Cyclobutane thymine dimers in a<i>ras</i>proto-oncogene hot spot activate the gene by point mutation

Kamiya, Hiroyuki; Murata, Naho; Murata, Tetsuya; Iwai, Shigenori; Matsukage, Akio; Masutani, Chikahide; Hanaoka, Fumio; Ohtsuka, Eiko

doi:10.1093/nar/21.10.2355

Cited by 19 publications

(19 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…DNA polymerase 1 has been reported to copy past apurinic sites (27) and to synthesize in vitro through a cis-syn thymine dimer placed on codon 61 of HRAS gene (28); however, in contrast to what we observed with the Pt-d(GpG) adduct, the translesion replication of a cis-syn dimer does not appear to be specific to DNA polymerase 13, since it is also bypassed by DNA polymerase a (28) and DNA polymerase 6 in the presence of PCNA (23). It is possible that translesion synthesis observed here may be influenced by the type of lesion and the sequence context.…”

Section: Discussionmentioning

confidence: 99%

DNA polymerase beta bypasses in vitro a single d(GpG)-cisplatin adduct placed on codon 13 of the HRAS gene.

Hoffmann

Pillaire

Maga

et al. 1995

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

117

108

View full text Add to dashboard Cite

We have examined the capacity of calf thymus DNA polymerases a, f, 6, and e to perform in vitro translesion synthesis on a substrate containing a single d(GpG)-cisplatin adduct placed on codon 13 of the human HRAS gene. We found that DNA synthesis catalyzed by DNA polymerases ca, 6, and E was blocked at the base preceding the lesion Among the possible mechanisms of mutagenesis is error-prone replication by cellular DNA polymerases past a DNA lesion followed by fixation of the mutation during subsequent rounds of replication. In eukaryotic cells, it is still unclear whether the replicative DNA polymerases a, 8, and E can carry out translesion synthesis either alone or with the help of accessory proteins. Alternatively, a separate DNA polymerase may be required for this process. For example, genetic evidence in the yeast Saccharomyces cerevisiae indicates that a putative DNA polymerase, the product of the REV3 gene, may be involved in error-prone translesion synthesis but not in normal DNA replication (1). DNA polymerase (3 is one of the five mammalian polymerases identified to date and is believed to function primarily in the repair of damaged DNA (2). However, DNA polymerase a may also have a role in replicative synthesis, since the enzyme can substitute for DNA polymerase I during DNA replication in Escherichia coli (3), and it is required for the conversion of single-stranded M13 DNA to double-stranded DNA in Xenopus oocytes and in oocyte nuclear extracts (4). cis-Diamminedichloroplatinum(II) (cisplatin) is an anticancer agent widely used in the treatment of ovarian, testicular, head, and neck carcinomas (5). It is believed that this compound exerts its cytotoxic properties by forming stable lesions on DNA, primarily intrastrand cross-links at the N-7 positions of adjacent guanine bases [d(GpG)-cisplatin or Pt-d(GpG)] (6). Replicative bypass of cisplatin adducts has been described in bacteria (7,8) and in eukaryotic cells (9). Recent work in our laboratory has demonstrated that a single-stranded DNA The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.vector bearing a unique intrastrand bifunctional adduct Ptd(GpG) at codon 13 of the human protooncogene HRAS is replicated in simian COS-7 cells and that such translesion synthesis may be mutagenic (10).To our knowledge, the capacities of the major mammalian replication enzymes to bypass the Pt-d(GpG) lesion have not been compared. To address this question, we have investigated the ability of purified calf thymus DNA polymerases a, (3, 8, and s to catalyze in vitro the bypass synthesis of a single Pt-d(GpG) adduct placed on codon 13 of the human HRAS protooncogene, the same sequence used for our previous in vivo studies (10). Results show that only DNA polymerase 3 is capable of in vitro translesional synthesis and indicate that its ability to initiate DNA replication opposite the cisplatin adduct may ...

show abstract

Section: Discussionmentioning

confidence: 99%

DNA polymerase beta bypasses in vitro a single d(GpG)-cisplatin adduct placed on codon 13 of the HRAS gene.

Hoffmann

Pillaire

Maga

et al. 1995

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

117

108

View full text Add to dashboard Cite

show abstract

“…Construction of the c-Ha-ras gene with OH8Ade and transfection of the gene into NIH 3T3 cells Preparation of the DNA cassette and insertion into the vector was done as described previously (26,33). The vector thus obtained (100, 150 and 500 ng), with 30 jig genomic DNA isolated from NIH 3T3 cells, was transfected into NIH 3T3 cells by the calcium phosphate procedure, as described (34,35).…”

Section: Dna Polymerase Reactions and Analysis Of Incorporated Nucleomentioning

confidence: 99%

“…The vector thus obtained (100, 150 and 500 ng), with 30 jig genomic DNA isolated from NIH 3T3 cells, was transfected into NIH 3T3 cells by the calcium phosphate procedure, as described (34,35). GAG ACC TGT CTG CTG GAT ATC CTT GAT ACC GCA GGC C*A GAA GAA TAC TCT GCG ATG CGT GAT CAG TAT ATG CGT ACC 3' 3' GC TAC GCA CTA GTC ATA TAC GCA TGG CCG CTT CCG AAG 5 Analysis of c-Ha-ras genes present in transformed cells c-Ha-ras genes present in transformed cells were analyzed by the PCR-RE method described previously (8,26). Briefly, the ras gene fragments surrounding codon 61 (204 bp) were amplified by the polymerase chain reaction (PCR) (36) using U5 (5'-dGAAG-ACTCTTACCGTAAGC-3', which corresponds to the region from codon 37 to 43 of the ras gene) and L9 (5'-dTllTAACGCG-lITITGATCTGTFCACGGTATTGATGGATGTC-TTC-3', which corresponds to the region from codon 91 to 104) (31) as primers and the PCR-RE method was carried out.…”

Section: Dna Polymerase Reactions and Analysis Of Incorporated Nucleomentioning

confidence: 99%

“…An abasic site and its analog induce a variety of mutations in mammalian cells (18)(19)(20), while they elicit the incorporation of dAMP opposite their positions in vitro (18,(21)(22)(23) and mutations to T in bacteria (24,25). We previously demonstrated that cis-syn and trans-syn cyclobutane thymine dimers in the same nucleotide sequences showed different mutagenic potentials and mutation spectra in in vitro and mammalian systems (26). Moreover, nucleotide incorporation opposite ethenocytosine and propanoguanine is markedly different in bacterial and mammalian cells (27 induce mutations in mammalian cells, although the mutagenicity of the modifled base is negligible in Escherichia coli (17).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

8-Hydroxyadenine (7, 8-dihydro-8-Oxoadenine) induces misincorporation inin vitroDNA synthesis and mutations in NIH 3T3 cells

Kamiya¹,

Miura²,

Murata‐Kamiya³

et al. 1995

Nucl Acids Res

Self Cite

132

View full text Add to dashboard Cite

An oligodeoxyribonucleotide containing 8-hydroxyadenine (OH8Ade) was chemically synthesized and single- and double-stranded c-Ha-ras gene fragments with OH8Ade at the second position of codon 61 were prepared. The single-stranded ras gene fragment was used as a template for in vitro DNA synthesis with the Klenow fragment of Escherichia coli DNA polymerase I, Taq DNA polymerase, rat DNA polymerase beta and mouse DNA polymerase alpha. The former two enzymes exclusively incorporated dTMP opposite OH8Ade. The DNA polymerases alpha and beta misinserted dGMP, and dAMP and dGMP, respectively. The c-Ha-ras gene was constructed using the double-stranded ras gene fragment containing OH8Ade and was transfected into NIH 3T3 cells. The gene with OH8Ade induced focus formation, indicating that OH8Ade elicited point mutations in cells. When c-Ha-ras genes present in transformed cells were analyzed, an A-->G transition and an A-->C transversion were detected. These results indicate that OH8Ade induced misincorporation in in vitro DNA synthesis and mutations in mammalian cells.

show abstract

“…8) Organic syntheses in combination with joining enzyme systems provide various approaches to studies of gene functions by preparation of genes that contain a modified base. Modified bases, O 6 -methylguanine, 9) 7, 8-dihydro-8-oxoguanine (8-oxoguanine), 10) pyrimidine photo-dimers 11) have been included by using chemically synthesized oligonucleotides. One of the oxidative products of guanine, 8-oxoguanine, was found to be formed by oxygen or X-ray radiation (Fig.…”

mentioning

confidence: 99%

Non-canonical hydrogen bonds in genetic information flow

Ohtsuka

2005

Proceedings of the Japan Academy. Ser. B: Physical and Biologic

Self Cite

View full text Add to dashboard Cite

Non-Watson-Crick-type hydrogen bonds involving minor nucleosides such as inosine in anticodon loops of transfer RNA are found in nucleic acids. Inosine contains hypoxanthine as a nucleobase and can form base pairs with various bases in the genetic decoding process. This property was applied to cloning complementary DNA by using oligodeoxyribonucleotide primers which contain deoxyinosine residues. Hypoxanthine is generated by deamination of adenine, one of the major nucleobases in DNA and RNA. Formation of unusual hydrogen bonds derived from damaged nucleobases in nucleic acids is thought to be main cause of disorders in genetic information flow. Mutagenesis and carcinogenesis of damaged bases, including hypoxanthine, 7, 8-dihydro-8-oxoguanine (8-oxoguanine) and pyrimidine photo-dimers in c-Haras genes, were investigated by using synthetic genes containing modified nucleotides. The mode of recognition of unusual base pairing between a thymine photo-dimer and adenine by a repair enzyme was studied by X-ray analysis.

show abstract

Cyclobutane thymine dimers in arasproto-oncogene hot spot activate the gene by point mutation

Cited by 19 publications

References 44 publications

DNA polymerase beta bypasses in vitro a single d(GpG)-cisplatin adduct placed on codon 13 of the HRAS gene.

DNA polymerase beta bypasses in vitro a single d(GpG)-cisplatin adduct placed on codon 13 of the HRAS gene.

8-Hydroxyadenine (7, 8-dihydro-8-Oxoadenine) induces misincorporation inin vitroDNA synthesis and mutations in NIH 3T3 cells

Non-canonical hydrogen bonds in genetic information flow

Contact Info

Product

Resources

About