Topic 6 - Cytoskeleton Flashcards
What are the three types of cytoskeletal filaments?
- Intermediate filaments
- Microtubules
- Actin filaments
What is the purpose of the intermediate filaments?
Provide strength for the cell
What is the function of microtubules in the cell? (2)
Determine the positioning of organelles
Associate with motor proteins
What are the functions of actin filaments in the cell? (2)
Determine the shape of the cell surface
Responsible for whole cell locomotion
Cytoskeletal filaments would not be able to function without (broad term)
Accessory proteins
Intermediate filaments are only present in… (phyla, 3)
Vertebrates, nematodes, mollusks
In other words, “squishy” animals
Microtubules and actin filaments are only present in (taxonomic group)
Eukaryotes
Rank the general size in width of microtubules, intermediate filaments, and actin filaments from largest to smallest
Microtubules > Intermediate > Actin
Describe what cytoskeletal elements are important in mitosis and cytokinesis
Microtubules: form the mitotic spindle
Actin: forms a contractile ring to cleave cell during cytokinesis
When a cell is in motion (like a neutrophil chasing a bacterium), what part of the cytoskeleton is responsible for leading the movement of the cell?
Actin filaments
What element of a cell is responsible for a cell’s polarity?
The cytoskeleton (actin and microtubules, intermediate filaments are nonpolar)
Intermediate filaments adhere to (2) on the periphery of the cell
Desmosomes and hemidesmosomes
In non-polar cells, microtubules are arranged like…
Stars
In polar cells, microtubules are often arranged in this way…
Linear - helps create cell polarity
In polar cells (like epithelial cells), actin-dense areas on neighbouring cells are attached to each other via…
Adherens junctions
Why does the cell disassemble filaments and reassemble them as opposed to moving assembled filaments?
It is much harder to move an entire filament, but individual monomers can diffuse easily through the cell to produce continuous movement
Describe the + end of an actin or tubulin polymer
The fast-growing end
Describe the - end of an actin or tubulin polymer
The slow-growing end
What happens to a monomer of actin or tubulin when it goes from being free to being a part of a polymer?
It undergoes a conformational change
What are the 2 forms of actin and tubulin monomers?
T-form and D-form
When actin is in its T-form, it is bound to…
ATP
When tubulin is in its T-form, it is bound to…
GTP
When actin is in its D-form, it is bound to…
ADP
When tubulin is in its D-form, it is bound to…
GDP
An ATP/GTP cap is formed when…
Addition of subunits is faster than ATP or GTP hydrolysis
Monomers of actin and tubulin are added in the (1) form
T-form
Monomers of actin and tubulin dissociate from the polymer in the (1) form
D-form
Define the critical concentration of actin and tubulin monomers
The concentration at which the rate of polymerization is equal to the rate of depolymerization (addition = subtraction)
Define persistence length
Measures the stiffness of a filament: The minimum length of a filament at which random thermal fluctuations are likely to result in bending of the monomer
A small persistence length means…
The substance is flexible, even short lengths will bend in an energetically favourable way
What is the persistence length of actin?
20-40 um
In situations where actin is kept from bending, what is required to do this?
Accessory proteins
Which end of an actin monomer (+ or -) binds ATP?
- end
Describe what is happening to actin polymers during treadmilling
Net addition on one end of the polymer and net loss at the other end
What is profilin?
A protein which associates with actin to PROMOTE POLYMERIZATION
What is thymosin?
A protein which associates with actin monomers to INHIBIT POLYMERIZATION
Is it true that actin monomers can bind both profilin and thymosin?
No, can only bind one at a time
What is “nucleation”?
The initial process of actin polymerization in which 3 actin monomers come together to form the beginning of an actin filament
Actin filament nucleation occurs most commonly at or near…
The plasma membrane
Actin filament nucleation is regulated by…
External signals, i.e. cell signalling pathways
Actin filament nucleation is specifically regulated by (2)
ARP complex or formins
When active, ARP complexes…
Enhance polymerization of actin
Which end (+ or -) of the actin monomer does ARP bind to?
- end
When ARP binds to the - end of an actin filament, (1) is prevented
Depolymerization
when ARP binds to existing actin filaments, what happens?
Typically creates a branch 70 degrees off from the original branch - creates a meshlike 2D network of actin
Formins produce (branched/unbranched) actin filaments
Unbranched
Describe the “anatomy” of a formin
2 subunits (a dimer), each hold 1 actin monomer
Formins associate with the (+/-) end of an actin filament
+ end
How can the rate of polymerization or depolymerization of cytoskeletal filaments be altered?
With the addition of capping proteins
The two general forms of actin arrays in a cell are…
Bundles and networks
Actin bundles are made of (branched/unbranched) filaments
Unbranched
Actin networks are made of (branched/unbranched) filaments
Branched
How are actin bundles spaced out?
With the addition of alpha-actinin between the actin fibers to space them out
The presence of alpha-actinin in cytoskeletal arrangements allows this to come between actin fibres
Myosin-II
To prevent myosin-II from entering between contractile bundles, what protein holds actin filaments close to each other?
Fimbrin
Fimbrin is a (-imer)
Monomer
Alpha-actinin is a (-imer)
Dimer
A bundle of actin fibers will have this mixture of accessory proteins to properly space the fibers together
Trick question: there will only be either alpha-actinin or fimbrin, not both in a given bundle
What is filamin?
An accessory protein which associates with actin filaments to hold networks of actin filaments together
Filamin is a (-imer)
Dimer
Myosin is found in all members of this taxonomic grouping
All eukaryotes
In humans, how many distinct myosin families are there?
37
All (except 1) myosin type walk towards the (+/-) end of an actin filament
+ end
In order to produce “walking” movement, myosin uses…
ATP hydrolysis
Describe the anatomy of a myosin molecule
2 heavy chains (with globular heads, site of ATP hydrolysis)
2 light chains (associate with the head region)
The a-helix “tails” of myosin associate with other myosin dimers and form a bundle
What does it mean for a myosin bundle to be “bipolar”?
Myosin heads are oriented in opposite directions on either side of the “bare zone”
What causes rigor mortis?
In the absence of ATP upon death, the body tenses up due to the tight bond between actin and myosin
What stops rigor mortis?
Decomposition/breakdown
In the actin-myosin binding cycle, how often (in %) is myosin bound to actin?
Only 5% of the time, need to be able to synchronize this movement
How long (in nanometers) is each “step” of myosin-II?
8.5 nm
The power stroke of the actin-myosin binding cycle is caused by…
The removal of ADP from the N-terminal head group of myosin
What are the 3 sites of the troponin complex?
I = Inhibitory C = calcium binding T = tropomyosin binding
When there is no Ca++ present, is tropomyosin in or out of its groove?
Out of its groove - preventing the interaction of actin and myosin
When Ca++ is PRESENT, is tropomyosin in or out of its groove?
In its groove - out of the way, allows interaction of actin and myosin
Where are troponin complexes located in comparison to tropomyosin?
Troponin complexes at the end of tropomyosin molecules
How many Ca++ ions bind to the troponin complex?
4
What form of myosin carries cargo (like vesicles and organelles) along actin filaments?
Myosin V
Are there myosins in plants?
Yes! In all eukaryotes
The lever arm swing in Myosin V is this many nanometers long
30-40 nm
What are the 2 major groups of proteins that interact with actin filaments?
- Motor proteins
2. Accessory proteins
Tubulin is a (-imer)
Heterodimer: contains alpha and beta subunits
GTP can only be exchanged from (alpha/beta) subunits of microtubules
Beta
One strand of a mictotubule is called a…
Protofilament
The only exception where an alpha is not next to an alpha in a microtubule is…
At the “microtubule seam”
How many protofilaments are in a microtubule?
13
What protein is required for nucleation of microtubules?
gamma-tubulin
Nucleation of a microtubule starts at…
The microtubule-organizing center (MTOC) also known as the centrosome
What acts as the “template” which creates microtubules?
The gamma-tubulin ring complex
What makes up the gamma-tubulin ring complex?
7 gamma tubulins (each with 2 heads) and accessory proteins
What is a centriole?
A short cylinder of modified microtubules and accessory proteins
Microtubule complexes are very stable when arranged in a (1) configuration
Star/astral
In a case where microtubules are linear to make a polar cell, centrosomes will be…
At one end of the cell
Describe the MTOCs of plants and fungi
Have centrosomes (the complex that microtubules stem from) but do not have centrioles
If a cell is fragmented and microtubules are cut off from the MTOC, what will happen?
Microtubules will reform a MTOC, but it will not have any centrioles
Kinesin-13 and XMAP215 are examples of…
Microtubule binding-proteins, or MAPs
What end of a microtubule (+ or -) do dyneins move towards?
The - end
What are the largest and fastest motor proteins? (excluding the special myosin)
Dyenins: move at 14 um/s
What energy source do motor proteins use?
ATP (do NOT use GTP)
The “motor” of motor proteins is contained within…
The head region
When in dimer form, intermediate filaments are (polar/nonpolar)
Polar
When in tetramer form, intermediate filaments are (polar/nonpolar)
Nonpolar
How many tetramers associate to form a intermediate filament unit?
8 tetramers, can be joined to other groups end-to-end
What is the persistence length of intermediate filaments?
less than 1 um - very bendy!
Intermediate filaments are stabilized by…
Strong lateral hydrophobic interactions
Keratin, desmin, and neurofilaments are all kinds of…
Intermediate filaments
Keratin is found in this cell type
Epithelial cells
Neurofilaments are found in this cell type
Axons
Desmin is expressed in…
Muscle, all three types of it!
Do bacterial cells have a cytoskeleton?
Yes - they just don’t have exact same cytoskeletal elements, no intermediate filaments and have homologs for actin and myosin
What is special about fly embryonic development
Many nuclei form and then cellularization occurs
Tissue with many nuclei (like the fly embryo before cellularization) is called…
Synctium
Does the S phase take longer in humans or prokaryotes?
Longer in humans
What is the difference between mitosis and cytokinesis?
Mitosis = 2 nuclei Cytokinesis = 2 cells