11.2 Biochem (cell bio) Flashcards
Phases of the cell cycle
Mitosis (shortest)
Interphase: G1, S, G2
G0 is another phase, out of G1
G0 and G1 are of variable duration
Cyclins
Regulatory proteins that control cell cycle events.
They are phase specific (e.g only work in G1 phase)
Cyclins activate CDKs and make cyclin-CDK complexes
CDKs
Cyclin-dependent kinases
They are constitutive (expressed constantly) and inactive.
Must be activated by Cyclins.
Cyclin-CDK complexes
Must be both activated and inactivated for cell cycle to progress.
The complexes phosphorylate target proteins in order to drive the cell cycle.
(Kinases phosphorylate!)
What happens to cyclins once their phase-specific job is complete?
They are degraded by ubiquitin protein ligase
What proteins bind to and inactivate the cyclin-CDK complexes?
p21, p27, and p57
note: p53 controls the activation of p21
What cyclins are active at the G1 –> S transition? What TSGs are active there?
Cyclin D and Cyclin E
Rb and p53 (inhibit(!) progression thru the cycle at this point)
Where in the cell cycle is TSG p53 active?
G1 –> S and G2 –> mitosis
How does Cyclin D work?
Cyclin D binds and activates CDK-4.
The complex causes phosphorylation of Rb protein (kinases phosphorylate!)
Rb protein was bound to EF-2, but when it is phophorylated, it is released.
Since EF-2 isn’t bound, it can transcribe and synthesize all of the components needed for progression through the S phase
…which are:
Cyclin E
DNA polymerase
Thymidine kinase
DHF reductase.
How does Cyclin E work?
Cyclin E binds/activates CDK-2
This allows cell to progress from G1 –> S
What cyclins affect the G2 –> M transition of the cell cycle?
Cycin A and Cyclin B
What does Cyclin A do?
Cyclin A binds to CDK-2
This allows the cell to enter mitosis (prophase)
What does Cyclin B do?
Cyclin B binds to CDK-1
The Cyc B/CDK-1 complex is activated by CDC-25.
Once activated, the complex initiates breakdown of the nuclear lamins (nuclear envelope) so that mitosis can start.
What tumor suppressors are active in the cell cycle?
Rb and p53 normally inhibit G1 –> S
If mutated, there will be unrestrained growth.
Since they are TSGs, they need TWO hits to lose fn- having only once copy is good enough protection.
If Rb is mutated, what tumors result?
Retinoblastoma
Osteosarcoma
Permanent cells and the cell cycle
These cells stay in G0, and regenerate from stem cells.
Neurons, skeletal musc, cardiac musc, RBCs
If you give a pt a cancer drug, these cells will not be affected since they are not rapidly dividing.
Stable (quiescent) cells and the cell cycle
Are in G0, but enter into G1 and go thru the cell cycle when stimulated to do so.
Hepatocytes, lymphocytes
Labile cells and the cell cycle
Labile cells never go to G0, they are rapidly dividing and have a short G1 bc they go thru the cell cycle quickly.
Bone marrow cells, gut epithelium, skin, hair follicles.
Anti-cancer drugs affect these a lot.
What things regulate the cell cycle?
Cyclins
CDKs
TSGs
What happens in the RER?
Synthesis of secretory (exported) proteins
Addition of N-linked oligosaccharide to many proteins
Nissl bodies
Aka the RER of neurons
These synth enz (eg ChAT) and peptide neurotransmitters
What cells are rich in RER?
Mucus-secreting goblet cells of the small intestine
Ab-secreting plasma cells
Things that secrete a lot!
Free ribosomes
Not attached to a membrane.
Free ribosomes synth cytosolic and organellar proteins (for the mito, the nucleus)
Make things that stay inside the cell (RER makes things that will be secreted)
What happens in the SER?
Site of steroid synthesis
Detoxification of drugs and poisons
What kind of cells are rich in SER?
Liver hepatocytes*
Steroid-producing cells of the adrenal cortex
Things that detox or make steroids!
Lots of SER in hepatocytes of pts on lots of meds, bc need to detox a lot.
What are the two sides of the golgi called, and which way do they face?
cis-golgi is close to the ER (it receives stuff from the ER)
trans-golgi is closest to the plasma mbr, to sends stuff to the mbr or also to lysosomes
(the medial golgi is in between the cis and trans)
What AAs are modified by the golgi, and how?
Serine, Threonine, Asparagine
The golgi adds O-oligosaccharides to serine and threonine residues, and N-oligosaccharides to asparagine.
How does the golgi handle lysosomal proteins?
It adds mannose-6-P to them, which targets the proteins to the lysosome.
What happens if mannose-6-P is not added to lysosome proteins?
I-cell disease (inclusion cell dz)
Instead of being targeted to the lysosome, the lysosomal proteins just go through the default pathway, which is secretion outside of the cell. So, lysosomes can’t work since they don’t have the right proteins.
This is an inherited lysosomal storage disorder, often fatal in childhood. Results in: coarse facial features clouded corneas restricted joint mvmt high plasma lvls of lysosomal enz
What are the three fates of things that the golgi has packaged and released?
- Vesicle is targeted to they lysosome (bc it contains lysosomal proteins and is tagged w Mannose-6-P to go there)
- Goes into a constitutive transport vesicle and is excreted
- Goes into a secretory storage vesicle- is secreted when needed.
What things are sulfated by the golgi?
Sugars in proteoglycans
Selected tyrosine on proteins
The golgi used core proteins to assemble…
proteoglycans (and then it sulfates their sugars)
What are the 3 vesicular trafficking proteins?
COP-1 retrograde, golgi back to RER
COP-2 anterograde, RER –> cis-golgi
Clathrin- trans-golgi –> lysosomes
also endocytosis: plsm mbr –> endosomes
Receptor-mediated endocytosis
Cargo molecule binds to a cargo receptor on the pls mbr surface.
Adaptin helps Clathrin bind to the receptor, once enough bind, they are depressed inward (clathrin-coated pit), dynamin pinches them off
So a coated vesicle (clathrin-coated) is no in the cytoplasm.
Uncoating occurs (clathrin and adaptin can be reused), and the transport vesicle is naked.
Naked vesicle fuses w endosome/lysosome.
(really, fuses w endosome, hydrolytic enz enter, and this makes it a lysosome)
Lysosomes use hydrolytic enz to degrade things- this processes the endocytosed molecules so that they can be used by the cell.
T/F a clathrin-coated endocytosed vesicle fuses w the lysosome/endosome
False.
The vesicle must undergo uncoating before it can fuse.
Peroxisome
Mbr-enclosed organelle
Catabolizes long-chain FA and AAs
Changes long –> medium chain thru B-oxidation, and then they can go to the mito to be degraded.
Zellweger syndrome
Dysfnl peroxisome
Means that v long-chain FA will accumulate in blood- this causes neuro defects.
Proteasome
Barrel-shaped protein complex; it degrades damaged or unnecessary proteins that are tagged w ubiquitin
Nuclear localization sequence
Short 4-8 AA sequence rich in Arginine, Lysine, and Proline that targets the proteins to the nucleus (eg found on histones).
Nuclear pores recognize this sequence, and ATPase allows the protein to come in.
Very few things enter or exit the nucleus.
e.g. histones come in
mRNA and ribosomal subunits go out.
Chaperones
Assist in proper folding and txport of polypeptides across the ER, Golgi, beyond.
Some chaperones are synth’d constantly and are involved in normal intracellular protein trafficking.
Others are only made in times of stress (e.g. heat-shock proteins)
If the protein folding is not successful, chaperones facilitate degradation of the dmgd protein (often w ubiquitin)
Heat shock proteins
A type of chaperone protein that is induced by stress (in this case, heat).
hsp70, hsp90
They rescue shock-stressed proteins from misfolding.
Ubiquitin is also a hsp- it is added to proteins that misfold and need to be targeted for degredation by the ubiquitin-proteosome complex.
Microtubule
Cell transport structure
Cylinder md of helical array of polymerized a- and B-tubulin.
Each dimer has 2 GTP bound.
MTs are incorporated into flagella, cilia, and mitotic spindles; also involved in slow axoplasmic txport in neurons.
They grow slowly, but collapse quickly.
Molecular motor proteins
These drive up and down the MTs to transport cellular cargo on one end or the other.
Dynein does retrograde transport (+ to -)
Kinesin does anterograde txport (- to +)
Dynein = die! (move closer to the negative)
In neurons, which molecular motor protein carries NTs toward the synapse?
Kinesin
The MTs have their negative end at the cell body, and their positive end at the distal end of the axon.
So, NTs need to go from negative to positive, and kinesin does this.
Chediak-Higashi syndrome
Microtubule polymerization defect
Decreased fusion of phagosomes and lysosomes,
Decreased phagocytosis, bc phagocytes can’t mv
Px recurrent pyogenic infections, partial albinism, peripheral neuropathy.
List the drugs that act on MTs
Mebendazole/thiabendazole (anti-helminth) Griseofulvin (antifungal) Vincristin/Vinblastine (anti-cancer) Paclitaxel (anti-breast cancer) Colchicine (anti-gout)
Cilia structure
9+2 arrangement of MTs (9 MT doublets around outside, plus 2 single MTs in middle)
Each doublet has dynein ATPase that links all of the doublets and causes bending of the cilium by differential sliding of doublets
How do ciliated cells communicate so that the cilia on different cells beats in the same direction?
Gap junctions
Kartagener’s Syndrome
aka Primary ciliary dyskinesia
Dynein is defective, so cilia are immotile.
Causes male and female infertility (sperm, fallopian tube), bronchiectasis, and recurrent sinusitis bc bacteria and particles are not swept out.
A/w situs inversus
List the main cytoskeletal elements
Actin and myosin
Microtubules
Intermediate filaments
Where are actin and myosin found?
Microvilli
Cytokinesis
Adherens jns
Musc contraction
Where are Microtubules found?
Cilia Flagella Mitotic spindle Neurons (for axonal trafficking) Centrioles
List the intermediate filaments
IFs are a family of proteins w similar seq/structure. Most are cytoplasmic (except nuclear lamins are nuclear)
Vimentin Desmin Cytokeratin GFAP (glial fibrillary acid proteins) Neurofilaments L, M, H Peripherin Nuclear lamins A, B, C
Vimentin
Intermediate filament found in CT: in fibroblasts, leukocytes, endothelium
It supports cellular membranes and keeps certain organelles fixed within the cytoplasm
Desmin
Intermed Filament found in muscle cells- smooth, skel, and cardiac.
Cytokeratin
Intermed Filament found in epithelial cells
Keratin is in desmosomes and hemidesmosomes.
GFAP
Glial fibrillary acid proteins
Intermediate filaments found in astrocytes, schwann cells, and other neuroglia
Peripherin
Intermed filament found in neurons
Neurofilaments L, M, H
Intermediate filaments found in axons of neurons