Displacement of Slow-Turnover DNA Glycosylases by Molecular Traffic on DNA

Anna V. Yudkina, Anton V. Endutkin, Eugenia A. Diatlova, Nina A. Moor, Ivan P. Vokhtantsev, Inga R. Grin, Dmitry O. Zharkov

Research output: Contribution to journalArticlepeer-review

Abstract

In the base excision repair pathway, the initiating enzymes, DNA glycosylases, remove damaged bases and form long-living complexes with the abasic DNA product, but can be displaced by AP endonucleases. However, many nuclear proteins can move along DNA, either actively (such as DNA or RNA polymerases) or by passive one-dimensional diffusion. In most cases, it is not clear whether this movement is disturbed by other bound proteins or how collisions with moving proteins affect the bound proteins, including DNA glycosylases. We have used a two-substrate system to study the displacement of human OGG1 and NEIL1 DNA glycosylases by DNA polymerases in both elongation and diffusion mode and by D4, a passively diffusing subunit of a viral DNA polymerase. The OGG1-DNA product complex was disrupted by DNA polymerase β (POLβ) in both elongation and diffusion mode, Klenow fragment (KF) in the elongation mode and by D4. NEIL1, which has a shorter half-life on DNA, was displaced more efficiently. Hence, both possibly specific interactions with POLβ and nonspecific collisions (KF, D4) can displace DNA glycosylases from DNA. The protein movement along DNA was blocked by very tightly bound Cas9 RNA-targeted nuclease, providing an upper limit on the efficiency of obstacle clearance.

Original languageEnglish
Article number866
Pages (from-to)1-21
Number of pages21
JournalGenes
Volume11
Issue number8
DOIs
Publication statusPublished - Aug 2020

Keywords

  • DNA damage
  • DNA polymerases
  • DNA repair
  • facilitated diffusion
  • molecular traffic
  • tight protein–DNA complexes
  • PROTEIN
  • ENDONUCLEASE-III
  • XRCC1
  • tight protein-DNA complexes
  • POLYMERASE-BETA
  • AP LYASE ACTIVITY
  • HUMAN 8-OXOGUANINE-DNA GLYCOSYLASE
  • SUBSTRATE-SPECIFICITY
  • REPLICATION
  • BASE EXCISION-REPAIR
  • BINDING

Fingerprint

Dive into the research topics of 'Displacement of Slow-Turnover DNA Glycosylases by Molecular Traffic on DNA'. Together they form a unique fingerprint.

Cite this