was chosen as a biologically important repeat family. It consists of related centromere sequences present at > 105 copies, accounting for several percent of the human genome (5, 37).…”

mentioning

confidence: 99%

Chromosome instability associated with human alphoid DNA transfected into the Chinese hamster genome.

Heartlein¹,

Knoll²,

Latt³

1988

Mol. Cell. Biol.

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Repetitive DNA sequences have been implicated in the mediation of DNA rearrangement in mammalian cells. We have tested this hypothesis by using a dihydrofolate reductase (DHFR) expression vector into which candidate sequences were inserted. DHFR-Chinese hamster ovary (CHO) cells were transfected with this vector, the amplification of which was then selected for by methotrexate (MTX) exposure. Cells transfected with the vector alone (and resistant to 0.02 or 1.0 ,uM MTX) or with a poly(dG-dT) insert (and resistant to 0.05 or 1.0 ,uM MTX) showed little change in chromosome aberrations or sister chromatid exchange frequencies. In contrast, transfection of DHFR-CHO cells with a vector containing either of two distinct 0.34-kilobase human alphoid DNA segments (and selection to 0.05 to 10.0 ,uM MTX) showed an approximately 50% increase in chromosome number and marked changes in chromosome structure, including one or two dicentric or ring forms per cell. The sister chromatid exchange frequency also increased, to more than double the frequency of that in cells transfected without insert or those containing poly(dG-dT). In situ hybridization of one 0.34-kilobase insert in some cells suggested clustering of homologous sequences in structurally abnormal recipient CHO cell chromosomes. The approach described provides an introduction to a unique means for a coordinate molecular and cytological study of dynamic changes in chromosome structure.Repetitive DNA in mammalian genomes represents a high proportion of total nuclear DNA (for reviews, see references 20 and 29). The DNA of chromosomal regions such as centromeres and telomeres, as well as that of constitutive heterochromatin, consists to a large degree of repetitive sequences which might somehow relate to functional characteristics, e.g., chromosome replication, pairing, and stability (2, 19). In addition, repeated DNA appears to be frequently associated with DNA reorganization (6,19,28,30), which in turn underlies some chromosomal rearrangements.Dihydrofolate reductase (DHFR)-based plasmid vectors (15-17), which can be transfected into DHFR-deficient cells and selectively amplified by methotrexate (MTX), provide one means of deliberately perturbing chromosomal DNA content and structural behavior. Previously it was reported that in DHFR-Chinese hamster ovary (DUKX) (34) cells transfected with a plasmid expressing DHFR, there was an increase in homogeneously staining regions and dicentric chromosomes at relatively high levels of MTX and of vector amplification (17). There was little associated change in sister chromatid exchange (SCE) frequency. This effect of the parent vector on chromosomes serves as a base line from which to observe the impact of different repeat sequence subunits which can be inserted into these plasmids and then coamplified with the dhfr gene. We have initiated a study of the effect of such DNA sequences on chromosome structure, as a function of their state of amplification. Specifically, it was possible to test for effects of two such sequences, the...

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“…6, 10-15, 29, 32, and reviewed in refs. 1,[3][4][5]. Assuming that the vector DNA remains in a right- The nine plasmids shown in Fig.…”

mentioning

confidence: 99%

Consecutive A X T pairs can adopt a left-handed DNA structure.

McLean¹,

Blaho²,

Kilpatrick³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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The capacity of six sequences with different numbers and orientations of APT pairs flanked by alternating C'G pairs to adopt left-handed structures was evaluated in recombinant plasmids. A series of synthetic oligodeoxynucleotides were cloned into the BamHI site of pRW790, a small plasmid (=%2 kilobases) prepared especially for conformational studies of this type. Supercoil relaxation studies by twodimensional gel electrophoresis on topoisomers of each plasmid revealed the energetics and structures of the left-handed helices. Also, the presence of supercoil-induced altered DNA conformations within the inserts of topoisomer populations of the plasmids was detected by reaction with S1 nuclease followed by restriction mapping of the cleavage sites. We conclude that consecutive TEA base pairs, whether alternating (TATA) or contiguous (TTTT), can adopt a left-handed conformation (presumably Z) when flanked by reasonably short runs of alternating (C-G). (n = 3-5).Thus, these results substantially broaden the range of DNA sequences that can adopt left-handed Z conformations.

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Generation of Left-handed Z-DNA in Solution and Visualization in Polytene Chromosomes by Immunofluorescence

Cited by 73 publications

References 0 publications

³¹P-NMR analysis of the B to Z transition in double-stranded (dC-dG)₃, and (dC-dG)₄in high salt solution

³¹P-NMR analysis of the B to Z transition in double-stranded (dC-dG)₃, and (dC-dG)₄in high salt solution

Chromosome instability associated with human alphoid DNA transfected into the Chinese hamster genome.

Consecutive A X T pairs can adopt a left-handed DNA structure.

Contact Info

Product

Resources

About

Generation of Left-handed Z-DNA in Solution and Visualization in Polytene Chromosomes by Immunofluorescence

Cited by 73 publications

References 0 publications

31P-NMR analysis of the B to Z transition in double-stranded (dC-dG)3, and (dC-dG)4in high salt solution

31P-NMR analysis of the B to Z transition in double-stranded (dC-dG)3, and (dC-dG)4in high salt solution

Chromosome instability associated with human alphoid DNA transfected into the Chinese hamster genome.

Consecutive A X T pairs can adopt a left-handed DNA structure.

Contact Info

Product

Resources

About

³¹P-NMR analysis of the B to Z transition in double-stranded (dC-dG)₃, and (dC-dG)₄in high salt solution

³¹P-NMR analysis of the B to Z transition in double-stranded (dC-dG)₃, and (dC-dG)₄in high salt solution