Raiya Nerooban, Tobias Ezekial, Molly Bouw, Aditya Lumb, Chloe Jones, Teerka Baskaran, Jeffrey Pan, Brayden

Levillard

What are the functions of the cytoskeleton? (2 points)

The cytoskeleton permits common cellular functions such as signaling and vesicular transport to
occur and gives some cells unique properties, including cell motility. The cytoskeleton also
defines a cell's shape and the distribution of cellular contents.

Cytoskeletal proteins function similarly to other proteins. They bind to targets (often other
proteins) to form polymers. When the cytoskeletal proteins bind, they undergo conformational
changes. The function of these proteins is defined by the number and type of cytoskeletal
proteins that are bound.

What are the roles of the intermediate filaments, microtubules, and actin filaments within
cells? (3 points)

Intermediate Filaments are the strongest filaments, providing cells with the greatest mechanical
strength. There are 6 major classes of intermediate filaments, which vary in function and
distribution.

Protein Distribution Proposed Function

Acidic keratins Epithelial cells Tissue strength and integrity

Basic keratins Epithelial cells Tissue strength and integrity

Desmin, GFAP, vimentin, Muscle, glial cells, Sarcomere organization,

periphevin mesenchymal cells perphevin integrity
neurons

Neurofilaments Neurons Axon organization

Lamins Neurons Nuclear structure and

organization

Nestin Neurons Axon growth

Microtubules - The primary purpose of microtubules is to support trafficking within the cells.

Actin Filaments - The primary purpose of actin is to support cellular motility or other large-scale
movements like contraction.
 Raiya Nerooban, Tobias Ezekial, Molly Bouw, Aditya Lumb, Chloe Jones, Teerka Baskaran, Jeffrey Pan, Brayden Levillard

Identify three types of intermediate filaments, what are their functions and state any
differences between them (3 Points)

Lamins
- Found solely in the nucleus that forms the nuclear matrix, a dense network that protects
chromatin.
Desmins
- Don’t form long, thin, filamentous structures
- Responsible for muscle structural integrity
- Link different cellular structures together
Keratin
- Binds to desmosomes to form a complex
- Makes up hair, skin, and nails

Differences
● Lamin is found solely in the nuclear matrix, while desmins and keratins are found in the
cytoskeleton.
● Desmins don’t form long, thin, filamentous structures like lamins and keratin.

What is the purpose of microtubules (1 point), and how do they assemble (2 points)?
What are dynamic instability, catastrophe, capping proteins, and rescue (2 points)?

The primary purpose of microtubules is cellular trafficking. This traveling function can be
bidirectional along a single microtubule, where the cargo can attach or detach anywhere.

Dimers Form polymers: Microtubules assemble from bound alpha and beta tubulins to form a
dimer. Dimers form unstable polymers that can break down.
Polymer Growth: With six dimers, it’s known as a protofilament and becomes more stable. At
this point, it grows laterally or longitudinally.
Protofilament Tubes: Protofilaments form sheets and assemble into a tube of 13
protofilaments–the nucleation site for microtubule elongation.
Assembly / Disassembly: Dimers continue to assemble/disassemble at the ends of
microtubules. They will elongate if they are bound to GTP and disassemble if they are bound to
GDP.

Dynamic Instability: The cell’s ability to grow, shrink, and change direction very quickly in
response to changes in the cellular environment.

Catastrophe
When GTP is converted to GDP on the tubulin dimers at one end, they fall off, initiating a
catastrophe or the rapid depolymerization of tubulin dimers at the plus end, resulting in
microtubule shortening.
 Raiya Nerooban, Tobias Ezekial, Molly Bouw, Aditya Lumb, Chloe Jones, Teerka Baskaran, Jeffrey Pan, Brayden Levillard

Capping Proteins—Once the microtubule is at its desired length, the positive end can be
bound by microtubule capping proteins. These proteins add tremendous stability, keeping the
microtubule polymerized if the dimers are in the GDP-bound form.

Rescue —If catastrophe occurs, it can be rescued if enough GTP-bound dimers are present,
but it can also occur in the presence of other proteins.

Describe the stages of actin polymerization and explain treadmilling and how it is
regulated (5 points)

1. Nucleation - two actin monomers can dimerize, but nucleation occurs when a third actin
monomer binds to the dimer to form a “nucleus trimer.” This forms the core on which the
rest of the actin filament can form. This is structurally simpler than MTOC, but it serves
the same purpose.

2. Elongation - growth occurs from both directions, starting at the nucleus, but it is
favoured at the plus end. Actin polymerization is also dynamic, with actin monomers
added and removed. If the balance favours adding monomers, then the filament
elongates.

3. Steady State - The rate of assembly equals the rate of disassembly and elongation
ceases.

Treadmilling – favoured addition of monomers to one end with the same rate of monomer
removal from the other. This effectively keeps the actin filament the same length but can result
in the filament moving within the cell. Treadmilling allows cells to rapidly adjust the actin
cytoskeleton, much faster than the intermediate filaments, which require phosphorylation for
disassembly.

Regulation
Treadmilling is regulated by the concentration of ATP-bound actin monomers compared to the
concentration of ADP-bound monomers. The critical concentration for polymerization of
ATP-bound monomers is lower at the plus side, meaning at the right concentrations, ATP-bound
monomers will be added to the plus side. In contrast, ADP-bound monomers are removed from
the minus side at the same rate. However, if ATP-bound monomer concentrations increase at
the minus end, monomers will also be added to the minus side.
 Raiya Nerooban, Tobias Ezekial, Molly Bouw, Aditya Lumb, Chloe Jones, Teerka Baskaran, Jeffrey Pan, Brayden Levillard

What are microtubule based motor proteins (0.5 points). Describe the two most common
motor proteins (2 points). Explain the process of the walking of motor proteins (2.5
points)

Motor proteins control trafficking and support the movement of cargo within the cell, while
microtubules control where molecules can go.

1. Kinesin
- Move along microtubules toward the positive end
2. Dynein
- Move along microtubules toward the minus end

Heads contain microtubule-binding domains, while tails bind to the cargo that needs to be
trafficked. Both have two heads that walk along the microtubules, which consume ATP.
Walking/movement requires energy in the form of ATP.

Walking of Motor Proteins:

- Step 1: head one is bound to the microtubule, and head two is bound to ATP.
- Step 2: walking movement is initiated by ATP binding to head 1, causing a
conformational change that includes head 2 swinging around.
- Step 3: head 2 binds to the microtubule and releases ADP.
- Step 4: the ATP at head 1 undergoes hydrolysis, so it is ADP bound to head 1, causing
release from the microtubule.
- Step 5: The entire process is then repeated, but this time, ATP is binding to head 2,
causing head 1 to swing around.

Actin binding proteins play critical roles in organization and regulating actin filaments.
Hypothesize the consequences of dysfunctional or absent nucleating proteins, capping
proteins, membrane anchors and actin-binding motor proteins. Which do you think
would have the greatest impacts on cellular function? (5 points)

Nucleating proteins Bind to actin polymers to increase their stability and allow for the growth of
a new branch. Dysfunctional or absent nucleating proteins could lead to less stable actin protein
networks and less complex/branched networks. The nucleating protein is involved with actin
polymerization, and the lack of actin nucleators is linked to immunodeficiency (Moulding et al.,
2013).

Membrane anchors Link actin filaments to nonactin structural proteins similar to those integral
to the plasma membrane, such as integrins. Without membrane anchors, the cell’s structure
would not be as stable. If actin filaments are not anchored to the membrane, there is no tension,
and the cellular shape/structure can be altered (Wang et al., 2019).
 Raiya Nerooban, Tobias Ezekial, Molly Bouw, Aditya Lumb, Chloe Jones, Teerka Baskaran, Jeffrey Pan, Brayden Levillard

Actin-binding motor proteins bind to the actin filament and allow movement. Without these
motor proteins, cells wouldn’t be able to move or contract because they would lack the proteins
needed for movement.

Capping proteins bind to the plus or minus end and function to stabilize the polymer to prevent
disassembly or further assembly. If there were dysfunctional or absent capping proteins,
filament assembly would increase, which in vitro experiments have shown to decrease
lamellipodia and increase filopodia formation. Lamellipodia are in charge of cell movement,
meaning that with fewer lamellipodia, the cells would no longer be able to move, halting most
bodily functions (Management, 2023). GREATEST IMPACT: A decrease in capping proteins
would eventually lead to no cell movement, preventing large-scale contractions like
muscle contraction and causing cellular death. Additionally, it would prevent the cells
from regulating their actin networks.
 Raiya Nerooban, Tobias Ezekial, Molly Bouw, Aditya Lumb, Chloe Jones, Teerka Baskaran, Jeffrey Pan, Brayden Levillard

References
Management. (2023, November 30). What is capping protein? - Mechanobiology Institute,

National University of Singapore.

https://www.mbi.nus.edu.sg/mbinfo/what-is-capping-protein/

Moulding, D. A., Record, J., Malinova, D., & Thrasher, A. J. (2013). Actin cytoskeletal defects in

immunodeficiency. Immunological Reviews, 256(1), 282.

https://doi.org/10.1111/imr.12114