mcb130-fa08-mt1-Hariharan-soln

mcb130-fa08-mt1-Hariharan-soln - l}: ' pmswea Name: SID:...

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Unformatted text preview: l}: ' pmswea Name: SID: Question 1 (18 points) — Organization of the cell (4 points) Name two organelles in animal cells that are enclosed by two membranes. qud w; ’ \Mi law CMovwlviq (6 points) Cellular dimensions: In each case, circle the number that is closest to: a) The size of a mamm ' red blood cell: 0.7um 70 um b) The ' ess of a lipid bilayer: SOnm c) The size of the select‘ ' filter of the K+ channel: 0.1nm 61% 10mm 500nm (8 points) Are the following statements ‘True’ or ‘False’ a) All ribosome subunit assembly occurs in the nucleolus FREQ (wok ’NME g“ wi’eotkovavafl vibehmm b) Cells that make mostly secreted proteins have a lot of smooth endoplasmic reticulum Pause c) The mitochondria in bacterial cells have their own DNA ;m\5& d) The nucleolus is separated from the remainder of the nucleus by a membrane Pods/L 9 Mum/a \LE‘1 Name: SID: Question 2 (18 points) Nucleocytoplasmic transport, methods in cell biology: A drug that specifically blocks the interaction of exportin with all cargoes is added to tissue culture cells. Shortly after addition of the drug, what would the effects be on: a) (2 points) The localization of newly assembled ribosomal subunits kceLmekk in MLVLuS b) (2 points) Export of mRNAs from the nucleus No «Max 0) (2 points) The export of importin from the nucleus to the cytoplasm No eldulr (6 points) Since there is mixing of nuclear and cytoplasmic proteins during mitosis, how is the nucleocytoplasmic gradient of Ran-GTP re-established when the nuclear envelope reassembles after mitosis? LAM/EQ? “south/W) WW wwwhw /d~ww+\\,, AWWB MIR“ (:1 Wm) (Numb) KM’Q‘W {‘3 MA" 6;;th cJ~Mu-~y~w<J (JAN '5 WM \(LJL Mel‘th welt/La??— xvn/MJ wow—A WMhMJ . . . . . '9— Huh) (6 pomts) In what way can we study trafficking in cells w1th green fluorescence protein (GFP) that we could not do previously with electron microscopy? In what way has it changed our View of cell structure? Gd"? 0 (wk-«SKULL: vaNth «All LM LA fig, SWEflwJL> (Lt— MNAD LAX-Aw Vlwul'wd ML \Mva oLJKaMLAvLM V, X- Name: SID: Question 3 (18 points) Membrane proteins a) (9 points) Use the membrane diagram below to draw how the mature LDL receptor, cytochrome P450 and BiP are arranged within the ER membrane and the ER lumen. Indicate the amino- and carboxy-terminus of each protein. '60 fl 0‘ 0Q“ 03° 06} Qoq \<2 \s x« & °° cQ 0“ '9“ :9 «5‘ s (a 9 I 20 500 520 900 I 100 120 400 I 20 700 [Note: numbers indicate amino acid positions] \0 ov‘ ( vaerQM} (PM) \.9 {w WkMWWM \u'oo CYTOSOL ER MEMBRANE 3.0 W! ER LUMEN 700 H CMLK‘ \0 \5 me @ day/tug «gr $5390.52 HkAQ 2.0 ,f \ gawk mm? \ouwww ) \Dfivd' £34 Lew-Ct} MWM”) \ WW ‘64 C‘WWSL “K i-L Rfik'e‘kss who Amwflk La Name: SID: b) (9 points) Protein translocation of LDL receptor, cytochrome P450 and BiP is studied in an in vitro translocation assay. The assay takes advantage of (1) in vitro translation reactions (IVT) in the presence of rough microsomes (RM), or (2) in vitro translation reactions (IVT) in the presence of purified SRP (no rough microsomes). Assume that all factors necessary for translation are present in both cases. In each case, the proteins obtained are separated on a gel (SDS-PAGE). Use the gel below to indicate the position and label the molecular weight of each of the translation products that you expect in each reaction. For simplicity, assume that each amino acid has a molecular weight of 100 Da. IVT+ RM |VT+ SRP 45,? 69 "0‘ “0‘ 3.9 3: K ET “4 ° W o ‘5 _| - 9: .1 :2 Isl [6‘] In [El [5‘ 100 '— 90 _ my.» 88 — ’ 86 —- 80 - 70 _ SWIM 68 — - 66 — 6O -— 50 — ’ How-bot 4o— 38— 35— 30-— wL whawmfilc 12— LC-bat \‘s actuatde 10-— MquK \Lfi Name: SID: Question 4: (18 points) Vesicle trafficking Are the following statements ‘True’ or ‘False’? a)(2 points) Transmembrane proteins that are resident in the ER have a KDEL motif gods L b) (2 points) Dynamin is required for COPII-coated vesicles to bud off the ER pmlJL c) (2 points) Patients with I—cell disease secrete all of their lysosomal enzymes into the extracellular fluid TV“ L d) (2 points) Individual clathrin—coated vesicles that form at the plasma membrane can simultaneously contain both LDL-receptor and transferrin receptor molecules Time. e) (2 points) The binding of v-snares and t-snares is facilitated by NSF and ATP hydrolysis Fa,\§L—— f) (4 points) Compare what happens to the LDL-receptor and its ligand with what happens to the transferrin-receptor and its ligand in the low pH of the endosome. we ouswv‘aia «(M m LDL uan M rub Wyn“ - Tmsxbwi‘x vawm‘vx} W vcuafw M dew) M fie L,th lo W», g) (6 points) Explain the cistemal maturation model of trafficking cargoes through the Golgi apparatus i.e. in this model, what happens to cargoes and what happens to resident Golgi enzymes? 3WDQA-l‘xh (L {by CCSXOVH— (MW M W WM wa ugeMLaA‘.& alt/m; W)? OLA/8&1} QM Hm stWon-L 0.) “*4? WM l‘o Ew Mflrkdrowa deka \A Lad/u C's/975m. M < ‘ >0‘VL Rm)owM Vzwcxe) r\-o uskwu M QM \yz/HA — 3N» Owe—7p Sm WWW» 5&6“; Lute—«M- MNQK \LC/j Name: SID: M V" Question 5: (1% points) Putting it all together. . .. You perform a FLIP (fluorescence loss in photobleaching) experiment on a cell where the low-density lipoprotein receptor (LDL receptor) is fluorescently labeled. You bleach and measure from different regions of the cell membrane. Fluorescence BLEACH Bleaching area Measurement area a) (2 points) On the axes provided, draw the result you would expect for a FLIP experiment conducted at 37°C. Label this line ‘37°C’. b)(2 points) On the same axes, draw the result you would expect for a FLIP experiment conducted at 4°C. Label this line ‘4°C’. c)(6 points) On the same axes, draw the result you would expect if the experiment was done at 37°C in the presence of a drug that inhibits dynamin function. Label this v line ‘+ dynamin’ Explain why this line is or is not the same as the lines drawn in 63%: ) 0:22,: uth \,\oc}c M We WW D& vs U—filcwfi gym“ M‘k Vflfik‘efi 3:0 Md?— oM «AK sadlo c756: 9-1; M Vbjcfivj \yuk My AflR/SVKBOA (Aura; “\9 £51 Aquila-whoa 2— ?6wl‘) and it «3.) d) (8 points) What happens to LDL receptor levels in cells if the cells had been treated with a statin? (increase, decrease or unchanged). Explain the mechanism by which statins would have this effect. (a u SW» —«> \«mer ANG'CDPX wka .t and i > ‘MVMAL‘NN QMMS (m lowsz Low MW .t Noumea? 5) pr‘fihkvlm* 50k?"$£2§9 year» t 5“ 4‘48 5 UV: “4.. :KVVLMMM ‘ ' ‘ sr se%€ VWRK) VA W)\.-Ln.)orv\ 63’ {m as, vs. 9w mm» WWW; m Wt. what“ Ext—«9r ’5 ’l LDL ww—Xw Lewd), 36m $0 §._Ee§> H \ mam. % SW??? . W 89.x :95, SE35 3332.: mama—m Emnlcczo: S t n «8 u t S AND No-Nm mm-wo wTwm wmio Aoim Am-mo mo-mm mm-mo mo-mm mmlwo Vohm um-mo mo-mm mmbo moo—d ...
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This note was uploaded on 09/14/2011 for the course MCB 58180 taught by Professor Kuriyan during the Spring '11 term at University of California, Berkeley.

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mcb130-fa08-mt1-Hariharan-soln - l}: ' pmswea Name: SID:...

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