Actin Polymerization and Muscle Contraction

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Questions and Answers

What is the significance of the sigmoidal curve in actin polymerization?

  • It indicates that actin polymerization is a slow and steady process.
  • It implies that actin polymerization requires a constant supply of energy.
  • It demonstrates that actin polymerization is directly proportional to the concentration of actin monomers.
  • It suggests that actin polymerization is a complex process with multiple steps. (correct)

Why does the addition of nucleated actin accelerate actin polymerization?

  • Nucleated actin increases the concentration of actin monomers, thus accelerating polymerization.
  • Nucleated actin provides a template for further polymerization, increasing the rate of monomer addition. (correct)
  • Nucleated actin acts as a catalyst, speeding up the reaction without being consumed.
  • Nucleated actin provides a stable environment for actin polymerization, minimizing the rate of depolymerization.

What is the primary reason why actin dimers and trimers are considered unstable?

  • They lack the sufficient number of hydrophobic interactions required for stability. (correct)
  • They are prone to rapid depolymerization due to their small size.
  • They are easily broken down by cellular enzymes, preventing their accumulation.
  • They lack the ability to bind ATP, which is essential for maintaining their structural integrity.

How does the use of Pyrene-conjugated actin contribute to the understanding of actin polymerization?

<p>Pyrene fluorescence provides a quantitative measure of the amount of actin polymer formed. (D)</p> Signup and view all the answers

What is the role of salt in actin polymerization?

<p>Salt promotes the interaction between actin monomers, leading to filament formation. (A)</p> Signup and view all the answers

Based on the information provided, what can be inferred about the role of ATP in actin polymerization?

<p>ATP is hydrolyzed during actin polymerization, providing energy for filament growth. (C)</p> Signup and view all the answers

Which of the following statements accurately describes the relationship between actin, myosin, and ATP in muscle contraction?

<p>ATP hydrolysis by myosin provides the energy for the movement of actin filaments along myosin. (C)</p> Signup and view all the answers

Based on the information provided, how does the role of actin in muscle contraction align with its role in cell motility?

<p>Actin plays a similar role in both, providing a dynamic framework for movement. (A)</p> Signup and view all the answers

What does the presence of an ATP/GTP cap at the plus end of an actin or microtubule filament indicate?

<p>The filament is undergoing rapid polymerization. (A)</p> Signup and view all the answers

What happens when a subunit in an actin or microtubule filament hydrolyzes ATP or GTP?

<p>The subunit becomes more susceptible to depolymerization. (C)</p> Signup and view all the answers

In vitro, what is the primary factor influencing the rate of actin filament polymerization?

<p>The concentration of ATP-bound G-actin monomers. (C)</p> Signup and view all the answers

Which of the following best describes the relationship between ATP/GTP hydrolysis and the stability of actin or microtubule filaments?

<p>Hydrolysis destabilizes filaments by promoting subunit dissociation. (B)</p> Signup and view all the answers

Why is the minus end of an actin or microtubule filament considered the slower growing end?

<p>The minus end is associated with a lower concentration of ATP/GTP-bound subunits. (B)</p> Signup and view all the answers

What is the primary role of ATPase or GTPase activity in the context of actin and microtubule dynamics?

<p>To regulate the rate of filament depolymerization by weakening inter-subunit bonds. (A)</p> Signup and view all the answers

What is the significance of the discovery of actin by Albert Szent-Györgyi's lab?

<p>It opened up new avenues for understanding the molecular basis of muscle contraction. (B)</p> Signup and view all the answers

How do actin and microtubule filaments differ in their dynamic behaviors?

<p>Actin filaments undergo more rapid depolymerization than microtubule filaments. (A)</p> Signup and view all the answers

At steady-state, what is the relationship between the free subunit concentration and the rate of polymer elongation?

<p>The elongation rate is zero. (D)</p> Signup and view all the answers

What does the critical concentration (Cc) represent in terms of polymer dynamics?

<p>The concentration of free subunits at which the polymer is in equilibrium. (A)</p> Signup and view all the answers

If the free subunit concentration is greater than the critical concentration (Cc), what happens to the polymer?

<p>The polymer grows. (B)</p> Signup and view all the answers

What is the mathematical expression for the elongation rate in terms of the free subunit concentration (subunit), the rate constant for association (kon), and the rate constant for dissociation (koff)?

<p>Elongation rate = kon * subunit - koff (C)</p> Signup and view all the answers

How does the polarity of actin affect its dynamics?

<p>The polarity of actin allows for bidirectional growth. (A)</p> Signup and view all the answers

What does the term "steady-state" refer to, in the context of cytoskeletal dynamics?

<p>A state where the rate of subunit addition is equal to the rate of subunit dissociation. (D)</p> Signup and view all the answers

What is the mathematical expression for the critical concentration ("Cc") in terms of the rate constants for association (kon) and dissociation (koff)?

<p>Cc = koff / kon (A)</p> Signup and view all the answers

What is the relationship between the free subunit concentration (subunit) and the rate of polymer elongation (dn/dt) when the subunit concentration is less than the critical concentration (Cc)?

<p>The elongation rate is negative and proportional to the difference between the subunit concentration and the critical concentration. (B)</p> Signup and view all the answers

If the free subunit concentration is equal to the critical concentration (Cc), what is the rate of polymer elongation (dn/dt)?

<p>dn/dt is zero. (B)</p> Signup and view all the answers

What does the term "free subunit" refer to in the context of cytoskeletal dynamics?

<p>A subunit that is not bound to the polymer. (D)</p> Signup and view all the answers

What is the critical concentration (Cc) in the context of actin filament dynamics?

<p>The concentration of actin monomers at which filament polymerization and depolymerization are balanced. (D)</p> Signup and view all the answers

Which of the following statements correctly describes the relationship between actin subunit concentration and filament dynamics?

<p>If the G-actin concentration is between the critical concentrations for the plus and minus ends, only the plus end will grow. (A)</p> Signup and view all the answers

What is the significance of nucleotide hydrolysis in actin filament dynamics?

<p>Nucleotide hydrolysis promotes dissociation of actin monomers from the filament. (D)</p> Signup and view all the answers

In the context of actin filament dynamics, which end of the filament is considered the 'growing' end?

<p>The plus end. (D)</p> Signup and view all the answers

What is the primary mechanism by which actin treadmilling occurs?

<p>Actin monomers are added to the plus end and removed from the minus end of the filament. (C)</p> Signup and view all the answers

How does actin treadmilling contribute to the motility of Listeria monocytogenes?

<p>Actin treadmilling provides a mechanism for the bacteria to propel itself through the cytoplasm of a host cell. (D)</p> Signup and view all the answers

Which of these is NOT a common property of actin and microtubule dynamics?

<p>Both have two distinct ends with different critical concentrations. (C)</p> Signup and view all the answers

Which of the following is NOT a state associated with actin filament dynamics?

<p>Degradation. (A)</p> Signup and view all the answers

Which of the following statements accurately describes the relationship between critical concentration (Cc) and elongation rate?

<p>Cc is inversely proportional to the elongation rate, meaning a higher Cc leads to a faster elongation rate. (C)</p> Signup and view all the answers

Based on the provided information, which statement accurately compares the elongation rate at the minus end versus the plus end of a filament?

<p>The relative elongation rates at the minus and plus ends can vary depending on the specific values of $k_{on}$ and $k_{off}$ at each end. (C)</p> Signup and view all the answers

If the critical concentration at the minus end ($Cc^{-}$) is greater than the critical concentration at the plus end ($Cc^{+}$), what is the expected behavior of the filament in this scenario?

<p>The filament will shrink at the minus end and grow at the plus end, effectively moving the filament in a specific direction. (D)</p> Signup and view all the answers

Why is the concept of treadmilling important in understanding filament dynamics?

<p>Treadmilling allows filaments to maintain a constant length despite continuous subunit turnover. (B)</p> Signup and view all the answers

Which of the following factors directly influences the values of $k_{on}$ and $k_{off}$ at each end of the filament?

<p>All of the above factors influence the values of $k_{on}$ and $k_{off}$. (D)</p> Signup and view all the answers

The difference in critical concentration between the plus and minus ends of a filament can be attributed to which of the following?

<p>All of the above factors contribute to the difference in critical concentration. (D)</p> Signup and view all the answers

Based on the provided information, what is the correct mathematical equation for calculating the critical concentration at the minus end ($Cc^{-}$)?

<p>$Cc^{-} = k_{off}/k_{on}$ (C)</p> Signup and view all the answers

Why is the statement "Elongation rate indicates net growth or shrink. This is not the same as the polymerization rate." important in understanding filament dynamics?

<p>It highlights that elongation rate, considering both polymerization and depolymerization, is a more accurate measure of filament dynamics than polymerization rate alone. (B)</p> Signup and view all the answers

Which component of the cytoskeleton is described as a hollow cylinder?

<p>Microtubule (C)</p> Signup and view all the answers

What is a common function of intermediate filaments?

<p>Mechanical support (A)</p> Signup and view all the answers

Which protein is not typically associated with intermediate filaments?

<p>α-tubulin (C)</p> Signup and view all the answers

Which cytoskeletal filament is primarily composed of G-actin subunits?

<p>Actin filament (D)</p> Signup and view all the answers

What structure is likely to have a functional equivalent of a nuclear lamina in plant cells?

<p>Cytoskeleton (D)</p> Signup and view all the answers

What is the primary role of microtubules in a cell?

<p>Transporting organelles (C)</p> Signup and view all the answers

Which cytoskeletal component is responsible for the structure and flexibility of epithelial cells?

<p>Intermediate filaments (D)</p> Signup and view all the answers

What type of cytoskeletal filament is least likely to be found in plant cells?

<p>Intermediate filament (C)</p> Signup and view all the answers

Flashcards

Microtubules

Hollow cylinders that are part of the cytoskeleton and help in cell shape and division.

Intermediate Filaments

Rope-like fibers that provide mechanical support to cells and stabilize their structures.

Cytoskeleton

A network of fibers composed of proteins that give shape and support to cells.

Actin Filament (F-actin)

A polymer made from globular actin (G-actin) that plays a role in cell movement and shape.

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Microtubule Composition

Composed of α-tubulin and β-tubulin, forming long hollow tubes in cells.

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Nuclear Lamina

A mesh-like structure made of intermediate filaments that provides support to the nucleus of a cell.

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Keratin

A type of intermediate filament found in epithelial cells that contributes to skin and hair structure.

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Dynamics of F-actin and Microtubules

Their properties include growth, shrinkage, and resilience in cellular activities.

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Nucleotide hydrolase

Enzymes that hydrolyze nucleotides, like ATP and GTP.

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G-actin

Globular form of actin that polymerizes to form F-actin.

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F-actin

Filamentous form of actin, made from G-actin subunits.

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ATPase

Enzyme that hydrolyzes ATP to ADP and Pi, involved in actin dynamics.

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GTPase

Enzyme that hydrolyzes GTP to GDP and Pi, crucial for microtubule dynamics.

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ATP/GTP cap

Stabilizing structure at the growing end of actin or microtubule, promoting growth.

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Polymerization

Process of monomers joining to form polymers, such as actin filaments.

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Depolymerization

Process where polymers break down into monomers, like actin filament disassembly.

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Actin polymerization

The process by which G-actin monomers form F-actin filaments.

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ATPase activity of Actin

The ability of actin to hydrolyze ATP, providing energy for muscle contraction.

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Nucleation

The initial step in actin polymerization that is rate-limiting.

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Fluorescent reporters in Actin studies

Tools, like pyrene conjugated G-actin, used to monitor actin polymerization.

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Sigmoidal reaction curve

The characteristic curve of actin polymerization, showing a lag phase before rapid growth.

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Stable dimers and trimers

Unstable initial structures that can lead to actin polymerization.

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Pyrene-actin signal

Indicates polymerized actin; its fluorescence is much higher in this form.

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Role of salt in polymerization

Salt influences the reversible polymerization of actin.

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Elongation Rate

The rate at which a polymer grows, defined by a simple equation involving rate constants for incorporation and dissociation.

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Rate Constant

A numerical value that quantifies the rate of incorporation or dissociation in polymer dynamics.

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Critical Concentration (Cc)

The concentration of free subunits at which there is no net growth or shrinkage of the polymer.

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Steady-State

A state during polymer dynamics where the rate of subunit addition equals the rate of subunit loss.

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Polymer Growth

Occurs when the concentration of subunits exceeds the critical concentration.

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Polymer Shrinkage

Occurs when subunit concentration is below critical concentration.

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Velocity in Calculus

The rate of change in distance over time, either constant or varying.

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Incorporation and Dissociation

Processes describing how subunits are added to or removed from a polymer.

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Net Change

The difference in the number of subunits over time, which determines polymer dynamics.

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Polarity of Actin

The characteristic of actin filaments that have a directionality in their growth.

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Filament Growth Rate

The speed at which filaments grow at their ends, influenced by various factors.

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Elongation Rate at Minus End

The rate at which filaments grow at the minus end, dependent on k-on and k-off rates.

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Elongation Rate at Plus End

The rate at which filaments grow at the plus end, higher than the minus end due to a faster k-on.

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Treadmilling

A dynamic state where subunits are added at one end and lost at the other, keeping filament length constant.

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ATP Cap

A structure formed by ATP-bound subunits at the plus end, promoting stability and growth.

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k-on and k-off Rates

The rates of addition (k-on) and removal (k-off) of subunits in filament dynamics.

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Equilibrium in Filaments

The state where the rates of growth and shrinkage are equal, leading to stable filament length.

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Actin Dynamics

The changes in behavior of actin filaments, including growth, shrinkage, and treadmilling.

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Plus End

The growing end of an actin filament where subunits are added.

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Minus End

The end of an actin filament where subunits are lost.

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Subunit Polymerization

The process where individual actin subunits (G-actin) join to form a filament (F-actin).

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Nucleotide Hydrolysis

The breakdown of nucleotides like ATP, which promotes dissociation of actin filaments.

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Listeria Movement

The locomotion of Listeria using actin treadmilling for motility inside host cells.

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Study Notes

Cytoskeletal Dynamics I

  • Cellular Dynamics Module: Focuses on cytoskeletal dynamics.

  • Cytoskeleton Components:

    • Actin filaments
    • Microtubules
    • Intermediate filaments
  • Mouse Fibroblast Cells: Used in studies of cytoskeletal dynamics.

  • Red Blood Cell Production: 100 million new red blood cells are produced every minute.

  • Learning Goals:

    • Understand shared and different properties of three kinds of cytoskeletons.
    • Grasp the common mechanisms of filament polymerization.
    • Understand the role of nucleotide hydrolysis in filament dynamics.
    • Understand treadmilling.
  • Lecture Outline:

    • What is the cytoskeleton?
    • Common properties of F-actin and microtubule dynamics.
    • Actin filament dynamics in vitro (treadmilling).
    • Microtubule dynamics in vitro (dynamic instability).
    • Accessory proteins controlling filament dynamics in vivo.
    • Drugs inhibiting cytoskeletal dynamics.
  • Relevant Chapters: MBoC (7th edition) Chapter 16.

Cytoskeletal Filaments

  • Three Types: Actin filaments, microtubules, and intermediate filaments.

  • Actin Filaments:

    • Helical polymers.
    • Involved in diverse processes (stress fibres, microvilli, muscle).
    • Important component of the cell cortex.
    • Crucial for cell shape change, division, and migration.
  • Microtubules:

    • Hollow cylinders made of tubulin.
    • Crucial for cell structure.
    • Involved in diverse functions like the mitotic spindle and cilia.
  • Intermediate Filaments:

    • Rope-like fibres.
    • Provide mechanical support.

Cytoskeleton and Disease

  • Actin Filaments: Myopathy, hearing loss, neuropathy, cancer, and kidney disease.

  • Microtubules: Cardiac disease, ciliopathies, neuropathy, and cancer.

  • Intermediate Filaments: Skin disorders, cataracts, neuropathy, and myopathy.

Actin Filaments - Details

  • Assembly Types: Helical polymers, stress fibres, microvilli, and muscle.

  • Monitoring Polymerization: Pyrene-conjugated G-actin and fluorescent reporters. The Pyrene-actin signal is 25-fold higher in the polymer form.

  • Sigmoidal Reaction Curve: Actin polymerization's characteristic curve. The curve exhibits a delay.

  • explained by nucleation (lag phase), elongation, and steady-state phases. -nucleation is rate limiting step

  • Critical Concentration: The free subunit concentration at steady-state.

  • Treadmilling: Actin subunits moving from the plus end to the minus end.

Microtubules - Details

  • Cryo-EM Reconstruction: Microtubule structures are reconstructed by cryo-electron microscopy.

  • Localization Diversity: Tubules show diverse patterns, including arrays and mitotic spindles.

Intermediate Filaments - Details

  • Hair Mutations: Mutations associated with curly hair (dog example).
  • Nuclear Lamina: Part of the nuclear envelope, similar to intermediate filament architecture.

Cytoskeleton Fundamentals

  • Polymer Structure: Cytoskeletons are polymers of smaller protein subunits.

  • Examples of Protein Subunits: Actin(G-actin), a-tubulin, β-tubulin, keratin, vimentin, Neurofilament protein, and Desmin.

iClicker Quiz

  • Plant Cells: Intermediate filaments are conspicuously absent.

Common Properties and Dynamics

  • Common Properties/Dynamics:

    • Subunit polymerization
    • Nucleotide hydrolysis
    • Plus-end is growing end
  • Dynamic Predictions: Critical concentration can predict various states such as growing, shrinking and treadmilling.

ATPase/GTPase Activity

  • Significance: Important for both polymerization and depolymerization.

Nucleotide Hydrolysis

  • Effect on Depolymerization: Promotes depolymerization as the resulting ADP/GDP form is energetically unstable.

Growing Filaments

  • GTP/ATP Caps: Growing filaments typically have a cap of GTP/ATP; this cap promotes continued growth.

Actin Treadmilling

  • Driving Force: Actin treadmilling drives the motility of Listeria.

  • Future Module: Listeria motility will be studied in a future module.

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