UV stimulation of DNA-mediated transformation of human cells.

Duin, M van; Westerveld, A; Hoeijmakers, Jan H.J.

doi:10.1128/mcb.5.4.734

Cited by 47 publications

(16 citation statements)

References 36 publications

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“…No UV-dependent enhancement of plasmid-chromosome recombination was observed, in contrast to reports that homologous interviral recombination (13), homologous intrachromosomal recombination (72), and nonhomologous plasmid integration (43,59,65) were enhanced by UV irradiation. Gamma irradiation and double-strand breaks enhanced pSV3neo-Ap-recombination, so the observation that UV light increased the frequency of mutant (mostly degraded) plasmids almost 10-fold but did not affect recombination was a bit contradictory.…”

Section: Discussioncontrasting

confidence: 49%

Recombination between irradiated shuttle vector DNA and chromosomal DNA in African green monkey kidney cells.

Mudgett¹,

Taylor²

1990

Mol. Cell. Biol.

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An autonomously replicating shuttle vector was used to investigate enhancement of plasmid-chromosome recombination in mammalian host cells by gamma irradiation and UV light. Sequences homologous to the shuttle vector were stably inserted into the genome of African green monkey kidney cells to act as the target substrate for these recombination events. The shuttle vector molecules were irradiated at various doses before transfection into the mammalian host cells that contained the stable insertions. The homologous transfer of the bacterial ampicillin resistance gene from the inserted sequences to replace a mutant ampicillin sensitivity gene on the shuttle vector was identified by the recovery of ampicillin-resistant plasmids after Hirt extraction and transformation into Escherichia coli host cells. Gamma irradiation increased homologous shuttle vector-chromosome recombination, whereas UV light did not increase the frequency of recombinant plasmids detected. Introducing specific double-strand breaks in the plasmid or prolonging the time of plasmid residence in the mammalian host cells also enhanced plasmid-chromosome recombination. In contrast, plasmid mutagenesis was increased by UV irradiation of the plasmid but did not change with time. The ampicillin-resistant recombinant plasmid molecules analyzed appeared to rise mostly from nonconservative exchanges that involved both homologous and possibly nonhomologous interactions with the host chromosome. The observation that similar recombinant structures were obtained from all the plasmid treatments and host cells used suggests a common mechanism for plasmid-chromosome recombination in these mammalian cells.

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Section: Discussioncontrasting

confidence: 49%

Recombination between irradiated shuttle vector DNA and chromosomal DNA in African green monkey kidney cells.

Mudgett¹,

Taylor²

1990

Mol. Cell. Biol.

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“…We and others have demonstrated that cyclobutane pyrimidine dimers are predominantly responsible for shortwavelength UV-enhanced transformation (36,42), whereas DNA lesions such as thymine glycols or apurinic sites do not induce the phenomenon (36). Stimulation of transformation with UV-induced pyrimidine dimers and with psoralen adducts both involve lesions in cyclobutane rings on pyrimidine bases.…”

Section: Discussionmentioning

confidence: 99%

“…As a model system to study chromosomal rearrangements, we are using integration of plasmid DNA into mammalian chromosomes. We and others have demonstrated that UV irradiation of pSV2-derived shuttle vectors enhances the potential of the vectors to transform human cells (35,42). Integration of the vector DNA into host chromosomes appears to occur through nonhomologous recombination (20,34,37,39).…”

mentioning

confidence: 99%

DNA interstrand cross-links promote chromosomal integration of a selected gene in human cells.

Vos¹,

Hanawalt²

1989

Mol. Cell. Biol.

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We have used integrative pSV2 plasmids to learn how DNA lesions affect nonhomologous recombination with human chromosomes. Enhanced stable transformation of fibrosarcoma cells with a selectable gene was observed after chemical modification of the plasmid DNA; thus, cells transfected with plasmid pSV2-gpt carrying photoadducts of the cross-linking agent 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) yielded four-to sevenfold-higher levels of Gpt+ transformants than were obtained with untreated plasmid. The enhancement due to HMT interstrand cross-links was at least as great as that due to the monoadducts. DNA hybridization analysis indicated that the enhanced transformation frequency resulted from an increased number of cells carrying integrated plasmid sequences rather than from a higher copy number per transformant. The enhancement was not seen with a plasmid missing the sequences flanking the minimal simian virus 40 gpt transcription unit. Cotransfection with untreated and HMT-treated plasmids suggested that the HMT-containing DNA interacted preferentially with some cellular factor that promoted chromosomal integration of the plasmid DNA. It is concluded that (i) interstrand cross-linking as well as intrastrand DNA adducts promote nonhomologous recombination in human chromatin and (ii) DNA sequences flanking the selectable genes are the targets for such recombinational events.

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“…To date, the use of defective human cell lines such as XP, Fanconi's anemia, and ataxia telangiectasia in transformation experiments with genomic DNAs has been unsuccessful (20). This is due, at least in part, to the reduced amounts of stably integrated DNA in human lines relative to many rodent lines (14,47). Some success with correction of XP cells has been achieved by fusion of X-ray-inactivated CHO cells with XP group A (17), resulting in primary hybrids having -2% hamster DNA.…”

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confidence: 99%

Molecular cloning and biological characterization of a human gene, ERCC2, that corrects the nucleotide excision repair defect in CHO UV5 cells.

et al. 1988

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The UV-sensitive Chinese hamster ovary (CHO) cell line UV5, which is defective in the incision step of nucleotide excision repair, was used to identify and clone a complementing human gene, ERCC2, and to study the repair process. Genomic DNA from a human-hamster hybrid cell line was sheared and cotransferred with pSV2gpt plasmid DNA into UV5 ceUs to obtain five primary transformants. Transfer of sheared DNA from one primary transformant resulted in a secondary transformant expressing both gpt and ERCC2. The human repair gene was identified with a probe for Alu-family repetitive sequences. For most primary, secondary, and cosmid transformants, survival after UV exposure showed a return to wild-type levels of resistance. The levels of UV-induced mutation at the aprt locus for secondary and cosmid transformants varied from 50 to 130% of the wild-type level. Measurements of the initial rate of UV-induced strand incision by alkaine elution indicated that, whereas the UV5 rate was 3% of the wild-type level, rates of cosmid-transformed lines were similar to that of the wild type, and the secondary transformant rate was about 165% of the wild-type rate. Analysis of overlapping cosmids determined that ERCC2 is between 15.5 and 20 kilobases and identified a closely linked gpt gene. Cosmids were obtained with functional copies of both ERCC2 and gpt. ERCC2 corrects only the first of the five CHO complementation groups of incision-defective mutants.

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UV stimulation of DNA-mediated transformation of human cells.

Cited by 47 publications

References 36 publications

Recombination between irradiated shuttle vector DNA and chromosomal DNA in African green monkey kidney cells.

Recombination between irradiated shuttle vector DNA and chromosomal DNA in African green monkey kidney cells.

DNA interstrand cross-links promote chromosomal integration of a selected gene in human cells.

Molecular cloning and biological characterization of a human gene, ERCC2, that corrects the nucleotide excision repair defect in CHO UV5 cells.

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