DNA Polymerase λ Mediates a Back-up Base Excision Repair Activity in Extracts of Mouse Embryonic Fibroblasts

Braithwaite, Elena K.; Prasad, Rajendra; Shock, David D.; Hou, Esther W.; Beard, William A.; Wilson, Samuel H.

doi:10.1074/jbc.m411864200

Cited by 146 publications

(140 citation statements)

References 39 publications

(45 reference statements)

Supporting

Mentioning

131

Contrasting

Order By: Relevance

“…Recently, it was also demonstrated that in addition to pol β, DNA polymerase λ (pol λ) and DNA polymerase ι (pol ι), as well as the mitochondrial polymerase pol γ have 5′dRP lyase activity and can function (in vitro) in the removal of the 5′ dRP group that arises during BER [53,[57][58][59][60][61]. This would suggest that pol ι and/or pol λ may participate in BER, possibly as a back-up to pol β or may function with unique lesion specificity, in-line with the proposition that the initiating lesion directs the repair sub-pathway [51,62].…”

Section: Nih Public Accessmentioning

confidence: 99%

“…Although MEFs deficient in either pol ι or pol λ are not hypersensitive to alkylating agents [44], the ability of either pol ι and/or pol λ to complement pol β in vivo has not been evaluated. Using MEFs isolated from pol λ knockout mice [63], a back-up role for pol λ was observed following oxidative damage, although the requirement for the 5′dRP activity of pol λ has not been determined [53,59]. To date, a targeted deletion or inactivation of the mouse POLI gene has not been reported.…”

Section: Nih Public Accessmentioning

confidence: 99%

See 1 more Smart Citation

DNA polymerase beta null mouse embryonic fibroblasts harbor a homozygous null mutation in DNA polymerase iota

Sobol

2007

DNA Repair

View full text Add to dashboard Cite

Dear EditorPrior to the explosion of gene targeting technology, considerable biochemical and cell-based experiments suggested a role for DNA polymerase β (pol β) in DNA repair. Since its initial discovery in 1971 [1], the enzyme that was to eventually be designated pol β [2,3] was unique in its enzymatic properties [4][5][6] as compared to the other newly characterized mammalian DNA polymerases alpha (pol α), gamma (pol γ) and delta (polδ) [1,7]. Of the four eukaryotic DNA polymerases identified by 1977, pol β soon earned the moniker of 'the' DNA repair polymerase [8][9][10][11]. These early studies defined a role for pol β in repair using isolated nuclei or nuclear extracts, monitoring the incorporation of radioactive nucleosides following ultraviolet (UV) light-induced DNA damage [8][9][10][11]. A more definitive role for pol β in excision repair in vitro was demonstrated in 1983, in which complete removal of pyrimidine dimers in DNA was conducted by purified enzyme preparations of pol β, T4 denV (UV endonuclease) and HeLa AP endonuclease II [12]. Although it was subsequently shown that pol α can also carry-out gap-filling DNA synthesis in a base excision repair (BER) reaction [13], the evidence continued to mount in support of pol β acting as 'the' DNA repair polymerase in the nucleus, while pol γ participated in replication and repair in mitochondria [1]. Studies continued to identify a role for pol β in the repair of damage induced by many different DNA damaging agents, including bleomycin [14,15] [21][22][23][24]. Several groups reported complete BER in vitro with pol β and additional purified proteins [22,23,25]. Although it was demonstrated in heterologous systems (E. coli and Saccharomyces cerevisiae) that pol β can conduct DNA replication and repair in vivo [26,27], it was not until a mouse gene knockout was made that the specificity of the repair conducted by pol β was defined [28].Characterization of the pol β knockout mouse [29,30] [34,54,55]; among other studies too numerous to mention herein. The most definitive and reproducible endpoint that has been used to evaluate pol β *To whom correspondence should be addressed: Robert W. Sobol, Hillman Cancer Center, University of Pittsburgh Cancer Institute, Research Pavilion, Suite 2.6, 5117 Centre Avenue, Pittsburgh, PA 15213-1863, Phone: 412-623-7764, Fax: 412-623-7761, e-mail: rws9@pitt.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript participation in repair in vivo is survival following DNA damage such as exposure to alkylating agen...

show abstract

Section: Nih Public Accessmentioning

confidence: 99%

Section: Nih Public Accessmentioning

confidence: 99%

DNA polymerase beta null mouse embryonic fibroblasts harbor a homozygous null mutation in DNA polymerase iota

Sobol

2007

DNA Repair

View full text Add to dashboard Cite

show abstract

“…This flexibility was first suggested by the observation that Pol λ generates deletion errors at an extremely high rate [11], and is now thought to be a feature of the enzyme that facilitates its role in vivo. The phenotypes of mice deficient in this polymerase indicate that Pol λ plays a role in the V(D)J process of antigen gene diversification [12], and several reports implicate Pol λ in Base Excision Repair [13][14][15] and the repair of double-strand breaks through the Non-Homologous DNA End-Joining pathway [16][17][18]. It is thus interesting to examine the details of the polymerization reaction as catalyzed by Pol λ in order to understand whether specific catalytic features can account for these special properties.…”

Section: Introductionmentioning

confidence: 99%

Role of the catalytic metal during polymerization by DNA polymerase lambda

et al. 2007

View full text Add to dashboard Cite

The incorporation of dNMPs into DNA by polymerases involves a phosphoryl transfer reaction hypothesized to require two divalent metal ions. Here we investigate this hypothesis using as a model human DNA polymerase λ (Pol λ), an enzyme suggested to be activated in vivo by manganese. We report the crystal structures of four complexes of human Pol λ. In a 1.9Å structure of Pol λ containing a 3′ OH and the non-hydrolyzable analog dUpnpp, a non-catalytic Na + ion occupies the site for metal A and the ribose of the primer-terminal nucleotide is found in a conformation that positions the acceptor 3'OH out of line with the α-phosphate and the bridging oxygen of the pyrophosphate leaving group. Soaking this crystal in MnCl 2 yielded a 2.0Å structure with Mn 2+ occupying the site for metal A. In the presence of Mn 2+ , the conformation of the ribose is C3' endo and the 3'-oxygen is in line with the leaving oxygen, at a distance from the phosphorus atom of the α-phosphate (3.69 Å) consistent with and supporting a catalytic mechanism involving two divalent metal ions. Finally, soaking with MnCl 2 converted a pre-catalytic Pol λ/Na + complex with unreacted dCTP in the active site into a product complex via catalysis in the crystal. These data provide pre-and post-transition state information and outline in a single crystal the pathway for the phosphoryl transfer reaction carried out by DNA polymerases.

show abstract

“…Pol k acts as a backup for Pol b function in general BER [Braithwaite et al, 2005;Tano et al, 2007;Braithwaite et al, 2010], but has a specific role in BER of the oxidized base 8-oxo-deoxyguanine (8-oxoG). Together with cofactors replication protein A and proliferating cell nuclear antigen, Pol k is the mammalian polymerase most able to correctly incorporate dC opposite 8-oxoG [Maga et al, 2007].…”

Section: Pol Kmentioning

confidence: 99%

DNA polymerases in nonhomologous end joining: Are there any benefits to standing out from the crowd?

Ramsden

Asagoshi

2012

Environ and Mol Mutagen

View full text Add to dashboard Cite

Chromosome breaks, often with damaged or missing DNA flanking the break site, are an important threat to genome stability. They are repaired in vertebrates primarily by nonhomologous end joining (NHEJ). NHEJ is unique among the major DNA repair pathways in that a continuous template cannot be used by DNA polymerases to instruct replacement of damaged or lost DNA. Nevertheless, at least 3 out of the 17 mammalian DNA polymerases are specifically employed by NHEJ. Biochemical and structural studies are further revealing how each of the polymerases employed by NHEJ possesses distinct and sophisticated means to overcome the barriers this pathway presents to polymerase activity. Still unclear, though, is how the resulting network of overlapping and nonoverlapping polymerase activities contributes to repair in cells. Environ. Mol. Mutagen. 53:741-751, 2012. V V C 2012 Wiley Periodicals, Inc.

show abstract

DNA Polymerase λ Mediates a Back-up Base Excision Repair Activity in Extracts of Mouse Embryonic Fibroblasts

Cited by 146 publications

References 39 publications

DNA polymerase beta null mouse embryonic fibroblasts harbor a homozygous null mutation in DNA polymerase iota

DNA polymerase beta null mouse embryonic fibroblasts harbor a homozygous null mutation in DNA polymerase iota

Role of the catalytic metal during polymerization by DNA polymerase lambda

DNA polymerases in nonhomologous end joining: Are there any benefits to standing out from the crowd?

Contact Info

Product

Resources

About