DNA strand specificity for UV-induced mutations in mammalian cells.

Vrieling, Harry; Rooijen, M L Van; Groen, Nicole A.; Zdzienicka, M. Z.; Simons, J.W.I.M.; Lohman, P.H.M.; Zeeland, A.A. van

doi:10.1128/mcb.9.3.1277

Cited by 157 publications

(65 citation statements)

References 30 publications

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“…Another example of higher mutagenesis in the TS was observed previously in mfd-deficient E. coli lacking the TCR process (Oller et al 1992). These results are in contrast with those of the TSs of RNAP II genes that are preferentially repaired and, consequently, show a lower mutagenesis as compared with the NTSs (Vrieling et al 1989;McGregor et al 1991;Sage et al 1993). In conclusion, these data establish a direct link between NER and mutagenesis, and indicate that RNAP III transcription is not coupled to NER and that, in opposition to RNAP II genes, it is the NTSs of RNAP III genes that are preferentially repaired.…”

Section: Mutagenesis Is Linked To Nucleotide Excision Repaircontrasting

confidence: 54%

Nucleotide excision repair and photolyase preferentially repair the nontranscribed strand of RNA polymerase III-transcribed genes in Saccharomyces cerevisiae

Aboussekhra¹,

Thoma²

1998

Genes & Development

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A high-resolution primer extension technique was used to study the relationships between repair, transcription, and mutagenesis in RNA polymerase III transcribed genes in Saccharomyces cerevisiae. The in vivo repair of UV-induced DNA damage by nucleotide excision repair (NER) and by photoreactivation is shown to be preferential for the nontranscribed strand (NTS) of the SNR6 gene. This is in contrast to RNA polymerase II genes in which the NER is preferential for the transcribed strand (TS). The repair-strand bias observed in SNR6 was abolished by inactivation of transcription in a snr6⌬2 mutant, showing a contribution of RNA polymerase III transcription in this phenomenon. The same strand bias for NER (slow in TS, fast in NTS) was discovered in the SUP4 gene, but only outside of the intragenic promoter element (box A). Unexpectedly, the repair in the transcribed box A was similar on both strands. The strand specificity as well as the repair heterogeneity determined in the transcribed strand of the SUP4 gene, correlate well with the previously reported site-and strand-specific mutagenesis in this gene. These findings present a novel view regarding the relationships between DNA repair, mutagenesis, and transcription.

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Section: Mutagenesis Is Linked To Nucleotide Excision Repaircontrasting

confidence: 54%

Nucleotide excision repair and photolyase preferentially repair the nontranscribed strand of RNA polymerase III-transcribed genes in Saccharomyces cerevisiae

Aboussekhra¹,

Thoma²

1998

Genes & Development

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“…But evidence does exist linking mismatch repair activities with TCR (4,17,22); thus, the factors regulating the process of gene repair remain to be fully defined. For example, an elevated number of mutations have been observed to form initially on the nontranscribed strand of the human HPRT gene, a gene transcribed by RNA polymerase II (34,35). Thus, enzymatic processing events during transcription can actually be focused toward the nontranscribed strand.…”

Section: Discussionmentioning

confidence: 99%

Strand Bias in Targeted Gene Repair Is Influenced by Transcriptional Activity

Liu

Rice

Drury

et al. 2002

Molecular and Cellular Biology

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Modified single-stranded DNA oligonucleotides can direct nucleotide exchange in Saccharomyces cerevisiae. Point and frameshift mutations are corrected in a reaction catalyzed by cellular enzymes involved in various DNA repair processes. The present model centers on the annealing of the vector to one strand of the helix, followed by the correction of the designated base. The choice of which strand to target is a reaction parameter that can be controlled, so here we investigate the properties of strand bias in targeted gene repair. An in vivo system has been established in which a plasmid containing an actively transcribed, but mutated, hygromycinenhanced green fluorescent protein fusion gene is targeted for repair and upon conversion will confer hygromycin resistance on the cell. Overall transcriptional activity has a positive influence on the reaction, elevating the frequency. If the targeting vector is synthesized so that it directs nucleotide repair on the nontranscribed strand, the level of gene repair is higher than if the template strand is targeted. We provide data showing that the targeting vector can be displaced from the template strand by an active T7 phage RNA polymerase. The strand bias is not influenced by which strand serves as the leading or lagging strand during DNA synthesis. These results may provide an explanation for the enhancement of gene repair observed when the nontemplate strand is targeted.The use of oligonucleotides to direct single-base changes in DNA has become an important strategy for understanding gene function. While processes for disabling targeted genes and creating knockout strains have provided an enormous amount of information, it is now becoming evident that subtle changes in the genomes of many organisms constitute a centerpiece of genetic variability. Specifically, as information regarding the human genome accumulates, the importance of single-nucleotide polymorphisms is now clear. These simple alterations, often causing no visible phenotype variation, may hold the key to the spectrum of responses to many therapeutic agents. In recognition of the importance of single-nucleotide polymorphisms, several technologies have emerged that aim to create animal and plant models that will recapitulate the nucleotide alterations without disrupting the integrity of the whole gene. Many of these technologies employ synthetic oligonucleotides as vectors for directing the introduction of the specific changes.Our laboratory has developed a series of these oligonucleotide vectors that can create single-base changes in chromosomal and episomal targets. One of these vectors is the chimeric RNA/DNA oligonucleotide (chimera), which consists of cRNA and DNA residues folded into a double hairpin conformation. Chimeras have been shown to direct base replacement or base insertion in yeast (21, 27), mammalian cells in culture (1, 6, 15, 38), animal models (2, 14, 16, 26, 31), and plants (3,28,40). While the mechanism of targeted gene repair is not fully elucidated, a number of important steps have...

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“…In general, the e ects of transcription on repair are more pronounced at low levels of DNA damage (Vrieling et al, 1989(Vrieling et al, , 1991. Since much higher levels of DNA damage are typically employed for repair assays than for mutation assays, this may be the reason that the strand bias of mutation is much more pronounced than the strand bias of repair.…”

Section: Discussionmentioning

confidence: 99%

Slow repair of bulky DNA adducts along the nontranscribed strand of the human p53 gene may explain the strand bias of transversion mutations in cancers

et al. 1998

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Using UvrABC incision in combination with ligationmediated PCR (LMPCR) we have previously shown that benzo(a)pyrene diol epoxide (BPDE) adduct formation along the nontranscribed strand of the human p53 gene is highly selective; the preferential binding sites coincide with the major mutation hotspots found in human lung cancers. Both sequence-dependent adduct formation and repair may contribute to these mutation hotspots in tumor tissues. To test this possibility, we have extended our previous studies by mapping the BPDE adduct distribution in the transcribed strand of the p53 gene and quantifying the rates of repair for individual damaged bases in exons 5, 7, and 8 for both DNA strands of this gene in normal human ®broblasts. We found that: (i) on both strands, BPDE adducts preferentially form at CpG sequences, and (ii) repair of BPDE adducts in the transcribed DNA strand is consistently faster than repair of adducts in the nontranscribed strand, while repair at the major damage hotspots (guanines at codons 157, 248 and 273) in the nontranscribed strand is two to four times slower than repair at other damage sites. These results strongly suggest that both preferential adduct formation and slow repair lead to hotspots for mutations at codons 157, 248 and 273, and that the strand bias of bulky adduct repair is primarily responsible for the strand bias of G to T transversion mutations observed in the p53 gene in human cancers.

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DNA strand specificity for UV-induced mutations in mammalian cells.

Cited by 157 publications

References 30 publications

Nucleotide excision repair and photolyase preferentially repair the nontranscribed strand of RNA polymerase III-transcribed genes in Saccharomyces cerevisiae

Nucleotide excision repair and photolyase preferentially repair the nontranscribed strand of RNA polymerase III-transcribed genes in Saccharomyces cerevisiae

Strand Bias in Targeted Gene Repair Is Influenced by Transcriptional Activity

Slow repair of bulky DNA adducts along the nontranscribed strand of the human p53 gene may explain the strand bias of transversion mutations in cancers

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