uracil dna glycosylase Thermo Fisher Search Results


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  • 90
    Thermo Fisher uracil dna glycosylase
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher heat labile
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher uracyl dna glycosylase
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher sybr green supermix uracil dna glycosylase
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher uracyl dna glycosylase udg
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher sybr green qpcr supermix uracil dna glycosylase
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher platinum quantitative pcr supermix uracil n glycosylase udg
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher platinum sybr green qpcr supermix udg
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher platinum sybr green qpcr super mix uracil dna glycosylase kit
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher amperase uracil n glycosylase
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher amperase
    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa <t>ɛC-DNA</t> <t>glycosylase.</t> On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.
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    Thermo Fisher taqman universal master mix ii
    <t>TaqMan</t> RT-qPCR validation results for <t>miR34a</t> show a highly significant (p
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    Thermo Fisher taqman universal pcr master mix no amperase uracil n glycosylase
    <t>TaqMan</t> RT-qPCR validation results for <t>miR34a</t> show a highly significant (p
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    Thermo Fisher gene exp ung hs00422172 m1
    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    Thermo Fisher gene exp smug1 hs00204820 m1
    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    Thermo Fisher gene exp ung mm00449156 m1
    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    Thermo Fisher qpcr reaction
    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    Thermo Fisher gene exp ung mm00449157 g1
    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    Thermo Fisher taqman universal pcr master mix
    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    UBE3B and <t>UNG</t> expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression
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    Thermo Fisher gene exp smug1 mm00452897 m1
    Impact of uracil glycosylase silencing on the mutation profile of Pms2 −/− Ung −/− GC B cells. (A) Analysis of mutations in J H 4 intronic sequences from Pms2 −/− Ung −/− splenic GC B cells (B220 + GL7 + CD95 + ) silenced for Mbd4 , Tdg , <t>Smug1</t> , or both Tdg and Smug1 , as well as Tdg -silenced WT cells. NT represents control cells isolated from bone marrow of Pms2 −/− Ung −/− mice that were cultivated in the same conditions but not transduced before injection in Rag2 −/− recipient mice. KD, knockdown. (B) Pattern of nucleotide substitution for all the genotypes, corrected for base composition. No significant differences for frequencies of GYW/WR(C) or WA/TW hotspot mutations were observed for comparisons of values from individual mice with NT controls (two-tailed Mann Whitney U test). (C) Mutagenesis at A-T base pairs (percentage). Efficiency of knockdown in the analyzed GFP + GC B cells compared with GFP − GC B cells is represented by a color code. Silencing of Tdg was achieved with two different shRNA sequences, represented by dots or triangles. **, P = 0.0012 (TDG KD vs. NT) or 0.0061 (TGD/SMUG KD vs. TDG KD ); two-tailed Mann-Whitney U test. All restored mice (three to seven for each antisense transduction), analyzed individually for glycosylase inhibition and J H 4 mutations, are represented.
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    Image Search Results


    Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa ɛC-DNA glycosylase. On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: A 55-kDa protein isolated from human cells shows DNA glycosylase activity toward 3,N4-ethenocytosine and the G/T mismatch

    doi:

    Figure Lengend Snippet: Silver-stained SDS/PAGE of pooled fractions from purification steps of the HeLa ɛC-DNA glycosylase. On the left are molecular mass markers (BioRad) in kilodaltons, and on the right the arrow indicates the target protein band, which is ≈55-kDa. The gel slice containing this band, after protein elution, denaturation, and renaturation (see Materials and Methods ), showed both ɛC and G/T mismatch cleavage activity.

    Article Snippet: Uracil-DNA glycosylase was obtained from GIBCO/BRL.

    Techniques: Staining, SDS Page, Purification, Activity Assay

    TaqMan RT-qPCR validation results for miR34a show a highly significant (p

    Journal: Scientific Reports

    Article Title: MicroRNA in diagnosis and therapy monitoring of early-stage triple-negative breast cancer

    doi: 10.1038/s41598-018-29917-2

    Figure Lengend Snippet: TaqMan RT-qPCR validation results for miR34a show a highly significant (p

    Article Snippet: The miRNA qPCR reaction was performed for the following housekeeping miRNAs RNU6B and RNU48 and the target miRNA hsa-miR34a-5p with TaqMan Universal Master mix II (without Uracil N-glycosylase) (Applied Biosystems) for 40 cycles using a StepOnePlus (Applied Biosystems) according to manufactures protocol.

    Techniques: Quantitative RT-PCR

    UBE3B and UNG expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression

    Journal: Molecular cancer research : MCR

    Article Title: Alkylation sensitivity screens reveal a conserved cross-species functionome

    doi: 10.1158/1541-7786.MCR-12-0168

    Figure Lengend Snippet: UBE3B and UNG expression fluctuate in cancer cell lines. UNG is expressed at higher levels in brain tumor cell lines, when compared to non-tumor controls. Quantification of UNG and UBE3B mRNA by qRT-PCR compared to human astrocyte controls. Gene expression

    Article Snippet: Applied Biosystems TaqMan® Gene Expression Assays used are as follows: human OGG1: Hs00213454_m1; human SMUG1: Hs00204820_m1; human MBD4: Hs00187498_m1; human UNG: Hs00422172_m1; human MYH: Hs01014856_m1; human NTHL1: Hs00267385; human MPG: Hs01012594_m1; human NEIL1: Hs0022637_m1; human NEIL2: Hs00376746_m1; human NEIL3: Hs00217387_m1; human UBE3B: Hs00296200_m1; Human ICMT: Hs00202655_m1; Human B4GALT7: Hs01011258_m1; Human CHRM3: Hs00265216_s1; Human PADI1: Hs00203458_m1.

    Techniques: Expressing, Quantitative RT-PCR

    Depletion of UNG protein and uracil glycosylase activity in glioma cells. (A) Knockdown of UNG mRNA corresponds with decreased UNG protein levels. Control and UNG-KD cell line nuclear extracts were resolved in a 4-20% SDS/PAGE gel and immunoblotted for

    Journal: Molecular cancer research : MCR

    Article Title: Alkylation sensitivity screens reveal a conserved cross-species functionome

    doi: 10.1158/1541-7786.MCR-12-0168

    Figure Lengend Snippet: Depletion of UNG protein and uracil glycosylase activity in glioma cells. (A) Knockdown of UNG mRNA corresponds with decreased UNG protein levels. Control and UNG-KD cell line nuclear extracts were resolved in a 4-20% SDS/PAGE gel and immunoblotted for

    Article Snippet: Applied Biosystems TaqMan® Gene Expression Assays used are as follows: human OGG1: Hs00213454_m1; human SMUG1: Hs00204820_m1; human MBD4: Hs00187498_m1; human UNG: Hs00422172_m1; human MYH: Hs01014856_m1; human NTHL1: Hs00267385; human MPG: Hs01012594_m1; human NEIL1: Hs0022637_m1; human NEIL2: Hs00376746_m1; human NEIL3: Hs00217387_m1; human UBE3B: Hs00296200_m1; Human ICMT: Hs00202655_m1; Human B4GALT7: Hs01011258_m1; Human CHRM3: Hs00265216_s1; Human PADI1: Hs00203458_m1.

    Techniques: Activity Assay, SDS Page

    Impact of uracil glycosylase silencing on the mutation profile of Pms2 −/− Ung −/− GC B cells. (A) Analysis of mutations in J H 4 intronic sequences from Pms2 −/− Ung −/− splenic GC B cells (B220 + GL7 + CD95 + ) silenced for Mbd4 , Tdg , Smug1 , or both Tdg and Smug1 , as well as Tdg -silenced WT cells. NT represents control cells isolated from bone marrow of Pms2 −/− Ung −/− mice that were cultivated in the same conditions but not transduced before injection in Rag2 −/− recipient mice. KD, knockdown. (B) Pattern of nucleotide substitution for all the genotypes, corrected for base composition. No significant differences for frequencies of GYW/WR(C) or WA/TW hotspot mutations were observed for comparisons of values from individual mice with NT controls (two-tailed Mann Whitney U test). (C) Mutagenesis at A-T base pairs (percentage). Efficiency of knockdown in the analyzed GFP + GC B cells compared with GFP − GC B cells is represented by a color code. Silencing of Tdg was achieved with two different shRNA sequences, represented by dots or triangles. **, P = 0.0012 (TDG KD vs. NT) or 0.0061 (TGD/SMUG KD vs. TDG KD ); two-tailed Mann-Whitney U test. All restored mice (three to seven for each antisense transduction), analyzed individually for glycosylase inhibition and J H 4 mutations, are represented.

    Journal: The Journal of Experimental Medicine

    Article Title: Pms2 and uracil-DNA glycosylases act jointly in the mismatch repair pathway to generate Ig gene mutations at A-T base pairs

    doi: 10.1084/jem.20161576

    Figure Lengend Snippet: Impact of uracil glycosylase silencing on the mutation profile of Pms2 −/− Ung −/− GC B cells. (A) Analysis of mutations in J H 4 intronic sequences from Pms2 −/− Ung −/− splenic GC B cells (B220 + GL7 + CD95 + ) silenced for Mbd4 , Tdg , Smug1 , or both Tdg and Smug1 , as well as Tdg -silenced WT cells. NT represents control cells isolated from bone marrow of Pms2 −/− Ung −/− mice that were cultivated in the same conditions but not transduced before injection in Rag2 −/− recipient mice. KD, knockdown. (B) Pattern of nucleotide substitution for all the genotypes, corrected for base composition. No significant differences for frequencies of GYW/WR(C) or WA/TW hotspot mutations were observed for comparisons of values from individual mice with NT controls (two-tailed Mann Whitney U test). (C) Mutagenesis at A-T base pairs (percentage). Efficiency of knockdown in the analyzed GFP + GC B cells compared with GFP − GC B cells is represented by a color code. Silencing of Tdg was achieved with two different shRNA sequences, represented by dots or triangles. **, P = 0.0012 (TDG KD vs. NT) or 0.0061 (TGD/SMUG KD vs. TDG KD ); two-tailed Mann-Whitney U test. All restored mice (three to seven for each antisense transduction), analyzed individually for glycosylase inhibition and J H 4 mutations, are represented.

    Article Snippet: Relative transcript abundance was assessed by real-time PCR on a PCR machine (7500 Fast System Real-Time; Applied Biosystems; 2 min at 50°C, 10 min at 95°C, and 40 cycles of 10 s at 95°C and 1 min at 60°C) in universal PCR master mix (No AmpErase UNG; Applied Biosystems), with the following validated TaqMan assays: Mm00437762_m1 (β2-microglobulin), Mm01225357_g1 (Tdg), Mm01184338_m1 (Mbd4), and Mm00452897_m1 (Smug1).

    Techniques: Mutagenesis, Isolation, Mouse Assay, Injection, Two Tailed Test, MANN-WHITNEY, shRNA, Transduction, Inhibition

    Analysis of uracil glycosylase expression levels by qRT-PCR. (A) Relative expression levels of Mbd4 , Tdg , and Smug1 in naive B cells (B220 + GL7 − CD95 − ) from spleen of immunized WT ( n = 4) and Ung −/− Pms2 −/− ( n = 3) mice, compared with β-2 microglobulin (b2m). (B) Relative expression of Mbd4 , Tdg , and Smug1 in GC B cells (B220 + GL7 + CD95 + ) compared with naive B cells from spleens of WT ( n = 4) and Ung −/− Pms2 −/− ( n = 3) mice. The dotted line represents a ratio of 1 between GC and naive B cells. Quantification was performed in triplicates for RNA samples from three to four different mice. Error bars represent SD.

    Journal: The Journal of Experimental Medicine

    Article Title: Pms2 and uracil-DNA glycosylases act jointly in the mismatch repair pathway to generate Ig gene mutations at A-T base pairs

    doi: 10.1084/jem.20161576

    Figure Lengend Snippet: Analysis of uracil glycosylase expression levels by qRT-PCR. (A) Relative expression levels of Mbd4 , Tdg , and Smug1 in naive B cells (B220 + GL7 − CD95 − ) from spleen of immunized WT ( n = 4) and Ung −/− Pms2 −/− ( n = 3) mice, compared with β-2 microglobulin (b2m). (B) Relative expression of Mbd4 , Tdg , and Smug1 in GC B cells (B220 + GL7 + CD95 + ) compared with naive B cells from spleens of WT ( n = 4) and Ung −/− Pms2 −/− ( n = 3) mice. The dotted line represents a ratio of 1 between GC and naive B cells. Quantification was performed in triplicates for RNA samples from three to four different mice. Error bars represent SD.

    Article Snippet: Relative transcript abundance was assessed by real-time PCR on a PCR machine (7500 Fast System Real-Time; Applied Biosystems; 2 min at 50°C, 10 min at 95°C, and 40 cycles of 10 s at 95°C and 1 min at 60°C) in universal PCR master mix (No AmpErase UNG; Applied Biosystems), with the following validated TaqMan assays: Mm00437762_m1 (β2-microglobulin), Mm01225357_g1 (Tdg), Mm01184338_m1 (Mbd4), and Mm00452897_m1 (Smug1).

    Techniques: Expressing, Quantitative RT-PCR, Mouse Assay

    A model whereby both uracil glycosylases and the Pms2/Mlh1 complex enable A-T base pair mutagenesis with a distinct strand bias. (A) Overlapping enzymatic activities generate entry sites for ExoI in MMR-mediated A-T mutagenesis. (Right) Recognition of AID deamination products by Msh2–Msh6 stimulates the endonuclease activity of the Pms2–Mlh1 complex, which acts at distance on either DNA strands. (Left) In a situation of processive deamination, nicks introduced by uracil glycosylase–mediated base excision (UNG, SMUG1, and TDG) and APEX2 strand incision can participate in MMR-mediated error-prone repair. Glycosylases may also act in the context of two independent deamination events on both DNA strands or, possibly, in the context of a single uracil (see Discussion). (B) Pms2 versus uracil glycosylase contribution estimated from A/T mutation ratios. The 1.8- to twofold A over T ratio observed in hypermutation was estimated to correspond to a 3:1 strand bias in error-prone DNA synthesis, taking into account the intrinsic T over A mutation preference of Pol η (a 4:1 higher mutation frequency opposite T than opposite A; Zivojnovic et al., 2014 ). This strand bias is represented by arrows of unequal length, representing the relative A:T mutagenic patch on each strand, without aiming at indicating whether the patch is shorter or less frequent on the transcribed strand or differently located on the locus. If one considers, from mutations observed in Pms2 −/− mice, that uracil glycosylases operate mainly on the coding, nontranscribed strand and that PMS2 endonuclease operates without strand bias and is responsible for most mutations introduced in the transcribed strand, both pathways appear to contribute equally to the generation of Msh2/6-driven A-T mutations.

    Journal: The Journal of Experimental Medicine

    Article Title: Pms2 and uracil-DNA glycosylases act jointly in the mismatch repair pathway to generate Ig gene mutations at A-T base pairs

    doi: 10.1084/jem.20161576

    Figure Lengend Snippet: A model whereby both uracil glycosylases and the Pms2/Mlh1 complex enable A-T base pair mutagenesis with a distinct strand bias. (A) Overlapping enzymatic activities generate entry sites for ExoI in MMR-mediated A-T mutagenesis. (Right) Recognition of AID deamination products by Msh2–Msh6 stimulates the endonuclease activity of the Pms2–Mlh1 complex, which acts at distance on either DNA strands. (Left) In a situation of processive deamination, nicks introduced by uracil glycosylase–mediated base excision (UNG, SMUG1, and TDG) and APEX2 strand incision can participate in MMR-mediated error-prone repair. Glycosylases may also act in the context of two independent deamination events on both DNA strands or, possibly, in the context of a single uracil (see Discussion). (B) Pms2 versus uracil glycosylase contribution estimated from A/T mutation ratios. The 1.8- to twofold A over T ratio observed in hypermutation was estimated to correspond to a 3:1 strand bias in error-prone DNA synthesis, taking into account the intrinsic T over A mutation preference of Pol η (a 4:1 higher mutation frequency opposite T than opposite A; Zivojnovic et al., 2014 ). This strand bias is represented by arrows of unequal length, representing the relative A:T mutagenic patch on each strand, without aiming at indicating whether the patch is shorter or less frequent on the transcribed strand or differently located on the locus. If one considers, from mutations observed in Pms2 −/− mice, that uracil glycosylases operate mainly on the coding, nontranscribed strand and that PMS2 endonuclease operates without strand bias and is responsible for most mutations introduced in the transcribed strand, both pathways appear to contribute equally to the generation of Msh2/6-driven A-T mutations.

    Article Snippet: Relative transcript abundance was assessed by real-time PCR on a PCR machine (7500 Fast System Real-Time; Applied Biosystems; 2 min at 50°C, 10 min at 95°C, and 40 cycles of 10 s at 95°C and 1 min at 60°C) in universal PCR master mix (No AmpErase UNG; Applied Biosystems), with the following validated TaqMan assays: Mm00437762_m1 (β2-microglobulin), Mm01225357_g1 (Tdg), Mm01184338_m1 (Mbd4), and Mm00452897_m1 (Smug1).

    Techniques: Mutagenesis, Activity Assay, Activated Clotting Time Assay, DNA Synthesis, Mouse Assay