Nucleic Acids Research, Vol 26, Issue 14 3364-3371, Copyright © 1998 by Oxford University Press
A Boybek, TD Ray, MC Evans and PJ Dyson
Both Streptomyces lividans and Streptomyces avermitilis encode similar
systems of post-replicative DNA modification which act site- specifically
on closely opposed guanines on either strand. The modifications can be
detected since they react in vitro with an oxidative derivative of Tris,
resulting in strand cleavage. Previous analysis of the preferred
modification site of plasmid pIJ101 indicated that extensive amounts of
flanking sequence, including direct and inverted repeat structures, are
required to direct modification in vivo within a central 6 bp palindrome.
We have now examined the preferred modification sites of a chromosomal
element, the 5.7 kb amplified DNA sequence (ADS5.7) found in certain S.
lividans mutants. In contrast to the pIJ101 site, each of the ADS5. 7sites
is intragenic and modified with a 10-fold reduced frequency. However,
similar extents of flanking sequence are required for authentic
double-strand modification; deletion mutants exhibited different
modification profiles, including displaced double-stranded or
single-stranded modi-fication. Comparison of different modification sites
reveals conservation of the central core sequence, but no significant
similarities between flanking sequences. Enhanced modification was detected
in a cloned region of the ADS5.7, suggesting that local DNA topology,
probably influenced by both DNA supercoiling and the nature of flanking
sequences, can influence the modifying activity.
ARTICLES
Novel site-specific DNA modification in Streptomyces: analysis of preferred intragenic modification sites present in a 5.7 kb amplified DNA sequence
Molecular Biology Research Group, School of Biological Sciences, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK.
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