Characterization of biochemical properties of an apurinic/apyrimidinic endonuclease from Helicobacter pylori

Aigerim Turgimbayeva, Sailau Abeldenov, Dmitry O. Zharkov, Alexander A. Ishchenko, Yerlan Ramankulov, Murat Saparbaev, Bekbolat Khassenov

Результат исследования: Научные публикации в периодических изданияхстатья

2 Цитирования (Scopus)

Аннотация

Apurinic/apyrimidinic (AP) endonucleases play critical roles in the repair of abasic sites and strand breaks in DNA. Complete genome sequences of Helicobacter pylori reveal that this bacterial specie has a single AP endonuclease. An H. pylori homolog of Xth (HpXth) is a member of exonuclease III family, which is represented by Escherichia coli Xth. Currently, it remains unknown whether this single AP endonuclease has DNA repair activities similar to those of its counterpart in E. coli and other bacteria. We report that HpXth possesses efficient AP site cleavage, 3′-repair phosphodiesterase, and 3′-phosphatase activities but not the nucleotide incision repair function. Optimal reaction conditions for HpXth's AP endonuclease activity are low ionic strength, high Mg2+ concentration, pH in the range 7-8, and temperature 30°C. The kinetic parameters measured under steady-state conditions showed that HpXth removes the AP site, 3′-blocking sugar-phosphate, and 3′-terminal phosphate in DNA strand breaks with good efficiency (kcat/KM = 1240, 44, and 5,4 μM-1·min-1, respectively), similar to that of E. coli Xth. As expected, the presence of HpXth protein in AP endonuclease Ddeficient E. coli xth nfo strain significantly reduced the sensitivity to an alkylating agent and H2O2. Mutation of active site residue D144 in HpXth predicted to be essential for catalysis resulted in a complete loss of enzyme activities. Several important structural features of HpXth were uncovered by homology modeling and phylogenetic analysis. Our data show the DNA substrate specificity of H. pylori AP endonuclease and suggest that HpXth counteracts the genotoxic effects of DNA damage generated by endogenous and hostimposed factors.

Язык оригиналаанглийский
Номер статьиe0202232
Страницы (с-по)e0202232
Число страниц26
ЖурналPLoS ONE
Том13
Номер выпуска8
DOI
СостояниеОпубликовано - 15 авг 2018

Fingerprint Подробные сведения о темах исследования «Characterization of biochemical properties of an apurinic/apyrimidinic endonuclease from Helicobacter pylori». Вместе они формируют уникальный семантический отпечаток (fingerprint).

Цитировать