Mutational and Kinetic Analysis of APE1 Endoribonuclease Activity

Mutational and Kinetic Analysis of APE1 Endoribonuclease Activity Full article

Journal

Molekuliarnaia biologiia

Output data

Year: 2021, Volume: 55, Number: 2, Pages: 243-257 Pages count : 15 DOI: 10.31857/S0026898421020099

Tags

active site; APE1; endoribonuclease activity; human apurine/apyrimidine endonuclease; RNA substrates; site-directed mutagenesis

Authors

Kuznetsova A.A. ¹ , Gavrilova A.A. ^1,2 , Novopashina D.S. ¹ , Fedorova O.S. ¹ , Kuznetsov N.A. ¹

Affiliations

1	Institute of Chemical Biology and Fundamental Medicine, Russian Academy of Sciences, Siberian Branch, Novosibirsk, 630090, Russian Federation
2	Novosibirsk National Research State University, Novosibirsk, 630090, Russian Federation

Human apurinic/apyrimidinic endonuclease 1 (APE1) participates in the DNA repair system. It is believed that the main biological function of APE1 is Mg^(2+)-dependent hydrolysis of AP-sites in DNA. On the base of structural data, kinetic studies, and mutation analysis, the key stages of APE1 interaction with damaged DNA were established. It has been shown recently that APE1 can act as an endoribonuclease that catalyzes mRNA hydrolysis at certain pyrimidine-purine sites and thus controls the level of certain transcripts. In addition, the presence of Mg^(2+) ions was shown to be not required for the endoribonuclease activity of APE1, in contrast to the AP-endonuclease activity. This indicates differences in mechanisms of APE1 catalysis on RNA and DNA substrates, but the reasons for these differences remain unclear. Here, the analysis of endoribonuclease hydrolysis of model RNA substrates with wild type APE1 enzyme and its mutant forms Y171F, R177F, R181A, D210N, N212A, T268D, M270A, and D308A, was performed. It was shown that mutation of Asn212, Asp210, and Tyr171 residues leads to the decrease of AP-endonuclease activity while endoribonuclease activity is retained. Also, T268D and M270A APE1 mutants lose specificity to pyrimidine-purine sequences. R177F and R181A did not show a significant decrease in enzyme activity, whereas D308A demonstrated a decrease of endoribonuclease activity.

Kuznetsova A.A. , Gavrilova A.A. , Novopashina D.S. , Fedorova O.S. , Kuznetsov N.A.
Mutational and Kinetic Analysis of APE1 Endoribonuclease Activity
Molekuliarnaia biologiia. 2021. V.55. N2. P.243-257. DOI: 10.31857/S0026898421020099 WOS Scopus РИНЦ OpenAlex

Кузнецова А.А. , Гаврилова А.А. , Новопашина Д.С. , Федорова О.С. , Кузнецов Н.А.
Мутационный и кинетический анализ эндорибонуклеазной активности APE1
Молекулярная биология. 2021. Т.55. №2. С.243-257. РИНЦ

Web of science:	WOS:000646053000005
Scopus:	2-s2.0-85104587048
Elibrary:	48887840
OpenAlex:	W3160525806

DB	Citing
Scopus	2
Web of science	7
Elibrary	4