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To synthesize DNA, the double-stranded DNA is unwound by DNA helicases ahead of polymerases, forming a replication fork containing two single-stranded templates. Replication processes permit copying a single DNA double helix into two DNA helices, which are divided into the daughter cells at mitosis .
Eukaryotes initiate DNA replication at multiple points in the chromosome, so replication forks meet and terminate at many points in the chromosome. Because eukaryotes have linear chromosomes, DNA replication is unable to reach the very end of the chromosomes. Due to this problem, DNA is lost in each replication cycle from the end of the chromosome.
Before DNA replication can start, the pre-replicative complex assembles at origins to load helicase onto DNA. The complex assembles in late mitosis and early G1. Assembly of these pre-replicative complexes (pre-RCs) is regulated in a manner that coordinates DNA replication with the cell cycle. [6]
A helicase is an enzyme that plays a crucial role in the DNA replication and repair processes. Its primary function is to unwind the double-stranded DNA molecule by breaking the hydrogen bonds between the complementary base pairs, allowing the DNA strands to separate.
When Mcm2-7 is first loaded it completely encircles the DNA and helicase activity is inhibited. In S phase, the Mcm2-7 complex interacts with helicase cofactors Cdc45 and GINS to isolate a single DNA strand, unwind the origin, and begin replication down the chromosome. In order to have bidirectional replication, this process happens twice at an ...
A new MCM complex cannot be loaded onto the origin until the pre-RC subunits are reactivated with the decline of CDK activity at the end of mitosis. Thus, CDKs serve a dual role in the regulation of eukaryotic DNA replication: elevated CDK activity initiates replication at the origins and prevents rereplication by inhibiting origin re-licensing.
MCM2-7 is required for both DNA replication initiation and elongation; its regulation at each stage is a central feature of eukaryotic DNA replication. [3] During G1 phase, the two head-to-head Mcm2-7 rings serve as the scaffold for the assembly of the bidirectional replication initiation complexes at the replication origin.
A distinct group of DNA-binding proteins is the DNA-binding proteins that specifically bind single-stranded DNA. In humans, replication protein A is the best-understood member of this family and is used in processes where the double helix is separated, including DNA replication, recombination, and DNA repair. [123]