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Multiply branched replicative intermediates in E. coli and bacteriophage lambda.

M S Valenzuela, R B Inman

    Molecular & General Genetics : MGG
    |January 1, 1981
    PubMed
    Summary
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    Researchers isolated branched DNA fragments from E. coli, potentially forming during DNA replication under specific starvation conditions. These structures may arise from secondary initiation or rapid DNA strand re-annealing near replication forks.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Bacterial DNA replication involves complex structures and intermediates.
    • Synchronization of cell cycles in bacteria like E. coli often requires specific nutrient starvation.
    • Understanding DNA replication intermediates is crucial for comprehending genome stability.

    Purpose of the Study:

    • To isolate and characterize branched DNA fragments found in E. coli.
    • To investigate the origin and formation of these branched DNA structures.
    • To determine the association of these structures with cell synchronization procedures.

    Main Methods:

    • Isolation of fast sedimenting complexes containing branched DNA from E. coli.
    • Observation of similar structures in bacteriophage lambda DNA replication intermediates.

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  • Analysis of DNA structures following amino acid and thymidine starvation for synchronization.

    • Main Results:

    • Multiple branched DNA fragments were isolated in a fast sedimenting complex from E. coli.
    • These structures represent a small fraction of the E. coli genome.
    • Similar branched DNA structures were observed in bacteriophage lambda DNA replication intermediates in synchronized E. coli.

    Conclusions:

    • Branched DNA structures are associated with the synchronization process in E. coli.
    • These structures may originate from secondary replication initiation sites.
    • Alternatively, they could form via rapid snap-back of single-stranded daughter DNA strands near replication forks.