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Human CCA-adding enzyme is able to repair damaged CCA ends of tRNAs.

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posted on 29.08.2013, 07:57 by Andreas Czech, Sandra Wende, Mario Mörl, Tao Pan, Zoya Ignatova

(A) Total HeLa tRNAs and (B) internally radioactively labeled yeast tRNAPhe were incubated successively with angiogenin and human CCA-adding enzyme. Subsequently to the angiogenin treatment (4 h), T4 polynucleotide kinase (PNK) (45 min) was added which converts the 2′,3′-cyclophosphate ends [42] generated by the angiogenin cleavage to free 3′OH. After purification tRNAs were subjected to treatment with the CCA-adding enzyme (30 min). The 3′-CCA end integrity of the HeLa tRNAs was determined with the fluorescent oligonucleotide (Figure 1A, schematic inset). tRNAPhe lacking the terminal 3′-adenosine (tRNAPheCC) served as a control. The numbers on the left denote the DNA ladder in nt.


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