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First published online 13 July 2004
doi: 10.1242/jcs.01225

Journal of Cell Science 117, 3797-3805 (2004)
Published by The Company of Biologists 2004
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Research Article

Visualization of unconstrained negative supercoils of DNA on polytene chromosomes of Drosophila

Kuniharu Matsumoto1 and Susumu Hirose1,2,*

1 Department of Developmental Genetics, National Institute of Genetics, SOKENDAI, 1111 Yata, Mishima, Shizuokaken 411-8540, Japan
2 Department of Genetics, SOKENDAI, 1111 Yata, Mishima, Shizuokaken 411-8540, Japan

* Author for correspondence (e-mail: shirose{at}lab.nig.ac.jp)

Accepted 17 March 2004

Bulk DNA within the eukaryotic genome is torsionarily relaxed. However, unconstrained negative supercoils of DNA have been detected in few local domains of the genome through preferential binding of psoralen. To make a genome-wide survey for such domains, we introduced biotinylated psoralen into Drosophila salivary glands and visualized it on polytene chromosomes with fluorescent streptavidin. We observed bright psoralen signals on many transcriptionally active interbands and puffs. Upon heat shock, the signals appeared on heat-shock puffs. The signals were resistant to RNase treatment but disappeared or became faint by previous nicking of DNA or inhibition of transcription with {alpha}-amanitin. These data show that transcription-coupled, unconstrained negative supercoils of DNA exist in approximately 150 loci within the interphase genome.

Key words: Drosophila, Polytene Chromosome, Psoralen, Supercoil, Transcription




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[Abstract] [Full Text] [PDF]


© The Company of Biologists Ltd 2004