Analysis of the α‐Satellite DNA from African Green Monkey Cells by Restriction Nucleases

Fittler, Friedrich

doi:10.1111/j.1432-1033.1977.tb11399.x

Cited by 40 publications

(17 citation statements)

References 19 publications

(13 reference statements)

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“…The existence of different sorts of heterogeneities in satellite DNAs has now been widely confirmed (9,11,12). When present in almost all of the repetition units, the sites have been called type A while those present in only a small portion of them have been called type B (9).…”

Section: Introductionmentioning

confidence: 97%

The organisation of the long range periodicity calf satellite DNA I variants as revealed by restriction enzyme analysis

Roizes¹,

Pagès²,

Lecou³

1980

Nucl Acids Res

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The analysis of a large number of restriction sites within the long range periodicity calf satellite DNA I does not reveal a superimposable shorter repeat. Although some restriction sites are present in almost all the 100,000 tandemly arranged copies of the 1460 bp repetition unit, other sites such as Atu CI occur at much lower frequencies. When present they are distributed randomly along the satellite DNA molecules. The missing sites appear to result from random and presumably single base alterations. Digestion with the enzymes Hha I and Kpn I showed another type of variant to exist within the calf satellite DNA I. Unlike Atu CI the distributions of the variants detected by these enzymes are not random and organised on long stretches of satellite DNA. The possible functional significance and evolutionary implication of these results are discussed. INTRODUCTIONThe use of restriction endonucleases in the study of satellite DNAs has shown these highly repetitive sequences to posses a greater complexity than originally expected.

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Section: Introductionmentioning

confidence: 97%

The organisation of the long range periodicity calf satellite DNA I variants as revealed by restriction enzyme analysis

Roizes¹,

Pagès²,

Lecou³

1980

Nucl Acids Res

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“…Moreover, it has been reported that alpha satellite DNA is more abundant in some Cercopithecini species (up to 20% of the genome of Chlorocebus aethiops ) [36] than in great apes, where its contribution would reach only 3% of the genome [14]. Finally, enzymatic digestion of genomic DNA from various Old World monkey species can lead to a clear alpha satellite ladder pattern which is not observed when human or chimpanzee DNA is used, thereby pointing to different composition and organization of alpha satellite DNA in Old World monkeys [37]. …”

Section: Introductionmentioning

confidence: 99%

Diversity and distribution of alpha satellite DNA in the genome of an Old World monkey: Cercopithecus solatus

et al. 2016

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BackgroundAlpha satellite is the major repeated DNA element of primate centromeres. Evolution of these tandemly repeated sequences has led to the existence of numerous families of monomers exhibiting specific organizational patterns. The limited amount of information available in non-human primates is a restriction to the understanding of the evolutionary dynamics of alpha satellite DNA.ResultsWe carried out the targeted high-throughput sequencing of alpha satellite monomers and dimers from the Cercopithecus solatus genome, an Old World monkey from the Cercopithecini tribe. Computational approaches were used to infer the existence of sequence families and to study how these families are organized with respect to each other. While previous studies had suggested that alpha satellites in Old World monkeys were poorly diversified, our analysis provides evidence for the existence of at least four distinct families of sequences within the studied species and of higher order organizational patterns. Fluorescence in situ hybridization using oligonucleotide probes that are able to target each family in a specific way showed that the different families had distinct distributions on chromosomes and were not homogeneously distributed between chromosomes.ConclusionsOur new approach provides an unprecedented and comprehensive view of the diversity and organization of alpha satellites in a species outside the hominoid group. We consider these data with respect to previously known alpha satellite families and to potential mechanisms for satellite DNA evolution. Applying this approach to other species will open new perspectives regarding the integration of satellite DNA into comparative genomic and cytogenetic studies.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3246-5) contains supplementary material, which is available to authorized users.

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“…Two of the plasmids, 12-9 and (Fig. 4D), hybridized to the EcoRI-digested COS7 DNA in a ladder pattern characteristic of the a-satellite, a repeated sequence found in approximately 10,000 copies per haploid cell, both dispersed and in tandem arrays (5,32). That these inserts indeed contained a sequences was confirmed by their hybridization to an aspecific probe (data not shown).…”

Section: Methodsmentioning

confidence: 99%

Transfected DNA is mutated in monkey, mouse, and human cells.

Lebkowski¹,

DuBridge²,

Antell³

et al. 1984

Mol. Cell. Biol.

165

110

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Papovavirus-based shuttle vectors containing the bacterial lacI gene were used to show that a mutation frequency in the range of 1% occurs in lacI when such vectors are transfected into COS7 and CV-1 simian cells, NIH 3T3, 3T6, L, and C127 mouse cells, and human 293 and HeLa cells. This frequency is approximately four orders of magnitude higher than the spontaneous mutation frequency in either mammalian or bacterial cells. The mutations are predominantly base substitutions and deletions and also include insertions from the mammalian genome. Time course experiments argue that mutagenesis occurs soon after arrival of the DNA into the nucleus. However, replication of the vector is not required since mutations occur even when the vector lacks all viral sequences. The high mutation frequency appears to be the characteristic outcome of transfection of DNA into mammalian cells.Mammalian cells in culture readily take up DNA. Only a small portion of this transfected DNA reaches the nucleus, and yet a smaller fraction is stably incorporated into the chromosomes in a situation permitting its expression. Although the transfection process is poorly understood, transfection of DNA has become a tool of pivotal importance in the study of the molecular biology of mammalian cells. Application of the technique has facilitated study of topics such as the expression of manipulated genes and the identification of sequences with oncogenic potential. We have sought to use DNA transfection to study mutation in mammalian cells by incorporating an easily scored marker on a transfected vector. Previously, the lacd gene of Escherichia coli was used as the mutational target on simian virus 40 (SV40)-based vectors that could replicate in both COS7 simian cells and bacteria. We found that, after transfection by the DEAE-dextran technique and 2-day passage in COS7 cells, 1% of the plasmids rescued to E. coli contained mutations in lacI (3). A similar mutation frequency was detected in related vectors, using the galK marker (29). This strikingly high mutation frequency and the facility of the lacI system for detailed analysis of the mutations have led us to extend this approach by using different vectors and cell lines, to characterize further the mutational fate of transfected DNA.In this study, a series of vectors was introduced into a variety of mammalian cell lines to establish the generality of the high mutation frequency. A description of the host DNA that has in some cases become joined to the vectors is also included. The kinetics of mutagenesis were examined by performing time course experiments. To determine whether replication is required for mutagenesis, we introduced molecules lacking a viral origin of replication.Our observations argue that all transfected DNA, with or without any viral DNA, appears extremely susceptible to mutagenesis in mammalian cells. These findings are compatible with studies that indicate that cotransfected DNA species can become covalently joined, both by homologous recombination and by blunt-end ligation, a...

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Analysis of the α‐Satellite DNA from African Green Monkey Cells by Restriction Nucleases

Cited by 40 publications

References 19 publications

The organisation of the long range periodicity calf satellite DNA I variants as revealed by restriction enzyme analysis

The organisation of the long range periodicity calf satellite DNA I variants as revealed by restriction enzyme analysis

Diversity and distribution of alpha satellite DNA in the genome of an Old World monkey: Cercopithecus solatus

Transfected DNA is mutated in monkey, mouse, and human cells.

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