Human apurinic/apyrimidinic endonuclease 1 is modified in vitro by poly(ADP-ribose) polymerase 1 under control of the structure of damaged DNA
| Autor: | Olga I. Lavrik, Nikita A. Kuznetsov, N. A. Moor, Inna A. Vasil'eva |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
DNA Repair DNA polymerase DNA repair Poly ADP ribose polymerase Poly (ADP-Ribose) Polymerase-1 Biochemistry 03 medical and health sciences chemistry.chemical_compound XRCC1 DNA-(Apurinic or Apyrimidinic Site) Lyase Escherichia coli AP site Cloning Molecular DNA Polymerase beta Polymerase Adenosine Diphosphate Ribose 030102 biochemistry & molecular biology biology DNA General Medicine Base excision repair X-ray Repair Cross Complementing Protein 1 030104 developmental biology chemistry biology.protein DNA Damage Protein Binding |
| Zdroj: | Biochimie. 168:144-155 |
| ISSN: | 0300-9084 |
| DOI: | 10.1016/j.biochi.2019.10.011 |
| Popis: | Apurinic/apyrimidinic endonuclease 1 (APE1) is an essential multifunctional protein in mammals involved in base excision DNA repair (BER), regulation of gene expression and RNA metabolism. Its major enzymatic function is incision of AP sites. Poly(ADP-ribose) polymerase 1 (PARP1) modifies itself and target proteins with poly(ADP-ribose) (PAR), contributing to regulation of many processes. To understand molecular basis of functional cooperation between APE1 and PARP1 in BER, we examined PAR-binding activity and ADP-ribosylation of human APE1 in comparison with known targets of PARP1, using the full-length, N-terminally truncated and catalytically inactive forms of APE1. The protein binds preferentially large ADP-ribose polymers, being very similar to DNA polymerase β (Polβ) but contrasting with the scaffold XRCC1 protein. The interaction with PAR involves the universally conserved catalytic portion and the eukaryote-specific extension of APE1. The ADP-ribosylation of APE1 depends on the structure of PARP1-activating DNA, contrasting APE1 with Polβ and XRCC1. Relative levels of APE1 modification in the presence of different DNA substrates were found to correlate with affinities of the DNAs for APE1 and substrate activities in the enzymatic incision, suggesting the ADP-ribosylation to occur within the DNA-mediated ternary complex. This conclusion was confirmed by importance of the length of DNA region 3' to the AP site for the modification. Deletion of the N-terminal extension of APE1 produced no significant influence on both the ADP-ribosylation efficiency and hydrolytic stability of the modified protein, suggesting localization of target amino acids in the conserved catalytic portion. The most efficient ADP-ribosylation of the catalytically inactive APE1 mutant was shown to reduce the level of PARP1 automodification, suggesting possible role of APE1 in modulating PARP1 activity. Our data on primary role of DNA in controlling the PARP-catalysed modification provide new insights into mechanisms of protein targeting for ADP-ribosylation. |
| Databáze: | OpenAIRE |
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