A DNAzyme requiring two different metal ions at two distinct sites

dc.contributor.authorZhou, Wenhu
dc.contributor.authorZhang, Yupei
dc.contributor.authorHuang, Po-Jung Jimmy
dc.contributor.authorDing, Jinsong
dc.contributor.authorLiu, Juewen
dc.date.accessioned2017-04-28T16:12:01Z
dc.date.available2017-04-28T16:12:01Z
dc.date.issued2015-12-10
dc.description.abstractMost previously reported RNA-cleaving DNAzymes require only a single divalent metal ion for catalysis. We recently reported a general trivalent lanthanide-dependent DNAzyme named Ce13d. This work shows that Ce13d requires both Na+ and a trivalent lanthanide (e.g. Ce3+), simultaneously. This discovery is facilitated by the sequence similarity between Ce13d and a recently reported Na+-specific DNAzyme, NaA43. The Ce13d cleavage rate linearly depends on the concentration of both metal ions. Sensitized Tb3+ luminescence and DMS footprinting experiments indicate that the guanines in the enzyme loop are important for Na+-binding. The Na+ dissociation constants of Ce13d measured from the cleavage activity assay, Tb3+ luminescence and DMS footprinting are 24.6, 16.3 and 47 mM, respectively. Mutation studies indicate that the role of Ce3+ might be replaced by G(23) in NaA43. Ce3+ functions by stabilizing the transition state phosphorane, thus promoting cleavage. G(23) competes favorably with low concentration Ce3+ (below 1 mu M). The G(23)-to-hypoxanthine mutation suggests the N1 position of the guanine as a hydrogen bond donor. Together, Ce13d has two distinct metal binding sites, each fulfilling a different role. DNAzymes can be quite sophisticated in utilizing metal ions for catalysis and molecular recognition, similar to protein metalloenzymes.en
dc.description.sponsorshipNatural Sciences and Engineering Research Council of Canada (NSERC); Foundation for Shenghua Scholar of Central South University; National Natural Science Foundation of China [ 21301195]; Fellowship from the China Scholarship Council (CSC) [ 201406370116 to W.Z.]. Funding for open access charge: NSERC.en
dc.identifier.urihttp://dx.doi.org/10.1093/nar/gkv1346
dc.identifier.urihttp://hdl.handle.net/10012/11800
dc.language.isoenen
dc.publisherOxford University Pressen
dc.rightsAttribution-NonCommercial 4.0 International*
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/*
dc.subjectLanthanide-Dependent Dnazymeen
dc.subjectIn-Vitro Selectionen
dc.subjectCleaving DNA Enzymeen
dc.subjectHammerhead Ribozymeen
dc.subjectCleavage Reactionen
dc.subjectRNAen
dc.subjectAciden
dc.subjectEvolutionen
dc.subjectCatalysisen
dc.subjectAptamersen
dc.titleA DNAzyme requiring two different metal ions at two distinct sitesen
dc.typeArticleen
dcterms.bibliographicCitationZhou, W., Zhang, Y., Huang, P.-J. J., Ding, J., & Liu, J. (2016). A DNAzyme requiring two different metal ions at two distinct sites. Nucleic Acids Research, 44(1), 354–363. https://doi.org/10.1093/nar/gkv1346en
uws.contributor.affiliation1Faculty of Scienceen
uws.contributor.affiliation2Chemistryen
uws.contributor.affiliation3Waterloo Institute for Nanotechnology (WIN)en
uws.peerReviewStatusRevieweden
uws.scholarLevelFacultyen
uws.typeOfResourceTexten

Files

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
gkv1346.pdf
Size:
4.77 MB
Format:
Adobe Portable Document Format

License bundle

Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
4.46 KB
Format:
Plain Text
Description: