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Related Concept Videos

Restarting Stalled Replication Forks02:37

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DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Conservative Site-specific Recombination and Phase Variation02:53

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I,...
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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
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Nucleosomes are the basic units of chromatin compaction. Each nucleosome consists of the DNA bound tightly around a histone core, which makes the DNA inaccessible to DNA binding proteins such as DNA polymerase and RNA polymerase. Hence, the fundamental problem is to ensure access to DNA when appropriate, despite the compact and protective chromatin structure.
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Related Experiment Video

Updated: Sep 11, 2025

Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark
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RIF1 orchestrates a multi-step restoration of post-replicative H3K9me3.

Naiming Chen, Susanne Bandau, Reshma Ravindran Nair Pushkala Kumari

    Biorxiv : the Preprint Server for Biology
    |August 12, 2025
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    Summary
    This summary is machine-generated.

    RIF1 protein helps re-establish histone marks after DNA replication. It recruits key enzymes to restore H3K9me3 levels during the G1 phase, ensuring proper epigenetic identity.

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    Chromatin Immunoprecipitation ChIP of Histone Modifications from Saccharomyces cerevisiae
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    Area of Science:

    • Epigenetics
    • Molecular Biology
    • Cell Cycle Regulation

    Background:

    • DNA replication dilutes parental histones and their modifications.
    • Restoration kinetics of histone marks determine epigenetic identity regain post-replication.
    • H3K9me3 restoration is slow, continuing through G1 phase, with unknown mechanisms.

    Purpose of the Study:

    • To elucidate the molecular mechanisms behind the slow H3K9me3 restoration after DNA replication.
    • To investigate the role of RIF1 in the H3K9me3 reconstitution process.

    Main Methods:

    • Chromatin immunoprecipitation assays.
    • Analysis of histone modifications (H3K9me3, H3S10ph).
    • Investigating protein-protein interactions (RIF1, SUV39H1, HP1, PP1α, Aurora kinase).

    Main Results:

    • RIF1 reassociation with heterochromatin at mitotic exit is crucial for H3K9me3 deposition.
    • RIF1 recruits SUV39H1, HP1α, and HP1β, promoting H3K9 tri-methylation in G1.
    • RIF1 recruits PP1α, and their interaction is vital for maintaining H3K9me3 levels.
    • RIF1-PP1 complex restrains Aurora kinase, preventing premature H3S10 phosphorylation and allowing H3K9me3 reinstatement.

    Conclusions:

    • RIF1 orchestrates heterochromatin re-establishment post-replication.
    • RIF1 ensures timely H3K9me3 restoration by coordinating histone methyltransferases and phosphatases.
    • RIF1 acts as a critical regulator for epigenetic memory maintenance across cell divisions.