Enzymatic characterization of translesion synthesis by Sulfolobus acidocaldarius DNA polymerase IV

Abstract:

[Objective] To characterize the translesion synthesis of different damages by DNA polymerase IV from thermophilic archaea Sulfolobus acidocaldarius. [Methods] We detected the translesion synthesis ability of Sulfolobus acidocaldarius polymerase IV (SacpolIV) using fluorescence labeled oligonucleotides as substrates through denaturing polyacrylamide gel electrophoresis. [Results] We successfully purified SacpolIV from induced E. coli and confirmed SacpolIV possessed strong translesion synthesis ability to DNA damages on purines and pyrimidines. The TLS ability depends on whether damaged bases can pair normal bases perfectly. Besides, we found SacpolIV can incorporate rNMP into DNA strand. [Conclusion] We confirmed that SacpolIV is a typical TLS DNA polymerase with the abilities of bypassing damaged purines and pyrimidines on DNA, and incorporating rNMP into DNA.