Immortalized mouse cell lines that lack a functional Rev3 gene are hypersensitive to UV irradiation and cisplatin treatment

Zander, Linda; Bemark, Mats

doi:10.1016/j.dnarep.2004.03.031

Cited by 38 publications

(30 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The important role of pol in this process is consistent with the situation in S. cerevisiae (28), and with its essential role during mouse embryonic development (5,17). It is also consistent with the increased UV sensitivity of normal, XPV and XPA cells in which the expression of REV3L was knocked-down, as shown in this study, and in other cell types in previous studies (15,(29)(30)(31). There are also earlier studies, which reported no effect of REV3L on UV sensitivity; this discrepancy might be due to differences in cell lines and methodologies (e.g., antisense RNA versus siRNA) (32)(33)(34).…”

Section: Discussionsupporting

confidence: 91%

DNA polymerase ζ cooperates with polymerases κ and ι in translesion DNA synthesis across pyrimidine photodimers in cells from XPV patients

Ziv

Geacintov

Nakajima

et al. 2009

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

114

125

View full text Add to dashboard Cite

Human cells tolerate UV-induced cyclobutane pyrimidine dimers (CPD) by translesion DNA synthesis (TLS), carried out by DNA polymerase , the POLH gene product. A deficiency in DNA polymerase due to germ-line mutations in POLH causes the hereditary disease xeroderma pigmentosum variant (XPV), which is characterized by sunlight sensitivity and extreme predisposition to sunlight-induced skin cancer. XPV cells are UV hypermutable due to the activity of mutagenic TLS across CPD, which explains the cancer predisposition of the patients. However, the identity of the backup polymerase that carries out this mutagenic TLS was unclear. Here, we show that DNA polymerase cooperates with DNA polymerases and to carry out error-prone TLS across a TT CPD. Moreover, DNA polymerases and , but not , protect XPV cells against UV cytotoxicity, independently of nucleotide excision repair. This presents an extreme example of benefit-risk balance in the activity of TLS polymerases, which provide protection against UV cytotoxicity at the cost of increased mutagenic load.carcinogenesis ͉ DNA repair ͉ lesion bypass ͉ replication ͉ ultraviolet T LS is a fundamental mechanism for tolerating DNA damage that has escaped repair, carried out by specialized lowfidelity DNA polymerases, which synthesize across a wide variety of DNA lesions (1). At least 5 TLS DNA polymerases are present in mammals, four of which, DNA polymerases , , , and REV1, belong to the Y superfamily. The fifth TLS polymerase is pol, which belongs to the B family, and is the only TLS polymerase known to be essential in mammals (2-5). TLS polymerases exhibit a certain degree of specificity for their substrate DNA lesions, and their activity is tightly regulated (6-9). The most well characterized TLS polymerase is pol, which is specialized to bypass cyclobutane pyrimidine dimers (CPD) in a relatively error-free manner. The biological significance of pol is illustrated by the hereditary disease xeroderma pigmentosum variant (XPV), in which germ-line mutations in the POLH gene, encoding pol, cause an extreme 1000-fold increased predisposition to sunlight-induced skin cancer (10, 11). Cells from XPV patients exhibit a slightly increased UV sensitivity, and a dramatic UV hypermutability (12), which is responsible for their extreme cancer predisposition. The UV hypermutability is explained by the activity of a back-up DNA polymerase that performs TLS across CPD with lower efficiency and higher error-frequency. Although there is evidence that pol is involved in TLS across CPD in XPV cells (13-15), additional polymerases may be involved, and the picture is far from being complete. This is an important issue because these polymerases are likely to be driving sunlight-induced skin carcinogenesis in XPV patients.Here, we show that 3 TLS polymerases, pol, pol, and pol, are involved in TLS across CPD in XPV cells. Moreover, pol and pol, but not pol, also provide protection against UV cytotoxicity, independently of nucleotide excision repair (NER). Results Pol, pol, and pol Are Involved in T...

show abstract

Section: Discussionsupporting

confidence: 91%

DNA polymerase ζ cooperates with polymerases κ and ι in translesion DNA synthesis across pyrimidine photodimers in cells from XPV patients

Ziv

Geacintov

Nakajima

et al. 2009

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

114

125

View full text Add to dashboard Cite

show abstract

“…Most likely, a p53-dependent checkpoint is only partly responsible for the arrested division observed in cultured Rev3L À/À cells. A similar conclusion was reached by Zander and Bemark, who also generated proliferating Rev3L null MEFs in a p53 mutant background (44). The population doubling time of our Rev3L À/À embryonic fibroblasts was f11% longer than a Rev3L +/+ control, whereas the plating efficiency and saturation density were higher for the Rev3L null cells.…”

Section: Mitotic Activity Of Rev3lsupporting

confidence: 87%

“…DT40 cells deleted for Rev3 are sensitive to these four types of DNA damage, with a notable difference being the pronounced sensitivity of these cells to cisplatin (45). Independently isolated Rev3L À/À ;p53 À/À MEFs are modestly sensitive to both cisplatin and UV radiation (44). A REV3L antisense knockdown cell line, which retains a low level of REV3L expression, is mildly sensitive to cisplatin and has little or no UV sensitivity (21,54).…”

Section: Cancer Researchmentioning

confidence: 99%

Loss of DNA Polymerase ζ Causes Chromosomal Instability in Mammalian Cells

et al. 2006

View full text Add to dashboard Cite

Rev3L encodes the catalytic subunit of DNA polymerase Z (pol Z) in mammalian cells. In yeast, pol Z helps cells bypass sites of DNA damage that can block replication enzymes. Targeted disruption of the mouse Rev3L gene causes lethality midway through embryonic gestation, and Rev3L À/À mouse embryonic fibroblasts (MEFs) remain in a quiescent state in culture. This suggests that pol Z may be necessary for tolerance of endogenous DNA damage during normal cell growth. We report the generation of mitotically active Rev3L À/À MEFs on a p53 À/À genetic background.Rev3L null MEFs exhibited striking chromosomal instability, with a large increase in translocation frequency. Many complex genetic aberrations were found only in Rev3L null cells. Rev3L null cells had increased chromosome numbers, most commonly near pentaploid, and double minute chromosomes were frequently found. This chromosomal instability associated with loss of a DNA polymerase activity in mammalian cells is similar to the instability associated with loss of homologous recombination capacity. Rev3L null MEFs were also moderately sensitive to mitomycin C, methyl methanesulfonate, and UV and ;-radiation, indicating that mammalian pol Z helps cells tolerate diverse types of DNA damage. The increased occurrence of chromosomal translocations in Rev3L À/À MEFs suggests that loss of Rev3L expression could contribute to genome instability during neoplastic transformation and progression. (Cancer Res 2006; 66(1): 134-42)

show abstract

“…After the knockdown of hRev3 expression, both cell lines had 2-to 3-fold increased sensitivity when compared to the control cells. This is consistent with the finding that Rev3 knockout mouse fibroblast cells showed an increase in cisplatin sensitivity (10). Thus, Polζ plays an important role in DNA repair, which in turn plays a role in resistance to cisplatin.…”

Section: Discussionsupporting

confidence: 92%

Hypersensitivity to cisplatin after hRev3 mRNA knockdown in head and neck squamous cell carcinoma cells

Adachi

Ijichi²,

Hasegawa³

et al. 2008

Mol Med Rep

View full text Add to dashboard Cite

Abstract. Resistance to platinum-based chemotherapy frequently poses a significant problem in the treatment of head and neck squamous cell carcinomas (HNSCCs). In this study, we isolated cisplatin-resistant UM-SCC-23 CDDP/R cells from a UM-SCC-23 head and neck squamous cell carcinoma cell line. The UM-SCC-23 CDDP/R cells were approximately 3.5-fold more resistant to cisplatin than the UM-SCC-23 cells. Translesion DNA synthesis (TLS) is one of the major pathways involved in post-replication repair. To ascertain whether TLS is involved in cisplatin resistance in human cancer cells, we analyzed expression of the Rev3 gene, which encodes the catalytic subunit of DNA polymerase ζ (Polζ), using quantitative PCR. Expression of hRev3 mRNA in the UM-SCC-23 CDDP/R cells was approximately 1.4-fold higher than in the UM-SCC-23 cells. In both types of cells, expression of hRev3 mRNA was suppressed using siRNAs. Cisplatin sensitivity after hRev3 mRNA knockdown increased approximately 2-fold in UM-SCC-23 cells and approximately 3-fold in UM-SCC-23 CDDP/R cells. This study suggests that hRev3 knockdown with siRNAs increases sensitivity to cisplatin. Inhibition of the hRev3 gene may therefore prove to be a novel clinical strategy for reducing resistance to platinumbased chemotherapy in HNSCC. IntroductionCisplatin-based chemotherapy is widely used in the management of locally-advanced head and neck squamous cell carcinoma (HNSCC). However, recurrence often occurs following primary treatment, and treatment of recurrent tumors frequently fails due to resistance to therapeutic agents, including cisplatin (1). Although, to date, there have been many studies concerning the mechanisms of chemoresistance, effective strategies against chemoresistance in HNSCC (2-4) have yet to be devised.The major cytotoxic effect of cisplatin is through the formation of DNA damage, such as DNA-interstrand crosslinks (4). After exposure to cisplatin, cells show one of two reactions: the induction of apoptosis through signal transduction, trigged by DNA damage and resulting in cell death, or the repair of DNA damage for cell survival (5).Translesion DNA synthesis (TLS) and homologous recombination (HR) are the two major pathways involved in post replication repair (6). A number of TLS polymerases have been identified in Saccharomyces cerevisiae and mammals (7).Candidates for translesion synthesis past bulky DNA adducts in vivo include members of the DNA polymerase X (polymerase ß, polymerase μ, polymerase λ), polymerase B (polymerase ζ) and polymerase Y (Rev1, polymerase η, polymerase τ, polymerase κ) families. Of these, DNA polymerase η exhibits the highest translesion activity for the cisplatin adduct, followed by polymerase μ and polymerase ζ (8).The Rev3 and Rev7 genes encode the catalytic subunit of DNA polymerase ζ (Polζ) (7). Inactivation of the Rev3 gene sensitized chicken cells to cisplatin and to various DNA damaging agents, such as UV, methyl methanesulfonate and ionizing radiation. hRev3 antisense-expressing human fibroblasts demonstrated hyp...

show abstract

Immortalized mouse cell lines that lack a functional Rev3 gene are hypersensitive to UV irradiation and cisplatin treatment

Cited by 38 publications

References 32 publications

DNA polymerase ζ cooperates with polymerases κ and ι in translesion DNA synthesis across pyrimidine photodimers in cells from XPV patients

DNA polymerase ζ cooperates with polymerases κ and ι in translesion DNA synthesis across pyrimidine photodimers in cells from XPV patients

Loss of DNA Polymerase ζ Causes Chromosomal Instability in Mammalian Cells

Hypersensitivity to cisplatin after hRev3 mRNA knockdown in head and neck squamous cell carcinoma cells

Contact Info

Product

Resources

About