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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 authored 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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