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Record W2913985885 · doi:10.1021/acscatal.8b04889

Exploring the Identity of the General Base for a DNA Polymerase Catalyzed Reaction Using QM/MM: The Case Study of Human Translesion Synthesis Polymerase η

2019· article· en· W2913985885 on OpenAlexafffund
Katie A. Wilson, Pedro Alexandrino Fernandes, Maria J. Ramos, Stacey D. Wetmore

Bibliographic record

VenueACS Catalysis · 2019
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicDNA Repair Mechanisms
Canadian institutionsUniversity of Lethbridge
FundersEuropean Regional Development FundFundação para a Ciência e a TecnologiaNatural Sciences and Engineering Research Council of CanadaCanada Foundation for InnovationAlberta Innovates - Technology FuturesUniversity of Lethbridge
KeywordsPolymeraseDNA polymeraseDNA replicationChemistryONIOMDNA clampBiophysicsDNAStereochemistryBiochemistryBiologyCatalysisPolymerase chain reactionReverse transcriptaseGene

Abstract

fetched live from OpenAlex

DNA replication is a fundamental process for life. Despite previous experimental and computational studies of the reaction pathway catalyzed by a variety of DNA polymerases, certain features of the mechanism remain unclear, especially for the more recently identified translesion synthesis (TLS) polymerases. A pressing question in the TLS polymerase literature is: what is the identity of the general base that initiates the reaction? The current work uses computational techniques to shed light on this topic by exploring the mechanism for the chemical step catalyzed by a human TLS polymerase, namely polymerase η. This enzyme was chosen due to its critical role in the replication of damaged DNA and its biomedical importance. A consistent theoretical framework is used to investigate all possibilities for the general base based on previous proposals from experimental and computational studies on polymerase η and other DNA polymerases. Our MD and ONIOM quantum mechanics/molecular mechanics (QM/MM) calculations suggest that the dNTP α-phosphate moiety and active site water interacting with bulk solvent or the catalytic Mg2+ ion are not optimal choices for the general base due to high energy intermediates. Although proton transfer to an active site hydroxide ion or to Glu116 through Ser113 is energetically feasible, the pathway that best aligns with our dynamical (MD/umbrella sampling) structural information and available experimental kinetic and mutational data invokes Glu116 directly abstracting a proton to initiate the nucleotidyl transfer reaction. Together, these results help answer a long-standing question in the DNA replication literature.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.001
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.005
Threshold uncertainty score0.888

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0010.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.050
GPT teacher head0.290
Teacher spread0.239 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations22
Published2019
Admission routes2
Has abstractyes

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