The catalytic activity of REV1 is employed during immunoglobulin gene diversification in DT40

Ross, Anna Laura; Sale, Julian E.

doi:10.1016/j.molimm.2005.09.017

Cited by 75 publications

(69 citation statements)

References 39 publications

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“…This is supported by studies suggesting that MMR might act preferentially on the top strand displacing BER to the bottom strand (34,35). Because no significant change was detected in the mutation frequency of C sites (especially in C-to-G mutations) or in G-to-C mutations, which could be contributed by the error-prone polymerase Rev1 (36,37), it seems unlikely that Rev1 activity is impaired by the loss of PCNA ubiquitylation or that Rev1 is responsible for the increase of mutations at C:G pairs in the Pcna Ϫ/Ϫ tg K164R mice. Therefore, other error-prone polymerases may be favored in the absence of ubiquitylation of PCNA and contribute to the strand bias in mutations at C:G residues.…”

Section: Discussionmentioning

confidence: 74%

Ubiquitylated PCNA plays a role in somatic hypermutation and class-switch recombination and is required for meiotic progression

Roa¹,

Avdievich²,

Peled³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

101

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Somatic hypermutation (SHM) and class-switch recombination (CSR) of Ig genes are dependent upon activation-induced cytidine deaminase (AID)-induced mutations. The scaffolding properties of proliferating cell nuclear antigen (PCNA) and ubiquitylation of its residue K164 have been suggested to play an important role organizing the error-prone repair events that contribute to the AID-induced diversification of the Ig locus. We generated knockout mice for PCNA (Pcna ؊/؊ ), which were embryonic lethal. Expression of PCNA with the K164R mutation rescued the lethal phenotype, but the mice (Pcna ؊/؊ tg K164R ) displayed a meiotic defect in early pachynema and were sterile. B cells proliferated normally in Pcna ؊/؊ tg K164R mice, but a PCNA-K164R mutation resulted in impaired ex vivo CSR to IgG1 and IgG3, which was associated with reduced mutation frequency at the switch regions and a bias toward blunt junctions. Analysis of the heavy chain V186.2 region after NP-immunization showed in Pcna ؊/؊ tg K164R mice a significant reduction in the mutation frequency of A:T residues in WA motifs preferred by polymerase-(Pol ), and a strand-biased increase in the mutation frequency of G residues, preferentially in the context of AID-targeted GYW motifs. The phenotype of Pcna ؊/؊ tg K164R mice supports the idea that ubiquitylation of PCNA participates directly in the meiotic process and the diversification of the Ig locus through class-switch recombination (CSR) and somatic hypermutation (SHM).T o mount an effective antibody response, mice and humans create a highly diverse repertoire of antigen binding sites through the rearrangement of the germ line variable (V), diversity (D), and joining (J) Ig locus. Following interaction with antigen, B cells in the germinal centers (GCs) of secondary lymphoid organs express activation-induced cytidine deaminase (AID). AID, together with other enzymes, causes a very high rate (10 Ϫ5 -10 Ϫ3 /base pair/generation) of point mutations in Ig V regions resulting in the affinity maturation and the changes in fine specificity required to produce protective antibodies (1, 2). AID also initiates class-switch recombination (CSR) by mutating the switch regions (SRs) that are located just 5Ј of the constant region genes (3, 4). CSR allows antibodies to be distributed throughout the body and to carry out a wide variety of effector functions. AID deaminates deoxycytidines (dC) in single-stranded DNA in the V and SRs to generate deoxyuridine (dU) (1, 2). However, more than half of the mutations in the V and SRs of mice and humans are in A:T bases and are not the result of the direct biochemical action of AID. Rather, these mutations arise during a second phase of SHM and result from the error-prone base excision repair (BER) and mismatch repair (MMR), both of which are recruited to the dU:dG mismatch generated by AID (1, 2, 4).When critical MMR genes are deleted from mice, most of the mutations in A:T in the V region no longer occur, suggesting that MMR is responsible for the majority of the mutations that ar...

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Section: Discussionmentioning

confidence: 74%

Ubiquitylated PCNA plays a role in somatic hypermutation and class-switch recombination and is required for meiotic progression

Roa¹,

Avdievich²,

Peled³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

101

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“…Independent observations, however, show that human REV1 lacking its BRCT domain can be localized to the nucleus and form foci in unirradiated cells [49]. The BRCT domain of chicken REV1 is not required for DNA damage tolerance [51].…”

Section: Dna Pol ζ In Saccharomyces Cerevisiaementioning

confidence: 99%

“…In mice and in chicken DT40 B-cells, the dCMP transferase activity of Rev1 appears to be important for immunoglobulin diversification. Mutational analysis of immunoglobulin chains indicated that loss of the catalytic activity of Rev1 shifted nucleotide incorporation from C to A or T [46,51].…”

Section: Consequences Of Rev3l Disruption In Higher Organismsmentioning

confidence: 99%

DNA polymerase zeta (pol ζ) in higher eukaryotes

et al. 2007

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Most current knowledge about DNA polymerase zeta (pol ζ) comes from studies of the enzyme in the budding yeast Saccharomyces cerevisiae, where pol ζ consists of a complex of the catalytic subunit Rev3 with Rev7, which associates with Rev1. Most spontaneous and induced mutagenesis in yeast is dependent on these gene products, and yeast pol ζ can mediate translesion DNA synthesis past some adducts in DNA templates. Study of the homologous gene products in higher eukaryotes is in a relatively early stage, but additional functions for the eukaryotic proteins are already apparent. Suppression of vertebrate REV3L function not only reduces induced point mutagenesis but also causes larger-scale genome instability by raising the frequency of spontaneous chromosome translocations. Disruption of Rev3L function is tolerated in Drosophila, Arabidopsis, and in vertebrate cell lines under some conditions, but is incompatible with mouse embryonic development. Functions for REV3L and REV7(MAD2B) in higher eukaryotes have been suggested not only in translesion DNA synthesis but also in some forms of homologous recombination, repair of interstrand DNA crosslinks, somatic hypermutation of immunoglobulin genes and cell-cycle control. This review discusses recent developments in these areas.

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“…Among the four Y-family polymerases, REV1 is unique in its ability to act as a 2Ј-deoxycytidyl transferase (22). Although this transferase activity is suspected to play a role in TLS of abasic residues during somatic hypermutation of immunoglobulin genes (23,24), it does not appear to be important for TLS of other types of DNA damage. This is because (i) the insertion of dC rarely occurs opposite UV-induced lesions, although REV1 is vital in UV mutagenesis, (ii) a catalytically inactive mutant can still support TLS (25), and (iii) a catalytically active mutant cannot always support UV mutagenesis (26).…”

mentioning

confidence: 99%

Two Distinct Translesion Synthesis Pathways across a Lipid Peroxidation-derived DNA Adduct in Mammalian Cells

Yang

Hashimoto

Wind

et al. 2009

Journal of Biological Chemistry

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Translesion DNA synthesis (TLS) of damaged DNA templates is catalyzed by specialized DNA polymerases. To probe the cellular TLS mechanism, a host-vector system consisting of mouse fibroblasts and a replicating plasmid bearing a single DNA adduct was developed. This system was used to explore the TLS mechanism of a heptanone-etheno-dC (H-⑀dC) adduct, an endogenous lesion produced by lipid peroxidation. In wild-type cells, H-⑀dC almost exclusively directed incorporation of dT and dA. Whereas knockout of the Y family TLS polymerase genes, Polh, Polk, or Poli, did not qualitatively affect these TLS events, inactivation of the Rev3 gene coding for a subunit of polymerase or of the Rev1 gene abolished TLS associated with dA, but not dT, insertion. The analysis of results of the cellular studies and in vitro TLS studies using purified polymerases has revealed that the insertion of dA and dT was catalyzed by different polymerases in cells. While insertion of dT can be catalyzed by polymerase , , and , insertion of dA is catalyzed by an unidentified polymerase that cannot catalyze extension from the resulting dA terminus. Therefore, the extension from this terminus requires the activity of polymerase -REV1. These results provide new insight into how cells use different TLS pathways to overcome a synthesis block.Endogenous reactive chemicals such as reactive oxygen species and certain lipid peroxidation products are thought to contribute significantly to aging, age-related degenerative diseases, and cancer (1-4). Cellular DNA is one of their targets, and replication of un-repaired DNA damage introduces mutations into the genome, thereby contributing to the aforementioned biological effects. Heptanone-etheno-dC (H-⑀dC) 2 (Fig. 1) is one of the substituted etheno-base DNA adducts generated by 4-oxo-2(E)-nonenal, a bifunctional electrophilic lipid peroxidation product derived from both arachidonic acid and linoleic acid (5-8). This adduct, as well as H-⑀dG, serves as a biomarker of lipid peroxidation-mediated DNA damage (5). H-⑀dC and H-⑀dG were both found in the DNA of polyps from a cyclooxygenase-2 up-regulated Min mouse, a colorectal cancer mouse model (9). Our previous study has shown that H-⑀dC blocks DNA synthesis and highly miscodes in human cells (10), raising the possibility that cyclooxygenase-2-mediated lipid peroxidation contributes to colorectal carcinogenesis in Min mice through the formation of DNA adducts. Due to its strong genotoxicity and physiological significance, H-⑀dC is an attractive DNA lesion for the mechanistic study of mammalian translesion DNA synthesis (TLS).Recently, it was revealed that various specialized DNA polymerases are actively engaged in DNA synthesis across a DNA lesion (11,12). The catalytic sites of these polymerases are much more spacious than those of replicative polymerases so that they can accommodate a modified template base and an incoming nucleotide (13-15). Accordingly, their fidelity of DNA synthesis is compromised on undamaged template DNA (16 -19). These specialized polyme...

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The catalytic activity of REV1 is employed during immunoglobulin gene diversification in DT40

Cited by 75 publications

References 39 publications

Ubiquitylated PCNA plays a role in somatic hypermutation and class-switch recombination and is required for meiotic progression

Ubiquitylated PCNA plays a role in somatic hypermutation and class-switch recombination and is required for meiotic progression

DNA polymerase zeta (pol ζ) in higher eukaryotes

Two Distinct Translesion Synthesis Pathways across a Lipid Peroxidation-derived DNA Adduct in Mammalian Cells

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