Detection of long-range sequence order in Drosophila melanogaster satellite DNA IV by a photochemical crosslinking reaction and denaturation microscopy

Che Kun James Shen, John E. Hearst

研究成果: 雜誌貢獻文章同行評審

21 引文 斯高帕斯(Scopus)

摘要

Satellite DNA IV of Drosophila melanogaster has been isolated in an actinomycin/CsCl gradient and allowed to react with tritium-labeled 4,5′,8-trimethylpsoralen (trioxsalen) in the presence of long wavelength ultraviolet light. Saturation experiments showed a limiting covalent binding of approximately one trioxsalen molecule per three base-pairs for the photochemical reaction. The buoyant density change of satellite IV (ρ = 1·705 g/cm3) after photoreaction was found to be -0·182 (g/cm3)/(trioxsalen/base-pair). To visualize directly the interstrand crosslinks, the photochemically reacted satellite DNA was denatured and spread for electron microscopy. From 85% to 90% of the molecules examined have an appearance of long stretches of tandem loops with few observable double-stranded regions. The histogram of loop sizes in these denatured molecules shows a regular distribution of crosslinkable sites along satellite DNA IV with an interval of approximately 250 base-pairs, suggesting that a long-range periodicity of sequences exists in this satellite DNA. After extensive crosslinking, the observed loops still have sizes that are multiples of 250 base-pairs. These results are discussed in terms of the sequence of satellite DNA IV, the characteristics of the photochemical reaction, and theories of evolution of satellite DNA sequences.

原文英語
頁(從 - 到)495-507
頁數13
期刊Journal of Molecular Biology
112
發行號3
DOIs
出版狀態已發佈 - 五月 25 1977
對外發佈Yes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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