期刊
STRUCTURE
卷 16, 期 8, 页码 1166-1174出版社
CELL PRESS
DOI: 10.1016/j.str.2008.04.012
关键词
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资金
- U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-06CH1 1357]
- National Institutes of Health [CA100742 to G.L.V]
- Ruth Kirschstein National Research Service Award fellowship [F32 GM077935-01]
- Microbial Sciences Institute
The constant attack on DNA by endogenous and exogenous agents gives rise to nucleolbase modifications that cause mutations, which can lead to cancer. Visualizing the effects of these lesions on the structure of duplex DNA is key to understanding their biologic consequences. The most definitive method of obtaining such structures, X-ray crystallography, i's troublesome to employ owing to the difficulty of obtaining diffraction-quality crystals of DNA. Here, we present a crystallization system that uses a protein, the DNA glycosylase AlkA, as a scaffold to mediate the crystallization of lesion-containing duplex DNA. We demonstrate the use of this system to facilitate the rapid structure determination of DNA containing the lesion 8-oxoguanine in several different sequence contexts, and also deoxyinosine and 1,N 6_ ethenoadenine, each stabilized as the corresponding 2'-flouro analog. The structures of 8-oxoguanine provide a correct atomic-level view of this important endogenous lesion in DNA.
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