Practice Exam III Week 9 (04/08-04/09) Take a deep breath. We know you can do it.
Be patient and read every question carefully before you answer any question. Answer what we ask you to answer. This practice exam is not entirely comprehensive. So remember to go back and review your earlier worksheets. Part I: True/False 1^)__F__ In mammalian cells the import of proteins into the mitochondria begins before the polypeptide chain is completely synthesized - in other words it occurs co-translationally. 2^)_T___ To function effectively, each transport vesicle must selectively take up only the appropriate proteins and must fuse only with the correct target membrane. 3^)_F___ After a protein enters the nucleus, its signal sequence is removed by the activity of a signal peptidase, which ensure that it will remain in the nucleus. 4^)__F__ Complementary Rab proteins on transport vesicles and target membranes bind to one another to allow transport vesicles to dock selectively at their appropriate target membranes. 5^)__T__ Diacylglycerol is an example of a second messenger used in cell signaling. 6^)___F_ The receptors involved in paracrine and synaptic signaling have the highest affinity for their respective receptors. 7^)_T___ The activity of any protein regulated by phosphorylation depends on the balance at any instant between the activities of the kinases that phosphorylate it and the phosphatases that dephosphorylate it. 8)^_F___ Binding of extracellular ligands to receptor tyrosine kinases (RTKs) activates the intracellular catalytic domain by propagating a conformational change across the lipid bilayer through a single transmembrane region. 9) T The TOM complex is required for the import of all nucleus-encoded mitochondrial proteins. The fate of a protein with no sorting signal is to remain in the cytosol.
Part II: Multiple Choice 1^) If you were to remove the ER retention signal from protein disulfide isomerase (PDI),, which is normally a soluble resident of the ER lumen, where would you expect the modified PDI to be found? a. the ER b. the Cytosol c. extracellular fluid d. a lysosome
They provide a mechanical stability to animal cells
. a G-protein with a delayed activation response d. a G-protein that was constantly inactivated c. ubiquitin 3^) The sorting of proteins to mitochondria and chloroplasts is: a.
8!) Actin filaments and microtubules share all of the following properties EXCEPT: a. the ER b. the nucleus c. IP3 is produced directly by cleavage of an inositol phospholipid without the incorporation of an additional phosphate group b. The most likely final location for this protein is: a. structured to have cytoplasmic domains that are enzymatically active 7) Which of the following is FALSE a.2^) Proteins that do not fold properly in the ER lumen are degraded in the cytosol by: a. They are assembled from subunits that are heterodimers e. able to phosphorylate G-proteins d. They are intrinsically polar structures c. co-translational b.can activate other molecules downstream of GPCRS. proteosomes c. dimerize to bring the kinase domains of their receptor tails together c. They can be cross-linked into bundles 9!) Which of the following is NOT a characteristic of intermediate filaments a. secreted 5^) Which of the following would be expected to happen in a mutated G protein that results in a decreased rate at which the signal causes the receptor to release GDP. signal peptidases d. The extracellular signal molecule acetylcholine has different effects on different cell types in an animal and often binds to different cell surface molecules on different cell types c. They are involved in cell motility b. are able to function as monomers b. They form the nuclear lamina b. but no affect on its affinity thereafter for GTP? a. d. gated transport 4^) Imagine a protein engineered with an ER signal sequence at its N-terminus and a nuclear localization signal near the C-terminus. Calmodulin modulates the intracellular Ca2+ concentration. the cytosol d. ribosomes b. Both the GTP bound alpha subunits and nucleotide free beta gamma complexes--but not GDP bound fully assembled G-proteins -. a G-protein with a delayed inactivation response 6) Receptor tyrosine kinases a. post-translational c. They can associate with motor proteins d. pre-translational d. a G-protein that was constantly activated b.
However. d. The complexity of the microtubule plus end b. is required to initiate cytokinesis d. is driven by an energetically favorable process %13.
. c. d. It requires motor-driven microtubule sliding. alternate between high affinity and low affinity for microtubules. Both are dependent on rates of polymerization and depolymerization of subunits. The difference in critical concentration of GTP -tubulin and GDP-tubulin at the microtubule plus end c. Soon all the receptors full of cargo will be in the nucleus. Proteins will be imported into the nucleus normally. More than one of the answers above is correct %11.) Myosin motors require ATP hydrolysis to move along filaments because ATP hydrolysis a. Both are influenced by the concentration of subunits in the surrounding environment.) Which of the following statement(s) are FALSE? a. is important to promote actin polymerization b. It requires polymerization of intermediate filament fibers. Actin can generate force without the aid of motors. In one case the filament changes in size dramatically while in the other the size appears to remain the same. the receptor will not release the cargo. Imagine a mutant cell in which the Ran protein involved in nuclear transport can no longer bind to GTP. Some actin binding proteins can promote the formation of branched actin filaments. It requires polymerization and depolymerization of microtubules. b.) Two properties of microtubule polymerization are dynamic instability and treadmilling. Long actin filaments work best for pushing the plasma membrane during cell crawling.) Dyneins and kinesins a. Actin filaments underlie the plasma membrane in animal cells. w/o RAN-GTP in the nucleus. Which of the following is NOT a true statement about these processes? a.) Which of the following is NOT true of chromosome segregation during mitosis? a. is required to generate the force necessary for the power stroke c. Both can be influenced by various classes of actin-binding proteins. They include the keratin filaments of epithelial cells 10!) Dynamic instability of microtubules stems from a. if at all. by this mutation. %14.c.and minus end. b. Part III: Protein Localization ^1. They are composed of globular monomers that polymerize to form fibers e. have sequence and structural homology b. always transport cargo within the cell. %12. %15. b. The intrinsic ability of the tubulin subunit to hydrolyze GTP to GDP e. c. d. c. leaving no receptors to transport new proteins into the nucleus from the cytosol. d. are important during cytokinesis c. What is the phenotype or expected outcome for this mutant cell? Please be specific but be brief! Explain what part of nuclear transport will be affected. Their protein composition is tissue specific d. The fact that one of the GTPs on the tubulin dimer is never hydrolyzed d.directed molecular motors. It uses plus end.
You find that cells that lack the GPCR are 15% smaller than normal cells. or no change) if cells were treated in the following fashion. smaller. whereas cells that express a mutant. The G protein downstream of this receptor activates adenylyl cyclase. explain what you predict would happen to the cell’s size (bigger. d. c. which ultimately leads to the activation of PKA. A signal for import into the nucleus and a signal for the import into mitochondria. Part IV: Cell Signaling 1. Furthermore. which has been shown to inhibit certain subclasses of the a subunit of the G protein. A signal for import into the nucleus and a signal for import into the ER. A signal for import into mitochondria and a signal for retention in the ER.
. b. Given these results. If the genes were expressed in a cell. predict which signal would win out for each of the following combinations (please circle one signal). a.#2. the normal blood cells become smaller when treated with cholera toxin. A signal for import into the nucleus and a signal for export from the nucleus. each encoding a protein with a pair of conflicting signal sequences specifying different compartments.Briefly describe what they are and where in the GPCR signaling pathway the following components come into play: DAG
AC (adenylyl cyclase)
2) You are interested in cell size regulation and discover that signaling through a GPCR is important in controlling cell size in rat white blood cells. constitutively activated version of PKA are 15% larger than normal cells. Imagine that you engineered a set of genes.
so that the PKA binds more tightly to cAMP. Bigger. dynein and kinesin are motor proteins that use the energy of to move unidirectionally along microtubules.a.
e. You do not know the steps in the middle.You add a drug that inhibits adenylyl cyclase. A concentrated network of actin filaments underneath the plasma membrane forms the and is responsible for cell shape and movement. You add a drug that increases the activity of cyclic AMP phosphodiesterase. It would inhibit subclasses of the subunit of the G Protein. This is a form of chemotaxis.You mutate the cAMP-binding sites in the regulatory subunits of PKA. No change. motor proteins use as energy to move along actin filaments. You inhibit the RGS protein that normally works on the a subunit of the G protein involved in this pathway.
c. or cell movement in a direction controlled by a chemical gradient. The direction of movement is determined by the concentration of the microtubules. Smaller. You add pertussis toxin. Smaller.
. (Hint. Fill in the blanks: A.
3) Outline a general signaling pathway that would allow a yeast to move in response to yeast mating factor. Microtubules in cilia and flagella have a unique _________ configuration and interact with a special motor protein called . B. Eukaryotic cilia and flagella contain bundles of stable intermediate filaments that originate from structures called .
d. No change. causing adjacent filaments to slide past each other in structure known as . but you can predict generally how such a pathway would begin and end. how would the cell receive the mating factor signal and how would the cell be able to move towards it?)
Part V: Cytoskeleton & Motor Proteins !1.
% of actin subunits in filaments
Time A.!2. Steady State (Equilibrium)
C. Indicate on the figure what the graph would look like if a large amount of a nucleating protein was added at the start? Label this curve “C”. II and III. But dynein moves things toward the . I. The figure below represents a typical time course of polymerization of filamentous actin from actin monomers. About kinesin and dynein. D. B. 2) naming each of the three phases represented by numerals I. Explain (in 1-2 sentences) the behavior of monomers in each of the three phases above. Elongation (Growth Phase)
III.ends of MT's and kinesin moves things toward the + ends.
%3. a protein that sequestered (bound up) actin monomers and prevented polymerization? Assume enough of this protein is added to bind up half of /
. Nucleation (Lag Phase)
II. Complete the figure by 1) labeling the axes. Does dynein move things towards the nucleus of a cell and therefore towards a cell if it is coming from the outside? And does kinesin move things to the plasma membrane of a cell and therefore away? Both only move things inside cells. What would happen to the curve if you added nibblerin.
taken from Cell Bio PLUMS. 2011
. however. Cell BioExam III.%4. At that point. stabilizing the polymer). !. you set up multiple microtubule motility assays to get a truly accurate measure of kinetic parameters for kinesin. you adhered kinesin to a glass slide and added fluorescent microtubules so that they could glide along. propelled by the adhered kinesins. kinesin can carry vesicles for long distances along microtubule tracks. The first time she does. 2008. How does lateral association promote the subsequent formation of a microtubule?
%. wanting to encourage your quest for knowledge and willingness to “do what no one else will do”. Do you think the two head domains of a kinesin molecule are essential to accomplish this task or could a one headed kinesin function just as well? Explain your answer.taken from Omar Quintero.
B. Explain briefly why you will be unable to obtain reliable measurements of microtubule velocity without taxol.
!5. To do this. In a cell. The critical event for tubule formation is thought to be the first lateral association. A. President Pasquerella. ^. the probabilities that the next dimer will bind laterally or to the end of the protofilament are about the same.taken from Sharon Stranford. she forgets to add taxol (a drug that binds tightly to microtubule. offers to help you. A solution of alpha beta tubulin dimer (free of other proteins) is thought to nucleate microtubules by forming a linear protofilament about 7 dimers in length. In your spare time while preparing for your exam. BIO 220 Exam 3. 2010.