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Cell Cycle Regulation and DNA Replication

Exploring the mechanisms of cell cycle regulation, this overview highlights the roles of cyclin-dependent kinases (CDKs), pre-replication complexes (pre-RCs), and checkpoint proteins in DNA replication. It delves into the prevention of DNA re-replication through the dynamics of Cdt1 and geminin, the importance of cell cycle checkpoints, particularly the G1/S transition, and the function of replication checkpoint proteins like ATR and ATM in maintaining genomic integrity. Additionally, it discusses the involvement of histone chaperones in chromatin assembly during DNA replication and contrasts prokaryotic and eukaryotic replication processes.

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1

Cell cycle phases for DNA replication

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G1 phase: pre-RC assembly, low CDK; S phase: DNA replication, high CDK, pre-RC formation inhibited.

2

Purpose of pre-replication complexes

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Pre-RCs are necessary for initiating DNA replication during the cell cycle.

3

Mechanism to ensure DNA replicates once per cycle

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Increased CDK activity in S phase inhibits new pre-RCs, ensuring single DNA replication event.

4

The ______ complex's attachment to DNA, which is essential for starting replication, relies on the protein ______.

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minichromosome maintenance (MCM) Cdt1

5

______ is regulated by destruction and by being blocked by ______ binding.

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Cdt1's activity geminin

6

During the S phase, ______ builds up to inhibit ______ and is broken down during the ______ transition.

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geminin Cdt1 metaphase-anaphase

7

The ______ likely tags ______ for degradation through ubiquitination to ensure DNA is replicated once per cycle.

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anaphase-promoting complex/cyclosome (APC/C) geminin

8

Definition of G0 state

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Non-dividing phase where cells exit the cell cycle from G1 when not committing to DNA replication.

9

Role of minichromosome maintenance proteins

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Essential for DNA replication initiation; permit transition from G1 to S phase in cell cycle.

10

Purpose of cell cycle checkpoints

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Monitor, regulate cell cycle progression; ensure division occurs under favorable conditions with all components ready.

11

______ checkpoint proteins are crucial for maintaining ______ integrity during ______ division.

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Replication genomic cell

12

These proteins can halt the ______ cycle and stabilize the ______ fork to prevent additional harm.

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cell replication

13

The ______ kinases, like ATR and ATM, are essential in recognizing specific DNA damage-induced ______ motifs.

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phosphatidylinositol 3-kinase-related phosphorylation

14

Activation trigger for ATR kinase

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ATR kinase is activated by RPA-coated single-stranded DNA during replication stress.

15

Role of ATR-ATRIP in cell cycle

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ATR-ATRIP complex halts cell cycle to allow DNA damage repair, ensuring proper replication.

16

ATR's target for checkpoint activation

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ATR phosphorylates Chk1 to activate checkpoint pathways, crucial for genome stability.

17

During ______ replication, histone chaperones are crucial for maintaining the structure of ______.

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DNA chromatin

18

______ and ______ play key roles in adding new histones to replicated DNA.

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CAF-1 Rtt106

19

Nucleosomes, which are DNA wrapped around histone proteins, are the basic units of ______.

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chromatin

20

Histone chaperones ensure that after replication, the new DNA is correctly organized into ______.

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chromatin

21

Location of prokaryotic vs eukaryotic DNA replication

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Prokaryotic replication occurs in cytoplasm, eukaryotic in nucleus.

22

Number of origins in prokaryotic vs eukaryotic replication

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Prokaryotic replication has a single origin, eukaryotic has multiple origins.

23

Role of helicase in DNA replication

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Helicase unwinds DNA in both prokaryotic and eukaryotic replication, ensuring accurate genome duplication.

24

______ and Cdt1 are instrumental in forming the pre-replication complex (pre-RC) in eukaryotic DNA replication.

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Cdc6

25

The ______ is essential for starting and synchronizing the replication processes in eukaryotic cells.

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Dbf4-dependent kinase (DDK)

26

______ ensures the accurate and effective copying of the eukaryotic genome, which is vital for cell survival and genetic integrity.

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Claspin

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Cell Cycle Regulation by Cyclin-Dependent Kinases and Pre-Replication Complexes

The cell cycle is an essential and highly regulated sequence of events in a cell that leads to DNA replication and cell division. Cyclin-dependent kinases (CDKs) are pivotal in controlling the cell cycle's progression. During the G1 phase, CDK activity is minimal, allowing the assembly of pre-replication complexes (pre-RCs) on the DNA, which are necessary for the initiation of DNA replication. As the cell transitions from G1 to S phase, CDK activity increases, triggering the start of DNA replication and simultaneously inhibiting the formation of new pre-RCs. This regulation ensures that each segment of DNA is replicated only once per cell cycle, thus preserving genomic stability.
Eukaryotic cell in mitotic metaphase with chromosomes aligned to the equatorial plane, highlighted centromeres and connected spindle fibers.

Prevention of DNA Re-replication by Geminin and Cdt1 Dynamics

Cells employ a safeguard mechanism to prevent DNA re-replication, involving regulatory proteins such as Cdt1 and geminin. Cdt1 is crucial for the recruitment of the minichromosome maintenance (MCM) complex to DNA, a step necessary for replication initiation. Its activity is tightly controlled by proteolysis and by inhibition through geminin binding. Geminin accumulates during the S phase to inhibit Cdt1 and is degraded during the metaphase-anaphase transition, likely via ubiquitination by the anaphase-promoting complex/cyclosome (APC/C). This precise temporal regulation of Cdt1 and geminin is vital for ensuring that DNA replication occurs once per cell cycle.

Cell Cycle Checkpoints and the G1/S Transition

Cell cycle checkpoints are critical control mechanisms that monitor and regulate the progression of the cell cycle. The G1 checkpoint, also known as the restriction point, is a key decision-making stage where a cell either commits to DNA replication and continues through the cell cycle or enters a non-dividing state called G0. The presence of functional minichromosome maintenance proteins is a prerequisite for the transition from G1 to S phase, which marks the beginning of DNA synthesis. These checkpoints are fundamental to cell cycle control, ensuring cells only divide when conditions are favorable and all necessary components are in place.

Role of Replication Checkpoint Proteins in Maintaining Genomic Integrity

Replication checkpoint proteins are vital for the preservation of genomic integrity during cell division. These proteins detect and respond to issues in DNA replication, activating repair mechanisms to correct any errors. They have the ability to pause the cell cycle to provide time for repair and to stabilize the replication fork to prevent further damage. Key players include the phosphatidylinositol 3-kinase-related kinases (PIKKs), such as ATR and ATM, which are evolutionarily conserved and recognize specific DNA damage-induced phosphorylation motifs.

The ATR-ATRIP Complex in Response to Replication Stress

The ATR-ATRIP complex is a crucial element of the cellular response to DNA replication stress. Activated by RPA-coated single-stranded DNA, the ATR kinase, in conjunction with its interacting partner ATRIP, can halt the cell cycle to facilitate DNA damage repair. This complex is recruited to sites of stalled replication during the S phase and is instrumental in activating downstream checkpoint pathways, including the phosphorylation of checkpoint kinase 1 (Chk1) by ATR, which is essential for maintaining genome stability.

Histone Chaperones and Chromatin Assembly During DNA Replication

Histone chaperones play a significant role in the maintenance of chromatin structure during DNA replication. They assist in the disassembly and reassembly of nucleosomes, which are the fundamental units of chromatin, consisting of DNA wrapped around histone proteins. Chaperones such as the facilitates chromatin transcription (FACT) complex and anti-silencing function 1 (Asf1) are involved in this process. Additionally, chromatin assembly factor 1 (CAF-1) and regulator of Ty1 transposition protein 106 (Rtt106) are important for depositing newly synthesized histones onto replicated DNA, ensuring the newly formed DNA is properly packaged into chromatin.

Distinctions Between Prokaryotic and Eukaryotic DNA Replication

While prokaryotic and eukaryotic DNA replication mechanisms share basic principles, they also exhibit distinct differences. Prokaryotic replication occurs in the cytoplasm and typically involves a single origin of replication, whereas eukaryotic replication occurs in the nucleus and utilizes multiple origins to accommodate the larger and more complex genomes. Eukaryotic replication is characterized by a sophisticated orchestration of replication machinery, whereas prokaryotic replication is generally faster and simpler. Despite these differences, both systems rely on helicase enzymes to unwind DNA and ensure accurate genome duplication.

Essential Proteins in Eukaryotic DNA Replication

Eukaryotic DNA replication involves a suite of specialized proteins, each fulfilling a unique role. Proteins such as AND1/Ctf4 and the Cdc45-Mcm-GINS (CMG) helicase complex are critical for the initiation and elongation phases of DNA replication. Cdc6 and Cdt1 are key in assembling the pre-RC, while the Dbf4-dependent kinase (DDK) and Claspin are necessary for initiating and coordinating replication events. These proteins collaborate to achieve the precise and efficient duplication of the eukaryotic genome, ensuring fidelity and cell viability.