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Interestingly, ISWI has been shown to have basal nucleosome binding and repositioning activities independent of its association with other complexes; however, the nucleosome repositioning strategy of ISWI appears to change when it is in these different complexes from creating well-spaced arrays to completely random nucleosome spacing.
Consistent with this conclusion are the results of two recent reports re-evaluating the proposed role of the C-terminal DNA binding domains of ISWI and the related chromatin remodeler Chd1 in the nucleosome repositioning activities of these enzymes.
This suggests a difference in the mechanisms of DNA translocation by ISWI and RSC that might contribute to the differences in the proposed models of their nucleosome repositioning activities.
Indeed, it has been demonstrated that noncatalytic proteins within the ISW2 complex contact the flanking DNA and that those contacts can extend as far as 53 bp and that additional subunits within the ACF complex and the CHRAC complex appear to modulate its nucleosome binding and repositioning activities.
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Initially we characterized the repositioning activity of ISWI using a double-fluorophore-labeled F18N18F substrate.
The ability of chromatin remodelers to translocate along DNA is fundamental to their nucleosome repositioning activity.
Such information is critical for the proper modeling of the nucleosome repositioning activity of ISWI [the following paper (DOI: 10.1021/bi500226b)].
The ability of ISWI to translocate along DNA in an ATP-dependent manner is necessary for its nucleosome repositioning activity.
This ability to translocate along DNA has been shown to be critical to their nucleosome repositioning activity.
ISWI is able to translocate along both single- and double-stranded DNA, a trait necessary for its nucleosome repositioning activity.
Future studies elucidating the mechanism by which these subunits regulate the repositioning activity of ISWI would be of great interest.
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