Cell And Molec Test 3 Flashcards
What are the fibers of the cytoskeleton made out of? (3)
- microfilaments
- intermediate filaments
-microtubule
What are the differences between micro filaments, intermediate filaments, and microtubules?
- microfilaments are the smallest, most simple and made of actin ~7nm
- intermediate has 9 types and is a complete structure ~10nm
-microtubules are the largest, they are a hollow tube made out of tubulin(?) ~25nm
Do prokaryotes have a cytoskeleton? Do eukaryotes?
Only eukaryotes have a cytoskeleton, prokaryotes do not
What are the 4 techniques for visualizing the cytoskeleton?
- fluorescence microscopy on fixed specimens
- live cell fluorescence microscopy
- computer-enhanced digital video microscopy
- electron microscopy
Describe how Fluorescence microscopy on fixed specimens works
Fluorescence compounds directly bind to the cytoskeletal proteins, or antibodies are used to indirectly label cytoskeleton proteins in chemically preserved cells, causing them to flow in the fluorescence microscope
Describe how live cell fluorescence microscopy works
Fluorescence versions of cytoskeletal proteins are made and introduced into living cells. Fluorescence microscopy and video of digital cameras are used to cure the proteins as they function in cells
Describe how Computer enhanced digital video microscopy works
High resolution images from a video or digital camera attached to a microscope are computer processed to increase contrast and remove background features that obscure the image
Describe electron microscopy
Electron microscopy can resolve individual filaments prepared by thin section, quick freeze dry etch, or direct mount techniques
What is the largest of the cytoskeletal components of a cell
Microtubules
What are the functions of microtubules?
- move chromosomes (main function)
- flagella movement
- “highways” for vesicles
What 2 types of microtubules are responsible for the many functions in the cell?
-cytoplasmic microtubules
-axonemal microtubules
What are the functions of cytoplasmic microtubules
-maintaining axons (axons are the long structures in nerve cells)
-formation of mitotic and meiotic spindles
-maintaining or altering cell shape
-placement and movement of vesicles
What are axonemal microtubules?
they include the organized and stable microtubules found in structures such as : cilia, flagella, and basal bodies (cilia and flagella attach to)
What is the axoneme?
A highly ordered bundle of microtubules
Describe what is shown in the graph (in my notes) (LAG PHASE ELONGATION PHASE AND PLATEU PHASE
The first part of the graph (low/start) is the lag phase which is the period of nucleation
Then, during the elongation phase, (middle part) microtubules grow rapidly causing the concentration of tubulin subunits in the solution to decline when this concentration is low enough to limit further assembly, the plateau phase (high point flat at end) is reached during which subunits are added and removed from microtubules are equal rates
What is critical concentration?
Concentration at which the rate of assembly of cytoskeletal protein subunits into a polymer is exactly balanced with the rate of disassembly
What is the goal of the The microtubule organizing center (MTOC) (idk how to ask this question)
It anchors the negative end of the microtubule so it’s bound to the MTOC so it can’t shrink or grow. It can only shrink/grow at the positive end
What happens to the microtubule when there is a high tubulin concentration
The microtubule grows, GTP-tubulin subunits are added
What happens to the microtubule when the tubulin concentration is low?
The microtubule shrinks, GTP are hydrolyzed
At what end do microtubules grow/shrink
They grow and shrink at the positive end, they anchor at the negative
Which end of the microtubule is more dynamic? Why?
Positive end because that is where GTP is added and taken away (depending on the tubulin concentration) aka where it grows and shrinks
What happens in the MTOC to the microtubules when tubulin concentration is high?
GTP-tubulin subunits are added
What happens in the MTOC to the microtubules when there is low concentration of tubulin?
Catastrophe. GTP are hydrolyzed (therefore depleting the GTP cap)
Microtubules are regulated by microtubule binding proteins. Why might they use ATP?
-drive vesicle/organelle transport
-generate sliding forces between microtubules
What are the functions of microtubule-binding proteins?
-drive vesicle or organelle transport (using atp)
-generate sliding forces between microtubules (using atp)
-regulate microtubule structure
Cells regulate _______ with great precision
Microtubules
What are microtubule-stabilizing/building proteins? (MAPS)
microtubule-associated proteins that bind at regular intervals along a microtubule wall. this allows for interaction with other cellular structures and filaments
What are the 2 microtubule stabilizing/bundling proteins (MAPs)?
- A MAP called Tau
- MAP2
What is the function of A MAP (Tau)?
They cause microtubules to form right bundles in axons
What is the function of MAP2?
they promote the formation of looser bundles in dendrites
What are dendrites?
short branched extensions of nerve cells (neurons) that receive signals from other neurons
Describe what the 2 regions of MAPs such as Tau and MAP2 each do
One region binds to the microtubule wall, and the other extends at right angles to allow for interaction with other proteins
What controls the spacing of the microtubules in a bundle?
The length of the extended “arm” of the MAP
What are +-TIP proteins?
- (+- end tubulin interacting proteins) are stabilizing proteins that “capture” and protect the growing positive end
-they decrease the likelihood that microtubules will undergo catastrophic subunit loss
What are 3 microtubule destabilizing/severing proteins?
- Stathmin/Op18
-Catastrophins
-proteins such as Katanins
How does Stathmin (or OP18) destabilize/sever microtubules?
-it binds to tubulin heterodimers and prevents their polymerization
How do catastrophins destabilize/sever microtubules?
at the end, they act on the microtubules and promote the peeling of du units from the ends
How do katanins (proteins) destabilize/sever microtubules
They sever the microtubules
What is “The Mesh” ?
-a network of microtubule connectors that stabilize individual kinetochore fibers of the mitotic spindle where the chromosome attaches
- yellow “lines” in the given photo
Each mesh structure holds 2 microtubules together, why?
Because the clathrin is a part of the mesh structure (in its turmeric state)
What is the smallest of the cytoskeletal filaments?
Microfilaments
Microfilaments have many roles, what do they include? What is the best known role they carry out?
- BEST KNOWN : their role in muscle contraction
also known for : - cell migration
- ameoboid movement
- cytoplasmic streaming
- development and maintenance of cell shape (they push out the edges of the cell)
- structural core of microvilli
What is actin?
- a very abundant protein in the cell, it is a building block for all microfilaments
Once actin is synthesized, it is folded into a globular shaped molecule. What does this allow it to do?
It allows the actin to bind to ATP or ADP
Since microfilaments do not anchor themselves at one end, where do they shrink/grow
There is general shrinkage at the negative end and general growth at the positive end
When the myosin fragment S1 binds to the actin in microfilaments, a “pattern” is created - describe why this is
The myosin segment binds to the actin and creates an “arrow” pattern, where the arrow will point to the negative end creating the “pointed end” and the other end is referred to as the “barbed end”
Which is not dynamic: microtubules or microfilaments
Microtubules are more dynamic
I’m terms of length, which is more stable: microtubules or microfilaments?
Microfilaments are more stable in terms of length
Explain “tredmilling” in terms of microfilaments.
-since they gain G-actin monomers at the positive end while simultaneously they lose monomers at the negative end, the net change in length is zero.
-the actin monomers start at plus end and are displaced progressively more towards the negative end until depolymerization causing it to detach
When is the ONLY time “tredmilling” in microfilaments occurs?
In a test tube/in vitro
What are 3 specific drugs that affect polymerization of microfilaments?
- Cytochalasins
- Latrunculin A
- Phalloidin
How do Cytochalasins affect the polymerization of microfilaments?
- the prevent the addition of new monomers to existing microfilaments
- cytochalasins are fungal metabolites
How does Latrunculin A affect the polymerization of microfilaments?
- hides actin monomers and prevents their addition to microfilaments
- a toxin
How do Phallodin affect the polymerization of microfilaments?
- stabilizes the microfilaments and prevents their depolymerization
- can alter and even kill the cell
Cells can precisely control where actin assembles and the structure of the resulting network, how?
- they use a variety of actin-binding proteins in order to control the actin
- control occurs at the nucleation, elongation and severing of microfilaments
What are the most stable, and least soluble cytoskeletal components?
- intermediate filaments are both the most stable and least soluble
What is an example of an intermediate filament (very abundant)
- keratin, an important component of structures that grow from skin in animals; giving structure to the skin
How many different classes of intermediate filament proteins are there?
- 6
Which of the 6 classes of intermediate filament proteins is the most stable?
- class 5 (V)
Class I intermediate filaments are _____
- acidic keratins
Class II of intermediate filament proteins ___
- are basic or neutral keratins
What 2 classes make up tonofilaments? Where are the tonofilaments found?
-protien classes I and II
- found in epithelial surfaces covering the body and lining it’s cavities
Class III of intermediate filament proteins include ___
- vimentin (connective tissue), desmin (muscle cells) and glial fibrillary (GFA)
Class 5 (V) includes ____
- nuclear Kamins A, B, and C, that form a network along the inner surface of the nuclear membrane
Class 6 (VI) of intermediate filament proteins ____
- neurofilaments in the nerve cells of embryos are made of nestin
What is intermediate filament testing?
-animal cells can be distinguished based off of what types of intermediate filaments they contain
What are nucleotide triphosphates (NTP)?
- (rna) an umbrella term for : ATP, GTP, UTP, and CTP
What are deoxynucleotide triphosphates?
- (dna) dATP, dGTP, dCTP, dTTP
What is an allosteric site?
- a binding site distinctively different from the active site, that allows molecules to either activate or inhibit enzyme activity
What is a holoenzyme?
A complete enzyme including a cofactor, the enzyme, and once the substrate binds it is functional
Explain how the intermediate filament is assembled in vitro
2 intermediate filament proteins go through coiling, creating a coiled-coil dimer of 2 intertwined polypeptides.
Then, the coiled-coils line up (lateral alignment) creating a tetrameric protofilament of 2 aligned coiled-coil dimers.
They are then assembled in a staggered pattern (like bricks) into a long rope-like filament. The final structure consists of 8 protofilaments (16 coiled dimers) in staggered overlaps.
Which of the following is both flexible and can withstand tensile forces : microtubules, microfilaments, or intermediate filaments
- intermediate filaments
How do the microtubules and microfilaments work to support the cytoskeleton?
The microtubules resist bending when a cell is compressed while the microfilaments serve as contractile elements that generate tension
The nuclear lamina (on the inner surface of the nuclear envelope) disassembles when? When do they reassemble?
They disassemble at the onset of mitosis, and then reassemble afterward
Are intermediate filaments considered a static structure?
No. They are not static, they are dynamically transported and remodeled
What are Plakins?
- linker Proteins
What role does Plakins have in the cytoskeletal structure?
- the Plakins proteins connect the intermediate filaments, microfilaments, and microtubules
What is an example of a Plakin (name of it)
-plectin
Where is plectin found in the cytoskeleton?
At sites where intermediate filaments connect to microfilaments and microtubules
What are the 2 microfilament associated (actin binding) proteins? (Motor molecules)
- myosin I - monomer
- myosin II - filament (specialized)