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TitleDestabilization of the PCNA trimer mediated by its interaction with the NEIL1 DNA glycosylase.
Journal, issue, pagesNucleic Acids Res, Vol. 45, Issue 5, Page 2897-2909, Year 2017
Publish dateMar 17, 2017
AuthorsAishwarya Prakash / Kedar Moharana / Susan S Wallace / Sylvie Doublié /
PubMed AbstractThe base excision repair (BER) pathway repairs oxidized lesions in the DNA that result from reactive oxygen species generated in cells. If left unrepaired, these damaged DNA bases can disrupt ...The base excision repair (BER) pathway repairs oxidized lesions in the DNA that result from reactive oxygen species generated in cells. If left unrepaired, these damaged DNA bases can disrupt cellular processes such as replication. NEIL1 is one of the 11 human DNA glycosylases that catalyze the first step of the BER pathway, i.e. recognition and excision of DNA lesions. NEIL1 interacts with essential replication proteins such as the ring-shaped homotrimeric proliferating cellular nuclear antigen (PCNA). We isolated a complex formed between NEIL1 and PCNA (±DNA) using size exclusion chromatography (SEC). This interaction was confirmed using native gel electrophoresis and mass spectrometry. Stokes radii measured by SEC hinted that PCNA in complex with NEIL1 (±DNA) was no longer a trimer. Height measurements and images obtained by atomic force microscopy also demonstrated the dissociation of the PCNA homotrimer in the presence of NEIL1 and DNA, while small-angle X-ray scattering analysis confirmed the NEIL1 mediated PCNA trimer dissociation and formation of a 1:1:1 NEIL1-DNA-PCNA(monomer) complex. Furthermore, ab initio shape reconstruction provides insights into the solution structure of this previously unreported complex. Together, these data point to a potential mechanistic switch between replication and BER.
External linksNucleic Acids Res / PubMed:27994037 / PubMed Central
MethodsSAS (X-ray synchrotron)
Structure data

SASDBB7:
Human NEI like DNA glycosylase 1 (NEIL1) bound to DNA
Method: SAXS/SANS

SASDBC7:
Human NEI like DNA glycosylase 1 (NEIL1) (Endonuclease 8-like 1)
Method: SAXS/SANS

SASDBD7:
Human Proliferating Cell Nuclear Antigen (PCNA) (Proliferating cell nuclear antigen)
Method: SAXS/SANS

Source
  • Homo sapiens (human)

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