Lecture 13: Cell Cycle Control

Cell Cycle Control

Learning Objectives

  • Features checked at cell cycle checkpoints: Students will identify the specific features that are monitored at each checkpoint during the cell cycle.

  • Downstream effects of cell cycle regulator molecules: Students will learn to predict the consequences of both normal and abnormal functioning of these regulatory molecules.

  • Influence of cues on cell cycle regulators: Students will identify how various cues impact the activity of cell cycle regulator molecules.

  • Failure of cell cycle regulators leading to cancer: Students will be able to describe how the malfunction of cell cycle regulators contributes to the development of cancer.

Overview of Cancer

  • The discussion explicitly centers around the topic of cancer, which is a broad category of diseases characterized by uncontrolled cell growth.

Cell Division Rates and Types of Cells

  1. Intestinal Cells:

    • Go through two cell cycles per day.

    • Consideration: Which phases of the cell cycle are significantly shortened to achieve this rapid division?

  2. Nerve Cells:

    • (Almost) never divide.

    • Consideration: Which phase of the cell cycle do these cells predominantly remain in?

Cell Cycle Checkpoints

  • Checkpoints: Critical control mechanisms that ensure cells do not proceed through the cell cycle until specific conditions are met.

    • G1 Checkpoint:

    • Checks for:

      • Cell size

      • Nutrients

      • Growth factors

      • DNA damage

    • S (Synthesis) Phase: DNA replication occurs.

    • G2 Checkpoint:

    • Checks for:

      • DNA damage

      • Completeness of DNA replication

    • M (Mitosis) Checkpoint (Spindle Checkpoint):

    • Checks for:

      • Proper attachment of chromosomes to the spindle at the metaphase plate.

Conditions Checked by Specific Checkpoints

  • Microtubule attachment failure to kinetochores: Checked by M checkpoint.

  • Mutation due to DNA synthesis error: Checked by G2 checkpoint.

  • Mutation due to environmental stressors: Also checked by G2 checkpoint.

Checkpoint Regulators

  • Role of Regulator Molecules: Active during checkpoints; they prevent the division of damaged or otherwise compromised cells.

  • Outcomes if a cell is prevented from passing through a checkpoint:

    • The cell may:

    • Enter a resting state (G0)

    • Repair DNA damage

    • Undergo apoptosis (programmed cell death).

  • Consequences of defective regulator molecules: If checkpoint regulators are defective, cells may continue to divide uncontrollably, potentially forming tumors.

Regulator Molecules: Cyclins

  • Function of Cyclins: Cyclins are essential for prompting cell movement from one phase to the next. Key cyclins include:

    • G1 Cyclin (Cyclin D)

    • G1/S Cyclin (Cyclin E)

    • S Cyclin (Cyclin A)

    • M Cyclin (Cyclin B)

Regulator Molecules: Cyclins and Cdks

  • Cyclins operate by binding to Cyclin-dependent kinases (Cdks).

  • Function of Cdks: When activated, Cdks phosphorylate target proteins, altering their activity levels.

  • Mechanism of action:

    • When a cyclin binds to a Cdk, it activates the Cdk and directs it to specific proteins for phosphorylation.

Example of Target Proteins for Cyclin E-Cdk Complex

  • Cyclin E bound to Cdk: Phosphorylates and activates proteins that are crucial during the S phase of the cell cycle.

Cyclins Activate Cdks

  • Presence of Cdks: They are perpetually present but remain inactive without association with cyclins.

  • Phosphorylation of enzymes (e.g., helicase and topoisomerase): When active, these enzymes initiate S phase (DNA replication).

Regulator Molecules: M Phase Promoting Factor (MPF)

  • Composition: MPF is a complex of Cdk and Cyclin B.

  • Threshold Requirement: When Cyclin B concentrations are sufficiently high, MPF is formed.

  • Role of MPF: Catalyzes phosphorylation of proteins necessary for initiating M phase, such as:

    • Nuclear lamins for breaking down the nuclear envelope.

    • Microtubules for the assembly of the spindle apparatus.

Regulator Molecules: MAD (Mitotic Arrest Deficit Proteins)

  • Function at M/Spindle Checkpoint:

    • Checks for chromosomal attachment to the spindle.

    • Inhibition of Anaphase-Promoting Complex/Cyclosome (APC/C) while waiting for correct chromosomal attachments.

  • Consequences of prolonged incorrect attachment: This condition is termed Mitotic Catastrophe; it may initiate apoptotic pathways via Bcl-2 family proteins.

  • If attachments are correct: MAD's inhibition of APC/C ceases, allowing MPF to activate APC/C, leading to anaphase.

Regulator Molecules: Anaphase-Promoting Complex/Cyclosome (APC/C)

  • Function: When chromosomes are correctly attached:

    • APC/C inhibition by MAD is lifted, leading to the destruction of regulatory proteins via ubiquitination.

  • Ubiquitin tagging: This process signals degradation of target proteins, which ceases their functions.

APC/C, Separase, and Securin Interaction

  1. APC/C and Cyclin B: At the conclusion of mitosis, APC/C ubiquitinates Cyclin B, resulting in:

    • (a) Destruction of MPF, (b) Activation of MPF.

  2. Separase role: Separase, when bound to Securin, is inactive; APC/C ubiquitinates Securin, leading to:

    • (a) Destruction of Securin, (c) Activation of Separase.

Role of Activated Separase

  • Separase cleaves the cohesin complex that joins sister chromatids, facilitating their separation and the completion of mitosis.

Cancer and Cell Division

  • Characteristics of Cancer Cells: Often exhibit abnormal chromosome numbers and structures, leading to uncontrolled proliferation.

  • Mutations Leading to Tumor Development:

    • Examples: Mutations in negative regulators (e.g., P53 and MAD) may result in their inactivity, while positive regulators (e.g., cyclins) may become overactive.

Regulation of Cdks, Cyclins, and APC/C by Cues

  • Activity of these regulators is responsive to internal and external cues. Examples include:

    1. Hormonal Growth Factors: In normally functioning cells, these factors lead to:

    • (a) Increased activity of Cdks and cyclins.

    1. DNA Damage: Typically results in:

    • (b) Decreased activity of Cdks and cyclins.

P53 Tumor Suppressor Gene

  • Function of p53: Encodes the p53 protein, which detects DNA damage at the G1 checkpoint.

  • Mechanism of Action:

    • When active, p53 acts as a transcription factor that induces the synthesis of Cyclin-dependent kinase inhibitor proteins (CKIs).

    • CKIs inhibit the cyclin/Cdk complexes, effectively pausing the cell cycle and allowing time for DNA repair.

    • Activates DNA repair mechanisms, such as nucleotide excision repair, during the G2 checkpoint.

P53 and Cellular Outcomes

  • If Damage Cannot Be Repaired: P53 triggers:

    • Apoptosis, leading to:

    • DNA fragmentation.

    • Organelle fragmentation.

    • Fragmentation of the cell into smaller pieces that signal for immune clearance.

Comparison of Normal vs Nonfunctional P53

  • Normal p53: Activates following DNA damage, leading to CKI production that halts cell cycle progression.

  • Nonfunctional p53: Cannot bind to DNA, leading to continuing cell cycle progression despite DNA damage, risking the transmission of damaged DNA.

Cancer: Out-of-Control Cell Division

  • Commonality Across Cancers: All cancers arise from cells with multiple failures in cell cycle regulation.

  • Predicted Characteristics of Cancer Cells:

    1. Negative regulators like the P53 and MAD will typically show:

    • (a) Inactivity.

    1. Positive regulators such as cyclins will likely present:

    • (b) Overactivity.

Genes and Oncogenes

  • Mutations Leading to Oncogenes: Recapitulates how normal genes that encode proteins involved in cell division can mutate into oncogenes.

  • Ras Oncogene Example:

    • Healthy cell dynamics state: In absence of growth factors, Ras remains:

    • (a) Inactive, (c) Does not divide.

    • Cancer cell dynamics: Following mutation, Ras remains:

    • (b) Overactive, (d) Divides uncontrollably.

Consequences of Cancer Cell Division

  • Impact of Cancer Cells: Cells divide in an uncontrolled manner, infiltrating and impairing healthy tissues, consuming vital nutrients and disrupt the function of normal cellular structures.

  • Diversity of Cancers: There are at least 200 recognized types of cancer, each with distinct characteristics and implications for affected individuals.