Left-handed Z-DNA in bands of acid-fixed polytene chromosomes

D. J. Arndt-Jovin, M. Robert-Nicoud, D. A. Zarling, Carol W Greider, E. Weimer, T. M. Jovin

Research output: Contribution to journalArticle

Abstract

Antibodies to DNA in the left-handed (Z) conformation bind to acid-fixed polytene chromosomes of both Chironomus thummi and Drosophila melanogaster, as shown by direct and indirect immunofluorescence. Comparison of the phase-contrast, immunofluorescence, and DNA staining patterns shows a predominant localization of the antibody to the regions of high contrast and DNA density, the bands. The immunofluorescence is completely abolished by competition with polynucleotides in the Z conformation but not by those in the B form. DNase but not RNase treatment eliminates the antibody staining. Actinomycin D inhibits binding, whereas mithramycin has no effect. The highly reproducible immunofluorescence patterns obtained with the anti-Z-DNA antibodies demonstrate variations in fluorescence intensity between particular bands, which can be quantitated by laser scanning and photon counting techniques. The telomeric regions and DNA strands associated with end-to-end chromosome linkage and ectopic pairing are exceptionally bright. At saturation, average values of 1 IgG molecule per 3,000 base pairs and 1 per 15,000 base pairs are found in the intensely and weakly staining regions, respectively. An alternative statement is that the left-handed Z-DNA conformation is present at a frequency of 0.02-0.1%. The measured differences reflect variations in the local density of Z-DNA sites and not in the affinity for the specific antibody, which appears to be relatively constant throughout the chromosomes (K(d) ≃ 10 nM). These observations taken together with results of biophysical studies on the properties of Z-DNA in solution suggest that regions of DNA in the left-handed conformation could be involved in higher-order structural organization of chromosomes and possibly in modulation of their functional state.

Original languageEnglish (US)
Pages (from-to)4344-4348
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume80
Issue number14 I
StatePublished - 1983
Externally publishedYes

Fingerprint

Z-Form DNA
Polytene Chromosomes
Acids
Fluorescent Antibody Technique
Chromosomes
Staining and Labeling
Base Pairing
Antibodies
DNA
Plicamycin
Chironomidae
Nucleic Acid Conformation
Polynucleotides
Direct Fluorescent Antibody Technique
Antibody Affinity
Deoxyribonucleases
Antinuclear Antibodies
Dactinomycin
Indirect Fluorescent Antibody Technique
Ribonucleases

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Arndt-Jovin, D. J., Robert-Nicoud, M., Zarling, D. A., Greider, C. W., Weimer, E., & Jovin, T. M. (1983). Left-handed Z-DNA in bands of acid-fixed polytene chromosomes. Proceedings of the National Academy of Sciences of the United States of America, 80(14 I), 4344-4348.

Left-handed Z-DNA in bands of acid-fixed polytene chromosomes. / Arndt-Jovin, D. J.; Robert-Nicoud, M.; Zarling, D. A.; Greider, Carol W; Weimer, E.; Jovin, T. M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 80, No. 14 I, 1983, p. 4344-4348.

Research output: Contribution to journalArticle

Arndt-Jovin, DJ, Robert-Nicoud, M, Zarling, DA, Greider, CW, Weimer, E & Jovin, TM 1983, 'Left-handed Z-DNA in bands of acid-fixed polytene chromosomes', Proceedings of the National Academy of Sciences of the United States of America, vol. 80, no. 14 I, pp. 4344-4348.
Arndt-Jovin, D. J. ; Robert-Nicoud, M. ; Zarling, D. A. ; Greider, Carol W ; Weimer, E. ; Jovin, T. M. / Left-handed Z-DNA in bands of acid-fixed polytene chromosomes. In: Proceedings of the National Academy of Sciences of the United States of America. 1983 ; Vol. 80, No. 14 I. pp. 4344-4348.
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