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    Area of Science:

    • Molecular Biology
    • Genetics
    • Biophysics

    Background:

    • Genomes organize hierarchically, with intracellular phase transitions compartmentalizing processes like transcription and DNA repair.
    • The role of phase transitions in DNA replication, particularly coordinating double- and single-stranded DNA, remains largely unexplored.

    Purpose of the Study:

    • To investigate the molecular interactions driving the condensation of mitochondrial replication components into nucleoids.
    • To understand how phase transitions contribute to the organization and regulation of mitochondrial DNA replication.

    Main Methods:

    • Purified mitochondrial replication components were used to observe phase behavior.
    • Single-molecule experiments were conducted to analyze protein-DNA interactions within condensates.
    • A minimalistic coarse-grained model was developed to simulate component assembly.

    Main Results:

    • Purified mitochondrial replication components exhibited complex multiphasic behavior, with nucleic acids colocalizing with cognate proteins.
    • Single-stranded DNA formation increased the partitioning of its cognate protein (mtSSB) within condensates.
    • The model demonstrated how pairwise interactions dictate assembly within condensates, explaining preferential mtSSB localization to replicating sites.

    Conclusions:

    • Mitochondrial nucleoids form through multiphasic condensates driven by specific molecular interactions.
    • The findings suggest that phase transitions play a crucial role in organizing mitochondrial DNA replication.
    • This provides mechanistic insights into the spontaneous organization of replication within cells.