Research output: Contribution to journal › Article › peer-review
Displacement of Slow-Turnover DNA Glycosylases by Molecular Traffic on DNA. / Yudkina, Anna V.; Endutkin, Anton V.; Diatlova, Eugenia A. et al.
In: Genes, Vol. 11, No. 8, 866, 08.2020, p. 1-21.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Displacement of Slow-Turnover DNA Glycosylases by Molecular Traffic on DNA
AU - Yudkina, Anna V.
AU - Endutkin, Anton V.
AU - Diatlova, Eugenia A.
AU - Moor, Nina A.
AU - Vokhtantsev, Ivan P.
AU - Grin, Inga R.
AU - Zharkov, Dmitry O.
N1 - Publisher Copyright: © 2020 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/8
Y1 - 2020/8
N2 - 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.
AB - 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.
KW - DNA damage
KW - DNA polymerases
KW - DNA repair
KW - facilitated diffusion
KW - molecular traffic
KW - tight protein–DNA complexes
KW - PROTEIN
KW - ENDONUCLEASE-III
KW - XRCC1
KW - tight protein-DNA complexes
KW - POLYMERASE-BETA
KW - AP LYASE ACTIVITY
KW - HUMAN 8-OXOGUANINE-DNA GLYCOSYLASE
KW - SUBSTRATE-SPECIFICITY
KW - REPLICATION
KW - BASE EXCISION-REPAIR
KW - BINDING
UR - http://www.scopus.com/inward/record.url?scp=85089131803&partnerID=8YFLogxK
U2 - 10.3390/genes11080866
DO - 10.3390/genes11080866
M3 - Article
C2 - 32751599
AN - SCOPUS:85089131803
VL - 11
SP - 1
EP - 21
JO - Genes
JF - Genes
SN - 2073-4425
IS - 8
M1 - 866
ER -
ID: 24950199