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. 2016 Dec 15;64(6):1109–1116. doi: 10.1016/j.molcel.2016.11.014

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© 2016 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

TaqToxin/DarT ADP-Ribosylates ssDNA Oligonucleotides on Thymidines with Sequence Preference

(A) Autoradiography of denaturing polyacrylamide gel analyzing TaqToxin ADP-ribosylation modification reactions using short oligonucleotide (GJ1) or its complementary sequence (GJ1rc) as substrates in the presence of ³²P-NAD⁺.

(B) UV detection of ethidium bromide-stained denaturing polyacrylamide gels separating reactions as in (A) in the presence of NAD⁺.

(C) Mass spectra of modified (top) and non-modified (bottom) 9-mer GJ4-Ts oligonucleotide. The double- and triple-charged molecular ions are clearly detected. The shift of m/z values of the molecular ions for the modified oligonucleotide corresponds to ADP-ribosylation.

(D) Diagnostic ion magnification of tandem mass spectrometry (MS/MS) spectra of the oligonucleotides as in (C). Relevant fragments are indicated on the side, while the key fragment is highlighted in orange. The difference between modified and non-modified fragments corresponds to the size of the ADP-ribosyl moiety.