Cell Biology Final Review

Chapter 15 Signal Transduction 1) Endocrine, paracrine , autocrine call forise, and cell-cell contact (Fig. 15-2). Endocrine signaling is eagle-eyed distance signaling. An mannequin would be pancreatic cells secreting insulin. Paracrine signaling is for close proximity. An example would be a nerve cell releasing neurotransmitters. In autocrine signaling the cell that produces the ligand also contains the receptor for that ligand. This is how cancer cells work. In signaling by plasma membrane attached proteins, the target cell does something in receipt to direct contact from the signaling cell. ) List examples of 1) steroid hormones and 2) amino covering breaker derivatives that act as ligands. What be the catecholamines, and which amino acid ar they derived from? steroid hormones moor cytosolic receptors. They include cortisol, progesterone, estradiol, testosterone, thyroxine and retinoic acid. Steroid receptor complexes increase or decrease the transcription rates of cert ain genes. Dopamine, norepinephrine, epinephrine, serotonin and histamine argon ligands that atomic number 18 derived from amino acids. Catecholamines are ligands derived from the amino acid tyrosine. 3) Whats an agonist?Whats an obstructor? A doctor prescribes isoproterenol to his patient why? Why not epinephrine? Another patient receives alprenolol why? (See page 629 and Fig. 15-5). Agonist= structural analog, opposer=inhibitor. Isoproterenol has lower Kd ( exalteder affinity) than epinephrine, and will inc. smooth heart vigor contraction. Alprenol is an antagonist (beta blocker) and slows heart contractions 4) What are the five kinds of second messengers we draw in lecture. (Fig. 15-9 shows only four) camping, cGMP, DAG, IP3 and Ca2+ 5) What are GTP- compeling (switch) proteins?When are they on? When are they tally? (Fig 15-8). GEFs help annul them on. GAPs help turn them off. Are on when bind GTP, and off when bind GDP. Ex Ras, Ran, trimeric G proteins 6) What are kin ases versus phosphatases? Kinases phosphory late, Phosphatases dephosphorylate 7) What are the briny features of a G-protein coupled receptor (GPCR, Figs. 15-10, 15-12, and 15-13)? What is epinephrine and what kinds of receptors does it bind on what cells to figure of speech what responses? GPCRs= seven-pass receptors with amino terminus outside cell and carboxy terminus inside cell. epinephrin fired when glucose needed quickly (inc. animal starcholysis and lipolysis) binds to GPCR receptor , which activates a G protein (switch protein), which activates an effecter protein (adenylyl cyclase) producing d head NOTE bivouacking does not involve RTK (tyrosine), simply uses Ser/Thr kinases PDE degrades cAMP 8) come upon the terce G proteins ? , ? , and ?. Which one binds GTP/GDP (hint for read/write head 6 above). G? binds GDP, is tethered to national leaflet of plasma membrane, nevertheless dissociates from ? and ? to activate effector protein (adenylyl cyclase). G? and G? n ever separatedAre tethered to inner leaflet and work as a unit. 9) Describe FRET (Fig. 15-14). 10) What is adenylyl cyclase? Figs 15-21 and 15-22. What does it do? How is adenylyl cyclase positively and negatively controlled? Positive epinephrine binds ? adrenergic receptors to activate Gs, actvating adenylyl cyclase ostracise PGE binds to ? adrenergic receptors to activate Gi which accordingly inhibits adenylyl cyclase. 11) The complete Fight or Flight road map. YIKES What happens when cAMP rises? What happens when cAMP drops? 12) T/F secondly messengers are long-lived in their signaling effectiveness?How is cAMP vitiated? PDE 13) What is cAMP-dependent protein kinase and how does it work? (Figs. 15-23 and 16-31) Do not involve tyrosine kinases (RTK), that use Ser/Thr kinases (binding of cAMP releases catalytic sites) 14) What do we mean by amplification in signal transduction? Fig. 15-26. So many steps involved in signal transduction b/c youre amplifying signal at every step unbendable response 15) PIP2, DAG, IP3, and the release of Calcium from the endoplasmic reticulum. (Fig. 15-30). Each PI kinase phosphorylates inositol ring PI PIP PIP2, and cleavage of PIP2 by Phospholipase C yields DAG and IP3.Phospholipase C is activated by a hormone binding to GPCRs and activation of G proteins. IP3 releases Ca2+ back into cytosol to transduce a signal (Ca2+ binds to PKC which binds to DAG phosphorylates substrates). Ca2+ pumps normally pump Ca2+ (from cytosol) into ER or out to exterior yet IP3 causes ion channels to untied and release Ca2+ into cytosol. at one time Ca2+ released, it positively feeds back on channels to allow to a greater extent than Ca2+ to flow out. But once Ca2+ becomes depleted from ER and at high conc. in cytosol, it inhibits channels. ALSO once Ca2+ rises in cytosol, acts as a second messenger to trigger insulin release 6) Calmodulin. Activated by binding of 4 Ca2+ molecules, it then activates PDE (to degrade cAMP), animal starch ph osphorylase kinase GPK (to break down glucose, activates this path without cAMP ), other protein kinases, and Nitric Oxide (NO) synthase (involved in acetylcholine relaxation of smooth muscle in conjunction with cGMP) 17) How are blood vessels dilated by acetylcholine (Fig. 15-31)? BTW, what does Viagra do? Acetylcholine binds acetylcholine GPCR, which activates phospholipase C, which makes IP3, which binds to Ca2+ (leading it to inc. n cytosol), and Ca2+ binds calmodulin, which activates NO synthase that produces NO. The NO is then released by paracrine signaling into muscle cells and binds NO receptor that converts GTP to cGMP, which activates protein kinase G relaxation of muscle cell and by endocrine signaling causes blood vessel dilation. VIAGRA blocks degradation of cGMP by PDE (may cause blindness b/c rod cells kept fan out by cGMP) 18) Beta arrestin in receptor desensitization. If receptor constantly candid to epinephrine, may itself become phosphorylated by PKAblocking t ransducing signal, downregulating ALL GPCRs.Once ? -adrenergic receptor is phosphorylated by BARK (? -adrenergic receptor kinase) ? -arrestin binds the receptor to block its activation of Gs, as well as promotes formation of *clathrin-coated vesicles for endocytosis of the butt on receptor (to deplete surface receptors) CHAPTER 16 Signal Transduction and Gene Expression 1) List some(prenominal) ligands that bind to sense organ Tyrosine Kinases (RTKs). * Nerve ontogenesis factor (NGF), home platelet derived growth factor (PDGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), insulin, and moreList some(prenominal) kinds of general responses that could occur. What characteristics are different between RTKs and GPCRs (i. e. their protein structure and function)? * RTK= receptor tyrosine kinases halte components extracellular ligand-binding site, a single hydrophobic trans-membrane ? helix, and a cytosolic catalytic domain. Ligand binding causes a conformational cha nge that promotes formation of a functional dimeric receptor, bringing unneurotic cardinal poorly active kinases that then phosphorylate each other on a tyrosine resi due in the activation lip.Phosphorylation causes the lip to move out of the catalytic site, thus allowing ATP or a protein substrate to bind. The activated kinase then phosphorylates other tyrosine residues in the receptors cytosolic domain. The resulting phophotyrosines function as go in sites for various signal-transduction proteins. * GPCR= G-protein coupled receptors. Binding of ligand triggers the exchange of GTP for GDP on the G? subunit and dissociation of G? GTP from the G complex, and G? subunit transduces the signal, but in yeast pheromone receptors its the G complex. G unctions by triggering a kinase cascade (similar to the one for Ras). Its proteins are involved in mating-specific cellular responses. 2) What are adaptor proteins? Specifically what are SH2, SH3, and PTB domains and how do they work. (Figs . 16-19, 16-20) No intrinsic enzyme body process have docking sites for other effector proteins, such as SH2, SH3, or PTB domain (Phospho-Tyrosine Binding). These docking proteins pass the signal onto Ras. 3) Growth FactorRTKGRB2SOSRasRafMEKMAPdifferential gene aspect for cell division or specific cell type differentiation.What happens at each step? (Figs. 16-21, 16-22, 16-25, 16-27). Why so many steps (see question 16 in the previous section)? Sev gene regulates R7 development and in mutants R7 is abstracted cell differentiates into a cone instead and flies now cranky to UV light. The Sev gene product is RTK and Boss (in R8 cells) is the ligand for this RTK. Once Boss binds/activates Sev RTK it causes GRB2 (with SH subunits) to bind receptor, leading to relocation of SOS (the GEF for Ras) from the cytosol to the membrane where Ras-GDP resides and it activates it ) Phosphatidylinositol 3,4,5 tri-phosphate and Protein Kinase B (Fig. 16-29 and 16-30). PI-3,4,5-triphosphate ( make by phosphorylation of PIP2 at 3 OH by PI-3 Kinase) is docking site for Protein Kinase B. PKB bound to PI 3,4,5-triphosphate and PDK1 (also bound to PI 3,4,5triphosphate) dif primer coat into membrane and PDK1 phosphorylates/activates PKB Ras-independent insulin signaling 5) Insulin versus glucagon. (Slide from lecture shows a delay comparing and contrasting the two ligands and their effects on serum glucose. ) Insulin synthesized in ? ells and when theres high blood glucose activate GLUT4 (glucose transporter) and inc activity of glycogen synthase removal of glucose from blood and its storage as glycogen. Glucagon reacts to a decrease in blood glucose, stimulating release of glucagon, activating adenylyl cyclase, activating glycogen phosphorylase and inhibiting glycogen synthase degradation of glycogen and release of glucose into blood. Chapter 20 The Cell Cycle 1) Who were the three Nobel Prize winners in Physiology and Medicine for 2001? * Leland Hartwell * Tim Hunt * Paul Nurse ) polish up the gross morphological events of prophase, metaphase, anaphase, and telophase. * Prophase * Chromosomes condense to the 30nm solenoid fiber * Chromatids remain attached at the centromeres and the spindle forms * The atomic gasbag disassembles in most eukaryotic cells (called ease up mitosis). Yeasts and other fungi have a closed mitosis * ER and Golgi turn into vesicles * Metaphase Condensed chromosomes align in a straight line that is referred to as the metaphase plate * Anaphase * Sister chromatids separate from each other The spindle is critical for chromatid endeavor to opposite poles * Molecular motors generate force and movement * Telophase * Beginning of the future(a) interphase * Chromosomes stupefy to decondense * The nuclear envelop and the nucleolus begin to reassemble * Cytokinesis * cytoplasm discriminates * Golgi and ER reform from vesicle fusion 3) Figure 20-2 is a practised summary. 4) Three major classes of Cdk/cyclin complexes Where they work in the cell speech rhythm, and what do they do. What are the three critical steps in the cell regular recurrence? * G1 cyclin-CDKs convey when growth factors (EGF, PDGF, NGF, etc) signal the cells to divide * Phosphorylates the retinoblastoma protein in mammal cells * S-phase cyclin-CDKs * Form during G1, but are held silent by an inhibitor * The inhibitor is destroyed by ubiquitin-mediated proteolysis. Then, the cell progresses into S-phase * Mitotic cyclin-CDKs * too called MPF ( festering/mitosis promoting factor) * Form in S-phase and G2, but are held silent until late G2 * Once activated, mitotic Cdk-complexes initiate mitosis * Chromosome condensation Nuclear envelope disassembles and the spindle forms * Chromosomes align on the metaphase plate * ER and Golgi turn into vesicles * Partially activates anaphase promoting complex (APC) 5) Classic experiments * What happens when you fuse a G1 cell to a M-phase cell? Fig. 20-3 * Interphase cells advance prematuring into M-phas e * Now we bonk that the diffusible regulators are the mitotic Cdk-complexes (MPF) * What happens when you fuse a G1 cell to a S-phase cell? * G1 nuclei begin to replicate their deoxyribonucleic acid prematurely Used 3H-thymidine incorporation and autoradiography to visualize deoxyribonucleic acid synthesis * Now know that diffusible S-phase Cdk-complexes activated the pre- repercussion complexes on deoxyribonucleic acid origins of replication in the G1 nuclei * What happens when you fuse a G2 cell to a S-phase cell? * Re-replication of G2 deoxyribonucleic acid does not occur * Once DNA is replicated, it cannot be re-replicated in that analogous cycle * Whats the diffusible regulator in the first experiment? MPF 6) What two species of yeast were used to decipher the genetics of the cell ycle? Whats a closed mitosis versus and open mitosis? * Budding and fission yeast * In open mitosis, the nuclear envelope disassembles during mitosis. In closed mitosis, the nuclear envelop doe s not disassemble. 7) What is cloning by complementation? (Fig. 20-4). This is the same as functional complementation. * Many cdc mutations identified are temperature sensitive * Grow and divide at permissive temperatures * Fail to divide at non-permissive temperatures * We can select cDNAs by functional complementation ) What is MPF (what two proteins make up MPF) and where did its name come from (i. e. what organism and cell type)? (Figs. 20-5 and 20-6) * MPF is the maturation promoting factor. It is comprised of Cdk1-Cyclin B * The name came from studying frog oocyte maturation in vitro 9) The piece of land to MPF conclusion What is Anaphase Promoting Complex (APC)? What activates APC? What does APC then do and how does it do it? Fig. 20-10 * MPF is a kinase that phosphorylates many different substrates to initiate mitotic events * To exit mitosis, MPF must be destroyed Destruction of MPF depends on the destruction of Cyclin B * Destruction of Cyclin B is via the ubiquitin path way * Ubiquitin is covalently linked to lysines behind the destruction box * Cyclin B without the destruction box will not be destroyed * APC destroys MPF, but APC was actually activated earlier at anaphase by MPF 10) You have to know Fig. 20-13 and 20-14 11) Molecular events at the onset of mitosis a) Nuclear envelope dismantlement what are the lamin proteins, how do they disassemble, and where do they go when they disassemble? (Figs. 0-16, 20-17) * The nuclear lamina supports the nuclear envelope. It is found on the underside of the inner envelope membrane. * The nuclear lamina is made of three lamin proteins A, B and C * All three lamin protein form coiled-coil dimers * both dimers form a tetramer with head-to-head or tail-to-tail orientations * MPF phosphorylation of Ser residues causes disassembly * A and C diffuse into the cytoplasm. B remains bound to the membranes that form vesicles during mitosis b) Condensation of chromatin granule what are the SMC protein (condensins)? * SMC proteins in yeast Structural maintenance of chromosomes (SMC) * Large proteins that form coiled-coils * ATPase activity in their C-terminus * Function in the normal segregation of chromatids * SMC proteins in frogs * Part of a complex called condensin that becomes phosphorylated at the onset of mitosis * Condensins bind DNA and wind it into supercoils with ATP hydrolysis * Several condensins bind along the lenth of the chromosomes to form coiled-coils to cluster the DNA c) Spindle assembly due to MPF phosphorylation of microtubule-associated proteins d) ER and Golgi vesiculation due to direct MPF phosphorylationWhat are the cohesins what do they do and what regulates their activity? How does APC extend a role in this regulation? (Fig. 20-21, 20-22) * Cohesins hold sister chromatids together * Separation of chromatids is not dependent on MPF destruction * Cohesin function is regulate by an anaphase inhibitor called securing. This inhibitor is a target for APC ubiquitination 12) Yeast cell cycle (Figs. 20-29 and 20-28) * Sic1 is the S-phase inhibitor destroyed by ubiquitination * E3 for the ubiquitination is cdc34 * The ubiquitination complex is called SCF Once Sic1 is destroyed, Cdc28/Clb5 + 6 phosphorylate substrates to initiate DNA replication * G1 cyclin-Cdc28 phosphorylated Sic1, enabling its recognition and ubiquitination by Cdc34 and SCF * Cln1 and Cln2 arise early on in the cell * Cdc28 is only in yeast * Clb5 and Clb6 arise late in G1. They are called S-phae cyclins. They are rapidly turned on by the destruction of Sic1 13) Why chromosomes replicate only once per cell cycle (Fig. 20-30) * Protein degradation makes cell cycle progression move forward, not upset * Sic1 is destroyed at G1 to S-phase renewing Anaphase inhibitory (securing) is destroyed at the metaphase to anaphase transition * Cyclin B is destroyed at the mitosis to G1 transition 14) Early and late mammalian response genes. 15) mammalian cell cycle (Fig. 20-32) * Growth factor h ormones are called mitogens * The absence of mitogens cases cells to arrest in G1 or G0 * If mitogens are added, cells advance sometime(prenominal) the restriction point and are committed to S-phase and mitosis * Mammalian cells have several Cdks * Cdk 1, 2, 4, and 6 are used for the cell cycle * Cdk 1 complements Cdc2 * Mammalian cells also have multiple cyclins D, E, A and B 6) D and E cyclins and their function, the Rb and E2F proteins (Figs. 20-33) * D type cyclins come from proto-oncogenes * Cyclin E is the principle imposter getting the cell past the restriction point * Cyclin D-Cdk4 or 6 is activated first and then Cyclin E-Cdk2 * Once CyclinD-Cdk4/6 is activated in phosphorylates retinoblastoma protein (Rb) which releases E2F * E2F now acts as a transcription activator. Cyclin E-Cdk2 then phosphorylated even more Rb/E2F via positive feedback loop 17) Overview of mammalian check points, p53 (Figs. 20-34 and 20-35)