Diepoxybutane and Diepoxyoctane Interstrand Cross-Linking of the 5S DNA Nucleosomal Core Particle

Diepoxyalkanes form interstrand cross-links in DNA oligomers preferentially at 5‘-GNC sites. We have examined cross-linking by 1,2,3,4-diepoxybutane (DEB) and 1,2,7,8-diepoxyoctane (DEO) within a fragment of the 5S RNA gene of Xenopus borealis in both the free and nucleosomal states. Sites and effic...

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Bibliographic Details
Published in:Biochemistry (Easton) 2001-09, Vol.40 (35), p.10677-10685
Main Authors: Millard, Julie T, Wilkes, Erin E
Format: Article
Language:English
Subjects:
1,2,3,4-diepoxybutane
1,2,7,8-Diepoxyoctane
Animals
Base Sequence
Chickens
Cross-Linking Reagents
DNA - chemistry
Epoxy Compounds - chemistry
erythritol anhydride
Molecular Sequence Data
RNA, Ribosomal, 5S - chemistry
rRNA 5S
Xenopus
Xenopus borealis
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Summary:Diepoxyalkanes form interstrand cross-links in DNA oligomers preferentially at 5‘-GNC sites. We have examined cross-linking by 1,2,3,4-diepoxybutane (DEB) and 1,2,7,8-diepoxyoctane (DEO) within a fragment of the 5S RNA gene of Xenopus borealis in both the free and nucleosomal states. Sites and efficiencies of interstrand cross-linking were probed through denaturing polyacrylamide gel electrophoresis and quantitative phosphorimagery. Both agents targeted 5‘-GNC sites for cross-linking in the restriction fragment in its free state, and DEO also targeted 5‘-GNNC sites. Monoalkylation occurred at all deoxyguanosines. The sites for both monoalkylation and interstrand cross-linking were similar in nucleosomal and free DNA, and cross-linked DNA was cleanly incorporated into the core particle structure. These findings suggest that the 5S core particle is able to tolerate any structural abnormalities induced by diepoxide cross-linking.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi0109663