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PubMed · 6089891

Replication pattern of human repeated DNA sequences.

R Meneveri·A Agresti·D Breviario·E Ginelli
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Abstract

Either aphidicolin- or thymidine-synchronized human HL-60 cells were used to study the replication pattern of a family of human repetitive DNA sequences, the Eco RI 340 bp family (alpha RI-DNA), and of the ladders of fragments generated in total human DNA after digestion with XbaI and HaeIII (alpha satellite sequences). DNAs replicated in early, middle-early, middle-late and late S periods were labelled with BUdR or with [3H]thymidine. The efficiency of the cell synchronization procedure was confirmed by the transition from a high-GC to a high-AT average base composition of the DNA synthesized going from early to late S periods. By hybridizing EcoRI 340 bp repetitive fragments to BUdR-DNAs it was found that this family of sequences is replicated throughout the entire S period. Comparing fluorograph densitometric scans of [3H]DNAs to the scans of ethidium bromide patterns of total HL-60 DNA digested with XbaI and HaeIII, it was observed that DNA synthesized in different S periods is characterized by approximately the same ladder of fragments, while the intensity of each band may vary through the S phase; in particular, the XbaI 2.4 kb fragment becomes undetectable in late S.

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Base SequenceCell LineCentrifugation, Density GradientDNADNA Restriction EnzymesDeoxyribonuclease EcoRIDeoxyribonucleases, Type II Site-SpecificHumansLeukemia, MyeloidNucleic Acid ConformationNucleic Acid HybridizationRepetitive Sequences, Nucleic AcidJournal ArticleResearch Support, Non-U.S. Gov't

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R Meneveri, A Agresti, D Breviario, E Ginelli. 1984-10-05. Replication pattern of human repeated DNA sequences.. https://doi.org/10.1016/0167-4781(84)90077-0

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