Isolation of Mutants of an Animal Virus in Bacteria

Peden, Keith; Pipas, James M.; Pearson‐White, Sonia; Nathans, Daniel

doi:10.1126/science.6251547

Cited by 343 publications

(198 citation statements)

References 35 publications

(15 reference statements)

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“…All replication assays in this study used the standard procedure of measuring DpnI-resistant DNA (34). The input bacterial plasmid DNA contains sites susceptible to DpnI cleavage which are converted to DpnI-resistant sites when DNA replicationdependent methylation changes occur in the COS cells.…”

Section: Resultsmentioning

confidence: 99%

The replication activation potential of selected RNA polymerase II promoter elements at the simian virus 40 origin.

Hoang¹,

Wang²,

Gralla³

1992

Mol. Cell. Biol.

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Binding sites for cellular transcription factors were placed near the simian virus 40 origin of replication, and their effect on replication and TATA-dependent transcription was measured in COS cells. The hierarchy of transcriptional stimulation changed when the plasmids replicated. Only one of seven inserted sequences, a moderately weak transcription element, stimulated replication detectably. However, when two nonstimulatory sites were present in multiple copies they did activate replication. Multiple sites for the chimeric activator GAL4-VP16 did not stimulate replication even though transcription was stimulated strongly. The results indicate that the ability of a binding site to stimulate replication from the simian virus 40 ori is not based on its transcriptional activation potential but is instead related to a separate replication activation potential that can be increased by having multiple sites.The interrelationship between control of cellular replication and transcription has been the subject of extensive experimentation and discussion (10,11,19,38). Because of the uncertainty concerning what constitutes a cellular origin of replication, information has come mostly from viral systems where the origins are defined genetically (6,10,21). Among the papova-and adenoviruses, the involvement of transcription regulatory elements in replication control is very well documented (12,25,31). Conversely, template replication is typically required for late gene transcription (9,19). Thus, there is important communication between control of replication and transcription.The elements jointly controlling DNA and RNA synthesis are generally located near the replication origin in these viruses (5,12,22). These are binding sites for different factors in the different viruses, demonstrating that a variety of factors may play dual regulatory roles. These factors can be derived from the host or encoded by the virus. The host proteins are known to function as transcription factors for cellular genes and include, for example, NFI/CTF, used by BK virus and adenovirus, GC box binding factors such as Spl, used by simian virus 40 (SV40), and a variety of cellular proteins that bind viral enhancer regions (14,25,26,28). Some proteins that influence both processes appear to have separate domains for transcription and replication stimulation (18,32,33,37 to stimulate both replication and transcription was demonstrated. Stimulation of either process required the fused transcription activation domain. There was also a correlation between the strength of transcription activation and that of replication activation (2). This raised the possibility that nearly any transcription factor might have the capacity to stimulate replication when bound near an origin. A potential mechanism for this was that the factor activation domain might interact with a common mediating factor required for both transcription and replication.A different mechanism was suggested by experiments with the host NFI factor used in BK transcription (7). Those results sh...

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Section: Resultsmentioning

confidence: 99%

The replication activation potential of selected RNA polymerase II promoter elements at the simian virus 40 origin.

Hoang¹,

Wang²,

Gralla³

1992

Mol. Cell. Biol.

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“…Cells (3610 6 ) were transfected with 30 mg of mutagen-treated pYZ289 and 40 mg of pECM53k or pECMmutk and incubated in 150-mm culture dishes for 72 h. Plasmid DNA was then isolated (Hirt, 1967), puri®ed by digestion with proteinase K (2 mg/ml) and phenol/chloroform extraction, treated with ribonuclease A (0.2 mg/ml) and digested with DpnI (Peden et al, 1980). Plasmids harvested from individual transfection dishes were puri®ed and analysed separately.…”

Section: Characterization Of Supf Mutantsmentioning

confidence: 99%

The p53 tumor suppressor protein reduces point mutation frequency of a shuttle vector modified by the chemical mutagens (±)7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, aflatoxin B1 and meta-chloroperoxybenzoic acid

Courtemanche

Anderson

1999

Oncogene

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The p53 tumor suppressor protein reduces point mutation frequency of a shuttle vector modi®ed by the chemical mutagens (+)7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, a¯atoxin B 1 and meta-chloroperoxybenzoic acid Chantal Courtemanche 1,2 and Alan Anderson* ,1,2 1 Centre de recherche en canceÂrologie de l'UniversiteÂ Laval, Pavillon L'HoÃtel-Dieu de QueÂbec, Centre hospitalier universitaire de QueÂbec, QueÂbec G1R 2J6 Canada; and 2 DeÂpartement de biologie, UniversiteÂ Laval, QueÂbec G1K 7P4 Canada p53 has been postulated to be the guardian of the genome. However, results supporting the prediction that point mutation frequencies are elevated in p53-de®cient cells either have not been forthcoming or have been equivocal. To analyse the e ect of p53 on point mutation frequency, we used the supF gene of the pYZ289 shuttle vector as a mutagenic target. pYZ289 was treated in vitro by ultraviolet irradiation, a¯atoxin B 1 , (+)7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene and meta-chloroperoxybenzoic acid and then transfected into p53-de®cient cells with or without a p53 expression vector. p53 reduced the mutant frequency up to ®vefold when pYZ289 was treated with a¯atoxin B 1 , (+)7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene or meta-chloroperoxybenzoic acid but not when it was ultraviolet-irradiated. The p53-dependent mutation frequency reduction was higher at a higher level of premutational lesions for a¯atoxin B 1 and (+)7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene and at a lower level of lesions for meta-chloroperoxybenzoic acid. This suggests that the chemical mutagens produce, in a dose-dependent fashion, two kinds of DNA damage, one subject to p53-dependent mutation frequency reduction and the other not. These results indicate that p53 can reduce the point mutation frequency in a shuttle vector treated by chemical mutagens and suggest that p53 can act as guardian of the genome for at least some kinds of point mutations.

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“…As judged from the hybridization intensity of standard plWl DNA, the number of molecules retained in the cells at 4 d after transfection was -100 per cell, and about eight molecules within a cell replicated during a cell generation. We examined replication of plWl in the absence of BrdU, on the basis of susceptibility to methylation-sensitive restriction enzymes, Dpn I and Mbo I (Peden et al, 1980;Li and Kelly, 1984). The Hirt extract was prepared from 293S cells at 4 d after cotransfection with plWl and pUC119 and treated with various concentrations of Dpn I.…”

Section: Replication Ofmentioning

confidence: 99%

Autonomous replication of human chromosomal DNA fragments in human cells.

Masukata

Satoh

Obuse

et al. 1993

MBoC

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We have examined whether a human chromosome has distinct segments that can replicate autonomously as extrachromosomal elements. Human 293S cells were transfected with a set of human chromosomal DNA fragments of 8-15 kilobase pairs that were cloned on an Escherichia coli plasmid vector. The transfected cells were subsequently cultured in the presence of 5-bromodeoxyuridine during two cell generations, and several plasmid clones labeled in both of the daughter DNA strands were isolated. Efficiency of replication of these clones, as determined from the ratios of heavy-heavy and one-half of heavy-light molecules to total molecules recovered from density-labeled cells, was 9.4% per cell generation on the average. Replication efficiency of control clones excluded during the selection was about 2.2% and that of the vector plasmid alone was 0.3%. A representative clone plWl replicated in a semiconservative manner only one round during the S phase of the cell cycle. It replicated extrachromosomally without integration into chromosome. The human segment of the clone was composed of several subsegments that promoted autonomous replication at different efficiencies. Our results suggest that certain specific nucleotide sequences are involved in autonomous replication of human segments.

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Isolation of Mutants of an Animal Virus in Bacteria

Cited by 343 publications

References 35 publications

The replication activation potential of selected RNA polymerase II promoter elements at the simian virus 40 origin.

The replication activation potential of selected RNA polymerase II promoter elements at the simian virus 40 origin.

The p53 tumor suppressor protein reduces point mutation frequency of a shuttle vector modified by the chemical mutagens (±)7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, aflatoxin B1 and meta-chloroperoxybenzoic acid

Autonomous replication of human chromosomal DNA fragments in human cells.

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