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Related Concept Videos

S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
Separation of Sister Chromatids02:17

Separation of Sister Chromatids

At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
Chromosome Structure02:40

Chromosome Structure

A functional eukaryotic chromosome must contain three elements: a centromere, telomeres, and numerous origins of replication.
The centromere is a DNA sequence that links sister chromatids. This is also where kinetochores, protein complexes to which spindle microtubules attach, are constructed after the chromosome is replicated. The kinetochores allow the spindle microtubules to move the chromosomes within the cell during cell division.
Telomeres consist of non-coding repetitive nucleotide...

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Determination of S-Phase Duration Using 5-Ethynyl-2'-deoxyuridine Incorporation in Saccharomyces cerevisiae
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Structural changes in Mcm5 protein bypass Cdc7-Dbf4 function and reduce replication origin efficiency in

Margaret L Hoang1, Ronald P Leon, Luis Pessoa-Brandao

  • 1Department of Genome Sciences, University of Washington, Seattle, USA.

Molecular and Cellular Biology
|August 29, 2007
PubMed
Summary

The mcm5-bob1 mutation in yeast allows DNA replication origins to fire without Dbf4-dependent kinase (DDK) but reduces efficiency. This suggests Mcm5 protein

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Area of Science:

  • Cellular biology
  • Molecular genetics
  • Biochemistry

Background:

  • Eukaryotic DNA replication initiates from multiple origins during S phase.
  • Prereplication complexes assemble on all origins in G1, but only a subset is activated by DDK (Dbf4-dependent kinase).

Purpose of the Study:

  • To investigate the role of Mcm5 protein in regulating origin firing efficiency and DDK-dependent activation.
  • To understand the mechanism by which the mcm5-bob1 mutation bypasses DDK control.

Main Methods:

  • Utilized the yeast mcm5-bob1 (P83L) mutation to study origin firing.
  • Analyzed origin efficiency at endogenous loci and on minichromosomes.
  • Investigated the structural and functional consequences of the mutation, referencing archaeal MCM homologue structures.

Main Results:

  • The mcm5-bob1 mutation bypasses DDK-mediated activation but reduces intrinsic origin firing efficiency.
  • This suggests Mcm5 protein adopts altered conformations, impacting origin activation.
  • A suppressor mutation interacting with P83L supports the conformational change hypothesis.

Conclusions:

  • DDK phosphorylation likely enforces a single, active Mcm5 conformation for efficient origin firing.
  • The mcm5-bob1 mutation allows multiple Mcm5 conformations, only one of which is permissive for activation, explaining DDK bypass and reduced efficiency.
  • Mcm5 appears to be a unique and direct target of DDK regulation in the minichromosome maintenance complex.