Detection of OG:A Lesion Mispairs by MutY Relies on a Single His Residue and the 2-Amino Group of 8-Oxoguanine

Lee, Andrea J.; Majumdar, Chandrima; Kathe, Scott D.; Ostrand, Robert P. Van; Vickery, Holly R.; Averill, April M.; Nelson, Shane R.; Manlove, Amelia H.; McCord, Morgan A.; David, Sheila S.

doi:10.1021/jacs.0c04284

Cited by 15 publications

(31 citation statements)

References 21 publications

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“…More recent experiments using higher sampling rates [58] confirmed that the main mode of diffusion for OGG1 on DNA is helical sliding. Single molecule studies on the bacterial Nth, Nei, Fpg, and MutY have recently showed that these DNA glycosylases involved in the repair of oxidative DNA damage also scan for their cognate lesions at rates consistent with random rotational diffusion along the DNA backbone [59,60]. Interestingly, the values of activation energy obtained using instantaneous diffusion rate results are consistent with OGG1 examining flipped-out bases one by one.…”

Section: -Oxog Recognition By Ogg1 On Naked Dnamentioning

confidence: 65%

Lost in the Crowd: How Does Human 8-Oxoguanine DNA Glycosylase 1 (OGG1) Find 8-Oxoguanine in the Genome?

D’Augustin

Huet

Campalans

et al. 2020

IJMS

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The most frequent DNA lesion resulting from an oxidative stress is 7,8-dihydro-8-oxoguanine (8-oxoG). 8-oxoG is a premutagenic base modification due to its capacity to pair with adenine. Thus, the repair of 8-oxoG is critical for the preservation of the genetic information. Nowadays, 8-oxoG is also considered as an oxidative stress-sensor with a putative role in transcription regulation. In mammalian cells, the modified base is excised by the 8-oxoguanine DNA glycosylase (OGG1), initiating the base excision repair (BER) pathway. OGG1 confronts the massive challenge that is finding rare occurrences of 8-oxoG among a million-fold excess of normal guanines. Here, we review the current knowledge on the search and discrimination mechanisms employed by OGG1 to find its substrate in the genome. While there is considerable data from in vitro experiments, much less is known on how OGG1 is recruited to chromatin and scans the genome within the cellular nucleus. Based on what is known of the strategies used by proteins searching for rare genomic targets, we discuss the possible scenarios allowing the efficient detection of 8-oxoG by OGG1.

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Section: -Oxog Recognition By Ogg1 On Naked Dnamentioning

confidence: 65%

Lost in the Crowd: How Does Human 8-Oxoguanine DNA Glycosylase 1 (OGG1) Find 8-Oxoguanine in the Genome?

D’Augustin

Huet

Campalans

et al. 2020

IJMS

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“…We recently showed that MutY utilizes the major groove 2-amino group of OG as a key detection and recognition feature of OG:A mismatches. 26,27 The results presented in this work demonstrate the fidelity of MutY for binding and excising only A, or structures closely resembling A. The fact that T:A or OG:C bps do not present a 2-amino group in the major groove of DNA allows MutY to rapidly bypass these bps and avoid aberrant excision.…”

Section: T H I S C O N T E N T Imentioning

confidence: 66%

“…In previous studies directed at understanding the structural requirements of OG on lesion recognition and catalysis, we showed that MutY relies on the exocyclic 2-amino group of OG to identify and distinguish this mispair from structurally similar T:A bps. 26,27 We also showed that OG binding induces conformational changes that influence A excision. 26,28 Herein, we use structure−activity relationships (SARs) to identify the structural features of A that influence OG:A recognition, verification, base excision, and overall cellular repair.…”

Section: ■ Introductionmentioning

confidence: 95%

Unique Hydrogen Bonding of Adenine with the Oxidatively Damaged Base 8-Oxoguanine Enables Specific Recognition and Repair by DNA Glycosylase MutY

et al. 2020

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The DNA glycosylase MutY prevents deleterious mutations resulting from guanine oxidation by recognition and removal of adenine (A) misincorporated opposite 8-oxo-7,8-dihydroguanine (OG). Correct identification of OG:A is crucial to prevent improper and detrimental MutY-mediatedadenine excision from G:A or T:A base pairs. Here we present a structure–activity relationship (SAR) study using analogues of A to probe the basis for OG:A specificity of MutY. We correlate observed in vitro MutY activity on A analogue substrates with their experimental and calculated acidities to provide mechanistic insight into the factors influencing MutY base excision efficiency. These data show that H-bonding and electrostatic interactions of the base within the MutY active site modulate the lability of the N-glycosidic bond. A analogues that were not excised from duplex DNA as efficiently as predicted by calculations provided insight into other required structural features, such as steric fit and H-bonding within the active site for proper alignment with MutY catalytic residues. We also determined MutY-mediated repair of A analogues paired with OG within the context of a DNA plasmid in bacteria. Remarkably, the magnitudes of decreased in vitro MutY excision rates with different A analogue duplexes do not correlate with the impact on overall MutY-mediated repair. The feature that most strongly correlated with facile cellular repair was the ability of the A analogues to H-bond with the Hoogsteen face of OG. Notably, base pairing of A with OG uniquely positions the 2-amino group of OG in the major groove and provides a means to indirectly select only these inappropriately placed adenines for excision. This highlights the importance of OG lesion detection for efficient MutY-mediated cellular repair. The A analogue SARs also highlight the types of modifications tolerated by MutY and will guide the development of specific probes and inhibitors of MutY.

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“…Adenine excision is known to occur within the N-terminal catalytic domain of MUTYH, while 8-oxoG recognition resides primarily in a C-terminal 8-oxoG recognition domain ( 14–16 ). In a recent study, using single molecule microscopy, in vitro glycosylase and cellular repair assays, the Lee and David laboratories showed that Escherichia coli MutY detects 8-oxoG:A mispairs via the unique major groove positioning of the 2-amino group 8-oxoG using a conserved histidine residue within an ‘FSH’ loop of the C-terminal 8-oxoG recognition domain ( 13 ). Notably, identification of this distinct chemical signal of 8-oxoG:A mismatches would be anticipated to be further complicated in eukaryotic genomes, where the damaged mismatch can be sequestered within chromatin.…”

Section: Introductionmentioning

confidence: 99%

Single molecule analysis indicates stimulation of MUTYH by UV-DDB through enzyme turnover

Jang

Schaich

Khuu

et al. 2021

Nucleic Acids Research

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The oxidative base damage, 8-oxo-7,8-dihydroguanine (8-oxoG) is a highly mutagenic lesion because replicative DNA polymerases insert adenine (A) opposite 8-oxoG. In mammalian cells, the removal of A incorporated across from 8-oxoG is mediated by the glycosylase MUTYH during base excision repair (BER). After A excision, MUTYH binds avidly to the abasic site and is thus product inhibited. We have previously reported that UV-DDB plays a non-canonical role in BER during the removal of 8-oxoG by 8-oxoG glycosylase, OGG1 and presented preliminary data that UV-DDB can also increase MUTYH activity. In this present study we examine the mechanism of how UV-DDB stimulates MUTYH. Bulk kinetic assays show that UV-DDB can stimulate the turnover rate of MUTYH excision of A across from 8-oxoG by 4–5-fold. Electrophoretic mobility shift assays and atomic force microscopy suggest transient complex formation between MUTYH and UV-DDB, which displaces MUTYH from abasic sites. Using single molecule fluorescence analysis of MUTYH bound to abasic sites, we show that UV-DDB interacts directly with MUTYH and increases the mobility and dissociation rate of MUTYH. UV-DDB decreases MUTYH half-life on abasic sites in DNA from 8800 to 590 seconds. Together these data suggest that UV-DDB facilitates productive turnover of MUTYH at abasic sites during 8-oxoG:A repair.

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Detection of OG:A Lesion Mispairs by MutY Relies on a Single His Residue and the 2-Amino Group of 8-Oxoguanine

Cited by 15 publications

References 21 publications

Lost in the Crowd: How Does Human 8-Oxoguanine DNA Glycosylase 1 (OGG1) Find 8-Oxoguanine in the Genome?

Lost in the Crowd: How Does Human 8-Oxoguanine DNA Glycosylase 1 (OGG1) Find 8-Oxoguanine in the Genome?

Unique Hydrogen Bonding of Adenine with the Oxidatively Damaged Base 8-Oxoguanine Enables Specific Recognition and Repair by DNA Glycosylase MutY

Single molecule analysis indicates stimulation of MUTYH by UV-DDB through enzyme turnover

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