Human Rev1 relies on insert-2 to promote selective binding and accurate replication of stabilized G-quadruplex motifs
| العنوان: | Human Rev1 relies on insert-2 to promote selective binding and accurate replication of stabilized G-quadruplex motifs |
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| المؤلفون: | Justin W.C. Leung, Jessica H. Hartman, Makayla Berry, Robert L. Eoff, Lane Smith, Alyssa Richey, Julie Gunderson, Leena Maddukuri, Callie Johnson, Megan R. Reed, Amit Ketkar |
| المصدر: | Nucleic Acids Research |
| بيانات النشر: | Oxford University Press, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | DNA Replication, Models, Molecular, AcademicSubjects/SCI00010, Mutant, Genes, myc, DNA-Directed DNA Polymerase, Biology, Genome Integrity, Repair and Replication, medicine.disease_cause, G-quadruplex, Cell Line, 03 medical and health sciences, chemistry.chemical_compound, Genetics, medicine, Humans, Mutation frequency, Nucleotide Motifs, 030304 developmental biology, Alanine, 0303 health sciences, Mutation, 030302 biochemistry & molecular biology, Mutagenesis, DNA, Nucleotidyltransferases, Cell biology, G-Quadruplexes, chemistry, REV1, Protein Binding |
| الوصف: | We previously reported that human Rev1 (hRev1) bound to a parallel-stranded G-quadruplex (G4) from the c-MYC promoter with high affinity. We have extended those results to include other G4 motifs, finding that hRev1 exhibited stronger affinity for parallel-stranded G4 than either anti-parallel or hybrid folds. Amino acids in the αE helix of insert-2 were identified as being important for G4 binding. Mutating E466 and Y470 to alanine selectively perturbed G4 binding affinity. The E466K mutant restored wild-type G4 binding properties. Using a forward mutagenesis assay, we discovered that loss of hRev1 increased G4 mutation frequency >200-fold compared to the control sequence. Base substitutions and deletions occurred around and within the G4 motif. Pyridostatin (PDS) exacerbated this effect, as the mutation frequency increased >700-fold over control and deletions upstream of the G4 site more than doubled. Mutagenic replication of G4 DNA (±PDS) was partially rescued by wild-type and E466K hRev1. The E466A or Y470A mutants failed to suppress the PDS-induced increase in G4 mutation frequency. These findings have implications for the role of insert-2, a motif conserved in vertebrates but not yeast or plants, in Rev1-mediated suppression of mutagenesis during G4 replication. |
| اللغة: | English |
| تدمد: | 1362-4962 0305-1048 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3c08dd83219609f6a81157da30073a87Test http://europepmc.org/articles/PMC7913688Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....3c08dd83219609f6a81157da30073a87 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13624962 03051048 |
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