14 Intracellular Compartments

and Transport
Questions
MEMBRANE-BOUNDED ORGANELLES (Pages 448–452)
Eucaryotic Cells Contain a Basic Set of Membrane-bounded Organelles (Pages 449–450)

14–1 Easy, short answer

Name the membrane-bounded compartments in a eucaryotic cell where each of the functions
listed below takes place.
A. Photosynthesis.
B. Transcription.
C. Oxidative phosphorylation.
D. Modification of secreted proteins.
E. Steroid hormone synthesis.
F. Degradation of worn-out organelles.
G. New membrane synthesis.
H. Breakdown of lipids and toxic molecules.

14–2 Easy, multiple choice

Which organelles are most numerous in a typical animal cell such as a liver cell?
A. Peroxisomes.
B. Lysosomes.
C. Endosomes.
D. Mitochondria.
E. Golgi apparatus.

14–3 Easy, multiple choice

Which membrane-bounded compartment occupies the greatest volume (in total) in a typical
animal cell such as a liver cell?
A. Nucleus.
B. Cytosol.
C. Mitochondria.
D. Lysosomes.
E. Endoplasmic reticulum.

187
188 Essential Cell Biology Test Bank

Membrane-bounded Organelles Evolved in Different Ways (Pages 450–452)

14–4 Easy, short answer

You discover a fungus that contains a strange star-shaped organelle not found in any other
eucaryotic cell you have seen. On further investigation you find the following:
A. the organelle possesses a small genome in its interior.
B. the organelle is surrounded by two membranes.
C. vesicles do not pinch off the organelle membrane.
D. the interior of the organelle contains proteins similar to those of many
bacteria.
E. the interior of the organelle contains ribosomes.

How might this organelle have arisen?

14–5 Easy, art labeling

Complete diagrams B and C in (A) (B) (C)
Figure Q14–5 to show how the
nucleus and endoplasmic
reticulum might have evolved in DNA

an ancient procaryotic ancestor

of the eucaryotic cell. (You do not
need to sketch any of the other
organelles in the cell.) Label the
different features (membranes,
compartments) in the ancient
eucaryotic cell. ancient ancient
procaryotic cell eucaryotic cell
Q14–5

PROTEIN SORTING (Pages 452–462)

Proteins Are Imported into Organelles by Three Mechanisms (Page 453)

14–6 Easy, matching/fill in blanks

For each of the following sentences, fill in the blanks with the correct word or phrase selected
from the list below. Use each word or phrase only once.
A. Plasma membrane proteins are inserted into the membrane in the ________.
B. The address information for protein sorting in a eucaryotic cell is contained in
the ________ of the proteins.
C. Proteins enter the nucleus in their ________ form.
D. Proteins that remain in the cytosol do not contain a ________.
E. Proteins are transported into the Golgi apparatus via ________.
F. The proteins transported into the endoplasmic reticulum by ________ are in
their ________ form.

protein translocators; amino acid sequence; endoplasmic reticulum; plasma

membrane; sorting signal; folded; unfolded; transport vesicles; Golgi apparatus.
Chapter 14: Intracellular Compartments and Transport 189

Signal Sequences Direct Proteins to the Correct Compartment (Pages 453–455)

14–7 Intermediate, short answer (Requires information from section on pages

458–459)
What would happen in each of the following cases? Assume in each case that the protein
involved is a soluble protein, not a membrane protein.
(A) You add a signal sequence (for the ER) to the amino-terminal end of a
normally cytosolic protein.
(B) You change the hydrophobic amino acids in an ER signal sequence into
charged amino acids.
(C) You change the hydrophobic amino acids in an ER signal sequence into other,
hydrophobic, amino acids.
(D) You move the amino-terminal ER signal sequence to the carboxyl-terminal end
of the protein.

14–8 Intermediate/difficult, short answer

You are trying to identify the peroxisome- no histidine
targeting sequence in the peroxisomal +
enzyme thiolase from yeast. To this end, 1 200
thiolase –
you have created a variety of hybrid genes
50 200
that encode hybrid proteins containing part –
of the thiolase attached to another protein,
100 200
histidinol dehydrogenase (HDH), which is a –
cytosolic enzyme required for the synthesis 125 200
of the amino acid histidine. You genetically –
engineer a series of yeast cells to express 150 200
+
these hybrid proteins instead of their own
1 150
versions of the enzymes. If the hybrid pro- –
tein is imported into the peroxisome, the 1 100
HDH portion cannot function properly and –
the yeast is unable to grow on medium lack- 1 50
ing histidine. You obtain the results shown –
in Figure Q14–8. Where are the peroxisomal 1 25
+
targeting sequence(s) in thiolase?
The numbers above each protein indicate the amino
acids from thiolase present in the hybrid protein.
+ indicates growth on medium lacking histidine;
Q14–8 – indicates lack of growth on medium lacking histidine.

Proteins Enter the Nucleus Through Nuclear Pores (Pages 455–457)

14–9 Easy, multiple choice

Which of the following are routinely both imported into and exported from the nucleus?
A. Histones.
B. Gene regulatory proteins.
C. DNA polymerases.
D. Ribosomal proteins.
E. Nuclear lamina proteins.
190 Essential Cell Biology Test Bank

14–10 Intermediate, multiple choice

The cytoplasms of adjoining plant cells are connected by fine channels called plasmodesmata
whose structure is shown in Figure Q14–10:
Movement of proteins through plasmodesmata is likely to be most similar to movement of
proteins:
A. from the cytoplasm into a mitochondrion.
B. from the cytoplasm into the ER.
C. from the ER to the Golgi aparatus.
D. from the nucleus into the cytoplasm.
E. from the cytoplasm into a peroxisome.

cell 1 cell 2

nucleus nucleus

plasmodesmata
pore proteins
cell wall
plasma
plasmodesmata membrane
cytoplasm
Q14–10

14–11 Easy, multiple choice

What is the role of the nuclear localization sequence in a nuclear protein?
A. It is bound by cytoplasmic proteins that direct the nuclear protein to the
nuclear pore.
B. It is a hydrophobic sequence that enables the protein to enter the nuclear
membranes.
C. It aids protein unfolding in order for the protein to thread through nuclear
pores.
D. It prevents the protein diffusing out of the nucleus via nuclear pores.
E. It directs the protein to the nuclear lamina.

14–12 Difficult, data interpretation (Requires information from Chapter 5 panels)

A gene regulatory protein, A, contains a typical nuclear localization signal but suprisingly is
usually found in the cytosol of a cell. When the cell is exposed to hormones, however, protein A
moves from the cytoplasm into the nucleus where it activates genes involved in cell prolifera-
tion. When you purify protein A from unstimulated cells, you find that another protein, protein
B, is complexed with it. To determine the function of protein B, you make mutants lacking the
gene for protein B. You then fractionate extracts of gently lysed normal and B mutant cells into
cytoplasmic and nuclear fractions, separate the proteins in these fractions by gel electrophore-
sis, and probe the gels for the presence of proteins A and B using Western blotting techniques.
Your results are shown in Figure Q14–12.
Chapter 14: Intracellular Compartments and Transport 191

On the basis of these results, which is the most likely function of Protein B? Explain your rea-
soning.
A. In the presence of hormone, Protein B neutralizes the charges on Protein A’s
nuclear localization signal by covalently attaching hydrophobic groups to the
lysine side chains.
B. In the absence of hormone, protein B cleaves the nuclear localization signal off
protein A.
C. In the absence of hormone, Protein B binds to the nuclear localization signal
on protein A, blocking its action.
D. Protein B is a nuclear import receptor.
E. Protein B prevents Protein A from unfolding.

Normal Lacking protein B

Unstimulated Stimulated Unstimulated Stimulated

cytopl. Nucl. cytopl. Nucl. cytopl. Nucl. cytopl. Nucl.

Protein A

Protein B
Q14–12

Proteins Unfold to Enter Mitochondria and Chloroplasts (Pages 457–458)

14–13 Easy, multiple choice

Which of the following statements about import of proteins into mitochondria are true?
A. The signal sequences on mitochondrial proteins are usually carboxyl terminal.
B. The first stage of import of a mitochondrial protein is across the outer mem-
brane into the intermembrane space.
C. Most mitochondrial proteins are not imported from the cytosol but are synthe-
sized inside the mitochondria.
D. Mitochondrial proteins are translocated across the inner and outer mem-
branes simultaneously.
E. Mitochondrial proteins cross the membrane in an unfolded state.

14–14 Intermediate/difficult, multiple choice (Requires information from

Chapter 5)
Which of the following will inhibit import of an enzyme into mitochondria?
A. Chaperone proteins.
B. A small molecule that binds tightly to the active site of the enzyme.
C. Inhibition of ATP synthesis.
D. A high concentration of free mitochondrial signal peptide in the cytosol.
E. An inhibitor of mitochondrial signal peptidase.
192 Essential Cell Biology Test Bank

Proteins Enter the Endoplasmic Reticulum While Being Synthesized (Pages 458–459)

14–15 Easy, multiple choice

Proteins destined to enter the endoplasmic reticulum:
A. are transported across the membrane after their synthesis is complete.
B. are synthesized on free ribosomes in the cytosol.
C. begin to cross the membrane while still being synthesized.
D. cross the membrane in a folded state.
E. all remain within the endoplasmic reticulum.

14–16 Easy, multiple choice

After isolating the rough endoplasmic reticulum from the rest of the cytoplasm, you purify the
RNAs attached to it. What proteins do you expect these RNAs to encode?
A. Soluble secreted proteins.
B. ER membrane proteins.
C. Mitochondrial membrane proteins.
D. Plasma membrane proteins.
E. Ribosomal proteins.

Soluble Proteins Are Released into the ER Lumen (Pages 459–460)

14–17 Intermediate, multiple choice + short answer

You are studying the in vitro translation and import of proteins into isolated vesicles made
from the ER (microsomes). Using differential centrifugation you have separated the cytoplasm
of the cell type from which the ER originally came into several different fractions. One of these
fractions stimulates the import of proteins into microsomes if the microsomes are added after
translation is completed. It has no effect on protein import when translation takes place in the
presence of the microsomes. Which of the following are most likely to be present in this frac-
tion? Explain your answer.
A. Nuclear import receptor.
B. SRP receptor.
C. Signal peptidase.
D. Chaperone protein.
E. Ribosomes.

Start and Stop Signals Determine the Arrangement of a Transmembrane Protein in the Lipid Bilayer
(Pages 461–462)

14–18 Easy, short answer

Briefly describe the mechanism by which the presence of an internal stop-transfer sequence in
a protein causes the protein to become embedded in the lipid bilayer as a transmembrane
protein with a single-membrane-spanning region? Assume that the protein has an amino ter-
minal signal sequence and just one internal hydrophobic stop-transfer sequence.
Chapter 14: Intracellular Compartments and Transport 193

14–19 Intermediate/difficult, art labeling

A plasma membrane protein X traverses the membrane three times in the orientation shown
in Figure Q14–19. (N = amino terminus; C = carboxyl terminus.) The hydrophobic membrane-
spanning regions are shown as open boxes.
(A) Sketch the arrangement of the newly synthesized protein chain after it has
completed its entry into the ER membrane but before any action of signal pep-
tidase. Label the cytosol and ER lumen in your diagram.
(B) Label the location of stop-transfer (S) and start-transfer (T) sequences on your
diagram.
N

extracellular
space

cytosol

Q14–19

14–20 Intermediate/difficult, multiple choice (Note to instructors: this is an alter-

native question to 14–19 that does not require the student to sketch any-
thing; giving them both in the same test will give away the answer to 14–19)
Figure Q14–20B shows the orientation of a multipass membrane protein in the plasma mem-
brane. This protein also had an amino-terminal signal sequence, which was cleaved off in the
endoplasmic reticulum (Figure Q14–20A). All of the membrane-spanning regions (depicted as
open boxes) in this protein have the same amino acid sequence, leading to the hypothesis that
any hydrophobic signal sequence has the potential to act as either a start or a stop signal; the
actual behavior of the signal is determined by the position of the signal relative to other signals
in the protein. If this hypothesis were true, which of the following modifications would cause
the protein depicted in Figure Q14–20B to be inserted in the membrane in exactly the opposite
orientation relative to the cytoplasm?
A. Changing hydrophobic amino acids in the first, cleaved, sequence to charged
amino acids.
B. Changing hydrophobic residues in every other signal sequence to charged
residues, starting with the first, cleaved, signal sequence.
C. Adding a new signal sequence and signal peptidase recognition site to the car-
boxyl terminus of the protein.
D. Deleting the first signal sequence.

signal
peptidase N

N
C C

Q14–20 (A) (B)

194 Essential Cell Biology Test Bank

VESICULAR TRANSPORT (Pages 462–467)

Transport Vesicles Carry Soluble Proteins and Membrane Between Compartments (Page 463)

14–21 Easy, matching/fill in blanks

For each of the following sentences, fill in the blanks with the correct word or phrase selected
from the list below. Use each word or phrase only once.
A. Proteins are transported out of a cell via the ________ or ________ pathway.
B. Fluid and macromolecular material is transported into the cell via the ________
pathway.
C. All proteins being transported out of the cell pass through ________ and
________.
D. Transport vesicles link organelles of the ________ system.
E. The formation of ________ in the endoplasmic reticulum stabilizes protein
structure.

the endoplasmic reticulum; endomembrane; carbohydrate; secretory; endocytic; the Golgi

apparatus; endosome; lysosome; protein; disulfide bonds; exocytic; hydrogen bonds; ionic
bonds.

14–22 Easy, multiple choice

An individual transport vesicle:
A. contains only one type of protein in its lumen.
B. will fuse with only one type of membrane.
C. is endocytic if it is traveling toward the plasma membrane.
D. is enclosed by a membrane with the same lipid and protein composition as the
membrane of the donor organelle.

Vesicle Budding Is Driven by the Assembly of a Protein Coat (Pages 463–465)

14–23 Easy, multiple choice

Clathrin-coated vesicles do NOT:
A. bud from the plasma membrane.
B. bud from the endoplasmic reticulum.
C. bud from the Golgi apparatus.
D. require GTP hydrolysis for their budding.
E. contain adaptin coat proteins.

14–24 Easy, multiple choice

Adaptin proteins:
A. are found in both clathrin-coated and COP vesicles.
B. can hydrolyze GTP.
Chapter 14: Intracellular Compartments and Transport 195

C. form a basketlike network on the cytosolic side of the membrane.

D. remain on the vesicle after clathrin coat disassembly.
E. bind to different types of cargo receptors.

14–25 Intermediate, short answer

Clathrin-coated vesicles will bud from plasma membrane fragments when adaptins, clathrin,
and dynamin-GTP are added. What would you observe in each case if you omitted:
(A) dynamin?
(B) adaptins?
(C) clathrin?

The Specificity of Vesicle Docking Depends on SNAREs (Pages 465–467)

14–26 Easy, multiple choice

vSNARES participate directly in:
A. assembly or formation of the transport vesicles.
B. movement of the vesicle along cytoskeletal filaments.
C. uncoating of the vesicle.
D. docking of the vesicle to the target organelle.
E. fusion of the vesicle to the target organelle.

SECRETORY PATHWAYS (Pages 467–472)

Most Proteins Are Covalently Modified in the ER (Pages 467–468)

14–27 Easy, multiple choice

N -linked oligosaccharides on secreted glycoproteins are attached to:
A. nitrogen atoms in the polypeptide backbone.
B. the serine or threonine in the sequence Asn-X-Ser/Thr.
C. the amino terminus of the protein.
D. the asparagine in the sequence Asn-X-Ser/Thr.
E. the aspartic acid in the sequence Asp-X-Ser/Thr.

14–28 Easy, multiple choice

Which of the following statements about glycosylation are true?
A. Cytosolic proteins are never glycosylated.
B. Sugars are transferred one by one from dolichol to protein.
C. The oligosaccharide transferred from dolichol is composed only of mannose.
196 Essential Cell Biology Test Bank

D. The oligosaccharide on a protein leaving the endoplasmic reticulum usually

differs from the oligosaccharide transferred to that protein from dolichol.
E. Glycosylation occurs only after the protein has completed its entry into the
endoplasmic reticulum.

Proteins Are Further Modified and Sorted in the Golgi Apparatus (Pages 469–470)

14–29 Easy, art labeling

Match the set of labels below with the
numbered label lines on Figure Q14–29. 3

A. Cisterna.
2
B. Golgi stack.
plasma
C. Secretory vesicle. membrane

D. trans Golgi network.

E. cis Golgi network.
4
5

1
Q14–29

14–30 Intermediate/difficult, data interpretation

A plasma membrane protein carries an oligosaccharide containing mannose (Man), galactose
(Gal), sialic acid (SA), and N- acetylglucosamine (GlcNAc). These sugars are added to the pro-
tein as it proceeds through the secretory pathway. First, a core oligosaccharide containing Man
and GlcNAc is added, followed by Gal, Man, SA, and GlcNAc in a particular order. Each addi-
tion is catalyzed by a different transferase acting at a different stage as the protein proceeds
through the secretory pathway. You have isolated mutants defective for each of the transferas-
es, purified the membrane protein from each of the mutants, and identified which sugars are
present in each mutant protein. The results are summarized in Table Q14–30.

Table Q14–30
Sugars present in the purified protein
Cell lacking: Man Gal SA GlcNAc
A. Oligosaccharide – – – –
protein transferase
B. Galactose + – – +
transferase
C. SA transferase + + – +
D. GlcNAc + – – less than in
transferase normal cells
Chapter 14: Intracellular Compartments and Transport 197

From these results, match each of the transferases (A, B, C, D) to its subcellular location select-
ed from the list below. (Assume that each location contains only one enzyme.)
1. Central Golgi cisternae.
2. cis Golgi network.
3. ER.
4. trans Golgi network.

14–31 Easy, matching/fill in blanks

Complete each of the following sentences correctly by crossing out one of the alternatives in
brackets.
A. New plasma membrane reaches the plasma membrane by the [regulated /
constitutive] exocytosis pathway.
B. New plasma membrane proteins reach the plasma membrane by the
[regulated / constitutive] exocytosis pathway.
C. Insulin is secreted from pancreatic cells by the [regulated / constitutive] exocy-
tosis pathway.
D. The interior of the trans Golgi network is [acidic / alkaline].
E. Proteins that are constitutively secreted [aggregate / do not aggregate] in the
trans Golgi network.

Secretory Proteins Are Released from the Cell by Exocytosis (Pages 470–472)

14–32 Intermediate, multiple choice

You have purified insulin-containing vesicles from pancreatic cells and exocytic vesicles from
another cell type not specialized for secretion. In which of the following ways do you expect
the two types of vesicles to be similar?
A. The same tSNAREs mark the cytoplasmic face of the vesicle membranes.
B. They will have the same concentration of protein in their lumens.
C. They are likely to have a similar concentration of Ca2+ in their lumens.
D. The same cytoplasmic fractions must be added in order to stimulate fusion
with the plasma membrane.
E. They will be the same size.

ENDOCYTIC PATHWAYS (Pages 472–477)

Specialized Phagocytic Cells Ingest Large Particles (Pages 472–473)
Fluid and Macromolecules Are Taken Up by Pinocytosis (Pages 473–474)

14–33 Easy, multiple choice

Which of the following statements are true?
A. Pinocytosis and phagocytosis are the two major forms of exocytosis.
B. All eucaryotic cells ingest fluid by pinocytosis and do so at approximately the
same rate.
198 Essential Cell Biology Test Bank

C. Macrophages will endocytose large particles only if the particles bind to recep-
tors on the phagocytic cell surface.
D. Both phagocytic and pinocytic vesicles begin assembly as clathrin-coated pits.
E. Phagocytic vesicles fuse with the endosome; pinocytic vesicles with the Golgi
apparatus.

Receptor-mediated Endocytosis Provides a Specific Route into Animal Cells (Pages 474–475)

14–34 Difficult, data interpretation + multiple choice (Requires information from

section on pages 475–476)
You have always had trouble keeping your blood cholesterol levels within a healthy range,
whereas your sister has always been able to eat what she liked with impunity. To find out the
reason, you compare the ability of your cells to take up LDL compared to your sister’s cells. To
do this you add LDL particles from your sister’s blood to a sample of your sister’s cells and to a
sample of your own cells, and then wash off the LDL that has not become bound to the LDL
receptor. You then wait either 2 minutes or 40 minutes, and treat half the cells in each sample
with a protein-digesting enzyme (a protease). You then lyse the treated cells and determine the
presence or absence of LDL protein. The results are given in Table Q14–34.

Table Q14–34
After 2 minutes After 40 minutes
Sister You Sister You
Treated with protease + + – –
Not treated with protease + + – +

+ indicates LDL protein detected; – indicates LDL protein not detected

On the basis of these results, which of the following is the most likely explanation for your ten-
dency to a high blood cholesterol? Explain your answer.
A. Although you did not test binding of your own LDL particle, you can deduce
the only defect you could have must be a mutation in your own LDL protein
that prevents it from binding to the receptor.
B. Your cells have fewer LDL receptors than do your sister’s cells.
C. Your LDL receptors cannot bind to LDL.
D. Your LDL receptors bind to LDL but cannot be localized to clathrin-coated pits
and internalized.
E. Your LDL receptors cannot release LDL at low pH.

Endocytosed Macromolecules Are Sorted in Endosomes (Pages 475–476)

14–35 Easy, short answer

Name three possible fates for an endocytosed molecule that has reached the endosome.
Chapter 14: Intracellular Compartments and Transport 199

Lysosomes Are the Principal Sites of Intracellular Digestion (Pages 476–477)

14–36 Difficult, data interpretation

Fibroblast cells from patients W, X, Y and Z, who each have a different inherited
defect, all contain “inclusion bodies,” which are lysosomes filled with undi-
gested material. You wish to identify the cellular basis of these defects. The
possibilities are:
1. a defect in one of the lysosomal hydrolases.
2. a defect in the phosphotransferase that is required for mannose-6-phosphate
tagging.
3. a defect in the mannose-6-phosphate receptor, which binds lysosomal pro-
teins in the trans Golgi network and delivers them to lysosomes.
You find that when some of these mutant fibroblasts are incubated in media in which normal
cells have been grown, the inclusion bodies disappear. This leads you to suspect that lysoso-
mal hydrolases can be secreted by the constitutive exocytic pathway in normal cells and are
being taken up by the mutant cells. (It is known that some mannose-6-phosphate receptor
molecules are found in the plasma membrane and can take up and deliver lysosomal proteins
via the endocytic pathway.) You incubate cells from each patient with media from normal cells
and media from each of the other mutant cell cultures, and get the following results.

Media
From From From From From
normal cultures cultures cultures cultures
cells of W cells of X cells of Y cells of Z cells
Cell Line
Normal + + + + +
W – – – – –
X + + – – –
Y + + – – +
Z + + – + –

+ indicates that the cells appear normal; – indicates that the cells still have inclusion bodies.
For each patient (W, X, Y, Z) indicate which of the defects (1, 2, 3) they are most likely to have.
200 Essential Cell Biology Test Bank

Answers
A14–1. A. Photosynthesis = chloroplast.
B. Transcription = nucleus.
C. Oxidative phosphorylation = mitochondrion.
D. Modification of secreted proteins = Golgi apparatus and rough endoplasmic reticulum (ER).
E. Steroid hormone synthesis = smooth ER.
F. Degradation of worn-out organelles = lysosome.
G. New membrane synthesis = ER.
F. Breakdown of lipids and toxic molecules = peroxisome.

A14–2. D.

A14–3. B.

A14–4. A genome, a double membrane, ribosomes, and proteins similar to those found in bacteria are
evidence for an organelle having evolved from an engulfed bacterium.

A14–5. Figure A14–5. inner nuclear membrane

nuclear outer nuclear membrane
pore
nucleus

endoplasmic
reticulum membrane

endoplasmic
reticulum lumen

cytosol

ancient
eucaryotic cell
A14–5

A14–6. A. Plasma membrane proteins are inserted into the membrane in the endoplasmic reticulum.
B. The address information for protein sorting in a eucaryotic cell is contained in the amino
acid sequence of the proteins.
C. Proteins enter the nucleus in their folded form.
D. Proteins that remain in the cytosol do not contain a sorting signal.
E. Proteins are transported into the Golgi apparatus via transport vesicles.
F. The proteins transported into the endoplasmic reticulum by protein translocators are in an
unfolded form.

A14–7. (A) The protein will now be transported into the ER lumen. (B) The altered signal sequence will
not be recognized and the protein will remain in the cytosol. (C) The protein will still be deliv-
ered into the ER. It is the distribution of hydrophobic amino acids that is important, not the
actual sequence. (D) The protein will not enter the ER. Because the carboxyl terminus of the
protein is the last part to be made, the ribosomes synthesizing this protein will not be recog-
nized by the SRP and carried to the ER.

A14–8. From the results, the sequences 1–50 and 125–200 are the smallest pieces of thiolase that are
sufficient to cause HDH to be transported into the peroxisome; hence each of these pieces
Chapter 14: Intracellular Compartments and Transport 201

must contain a signal sequence. It is important to realize that even though 1–25 and 150–200
are not sufficient on their own to cause import into the peroxisome, we cannot tell from these
data whether these sequences are necessary for import as part of the larger sequence. In other
words, we cannot say that 25–50 and 125–150 contain complete signal sequences as a signal
sequence might be located, for example, from 20–30 or from 145–155. Thus it is not possible to
identify the target minimum sequence(s) from this experiment.

A14–9. D. Ribosomal proteins are imported from the cytoplasm, where they are synthesized, and
exported as part of ribosomal subunits, which are assembled in the nucleus.

A14–10. D. Transport across the plasmodesmata involves crossing two contiguous membranes through
a protein complex large enough to see in the electron microscope. This is most similar to
transport through nuclear pores.

A14–11. A.

A14–12. C. Protein B could work by preventing the nuclear localization signal from binding to nuclear
import receptors; this could be accomplished by either reversibly modifying the signal or by
binding to the signal and shielding it from the nuclear import receptor. Since protein A is sta-
bly associated with protein B in unstimulated cells, C is the most likely answer. In unstimulat-
ed cells, both protein A and protein B are associated with one another in the cytoplasm. In
stimulated cells, protein A but not protein B moves into the nucleus. In cells lacking protein B,
protein A is nuclear both in the presence and absence of hormone. This tells us that protein B
is required for keeping protein A out of the nucleus. Hence, it seems unlikely that protein B is a
nuclear import receptor. Protein B is unlikely to act by preventing protein A from unfolding
because nuclear import does not require unfolding. If protein B acted by cleaving the nuclear
localization signal off protein A, we would expect cytoplasmic protein A to be smaller than
nuclear protein A and thus show up as a band in a slightly different position in the gel. In addi-
tion, it would be extremely unlikely that protein A could regain a nuclear localization signal
upon hormone stimulation.

A14–13. D and E.

A14–14. B, C, and D. Proteins must unfold in order to cross the mitochondrial membrane. Since the
enzyme active site is a property of the folded protein, a small molecule that binds tightly to the
active site (B) will stabilize the folded protein, prevent unfolding and inhibit import.
Mitochondrial import is an active process and therefore requires ATP (C). A large concentra-
tion of the signal peptide would compete with the full-length enzyme for the receptor and
should therefore inhibit import (D). Chaperone proteins (A) promote both folding and unfold-
ing of the polypeptide chain and therefore would be expected to promote import of the
enzyme. Cleavage of the signal is not required for import, so inhibition of the signal peptidase
will not affect import (E).

A14–15. C.

A14–16. A, B, and D. The rough ER consists of ER membranes and polyribosomes that are in the
process of translating and translocating proteins into the ER membrane and lumen. Thus all
proteins that end up in the lysosome, Golgi apparatus, or plasma membrane, or are secreted,
will be encoded by the RNAs associated with the rough ER. Mitochondrial and ribosomal pro-
teins are translated on free cytosolic ribosomes.

A14–17. D. When the microsomes are added to proteins that have already been translated, the proteins
will have already folded and would need to unfold before they could be imported into the ER.
Hence, chaperones might be expected to facilitate import. They are not required when micro-
202 Essential Cell Biology Test Bank

somes are present during translation since the proteins that are being translated are immedi-
ately imported as they come off the ribosome before protein folding can take place. The pres-
ence of nuclear import receptors (A) is irrelevant for protein import into the ER. SRP receptor
(B) is a membrane protein and is present in the microsomes. Signal peptidase (C) is not
required for import into the lumen of the ER; it is required only for the final processing of the
protein. Ribosomes (E) will have no effect on proteins that have already been translated.

A14–18. The amino-terminal signal sequence initiates translocation and the protein chain starts to
thread through the translocation channel. When the stop-transfer sequence enters the translo-
cation channel, the channel discharges both the signal sequence and the stop-transfer
sequence sideways into the lipid bilayer. The signal sequence is then cleaved, so that the pro-
tein remains held in the membrane by the hydrophobic stop-transfer sequence.

A14–19. (A) Figure A14–19. Since the amino terminus (N) of the protein is on the extracellular side of
the plasma membrane, there must have been a signal sequence (acting as a start signal) at the
extreme amino terminus; this signal was cleaved off by signal peptidase in the ER lumen. (B)
Figure A14–19.
signal
peptidase

ER lumen

S T S
cytosol
N
C

A14–19

A14–20. D. Figure A14–20A. Deleting the first signal sequence completely would convert the original
first stop-transfer signal to an internal start-transfer signal and would both invert the protein
in the membrane and get rid of the signal sequence that is not present in the original fully
processed protein. Changing hydrophobic amino acids to charged amino acids both destroys
the ability of the sequence to act as a signal sequence and to become a membrane-spanning
sequence. So, mutating the first signal sequence so that it loses its function (A) will also cause
the second signal sequence to become an internal start-transfer sequence. In this case, howev-
er, the extreme amino terminus would remain on the cytoplasmic side of the membrane and
the mutated signal sequence would not be cleaved off because signal peptidase is found only
inside the ER (Figure A14–20B). Mutating every other signal sequence (B) so that it loses its
function would decrease the number of transmembrane regions and increase the size of the
internal loops between membrane-spanning regions (Figure A14–20C). Adding a new signal
sequence to the carboxyl terminus of the protein (C) will have no effect on the orientation of
the protein in the membrane, since the amino terminus will have been inserted before the
final signal is even translated.
deletion of first signal mutation in first signal mutation in every
other signal

ER lumen
C C * *

N C

*
N N

A14–20 (A) (B) (C)

Chapter 14: Intracellular Compartments and Transport 203

A14–21. A. Proteins are transported out of a cell via the secretory or exocytic pathway.
B. Fluid and macromolecules are transported into the cell via the endocytic pathway.
C. All proteins being transported out of the cell pass through the endoplasmic reticulum
and the Golgi apparatus.
D. Transport vesicles link organelles of the endomembrane system.
E. The formation of disulfide bonds in the endoplasmic reticulum stabilizes protein struc-
ture.

A14–22. B. An individual vesicle may contain more than one type of protein in its lumen (A), all of
which will contain the same sorting signal (or will lack specific sorting signals). Endocytic vesi-
cles (C) generally move away from the plasma membrane. The vesicle will not necessarily con-
tain the same lipid and protein composition as the donor organelle, since the vesicle is formed
from a selected subset of organelle membrane and contents from which it budded (D).

A14–23. B. COP proteins are thought to coat the vesicles used in ER to Golgi transport. Clathrin coats
form around vesicles budding from the Golgi and from the plasma membrane (A and C). GTP
hydrolysis is required for dynamin-induced pinching off of the vesicle bud (D). They do also
contain adaptin coat proteins (E).

A14–24. E. Adaptins are found only in clathrin-coated vesicles (A). They cannot hydrolyze GTP (B);
dynamin hydrolyzes GTP in clathrin-coated vesicles. Clathrin forms a basketlike network (C).
They are shed with the clathrin coat (D).

A14–25. (A) Clathrin-coated pits would form, but would not bud off. (B) Clathrin coats would not form
around the vesicle. Adaptins are required to link the clathrin to the membrane. (C) Without
clathrin, adaptins could still bind to the membrane, but no coat would form.

A14–26. D. vSNAREs are proteins on the cytoplasmic face of transport vesicles that are used in target
membrane recognition. vSNAREs are required for docking to the target membrane. Fusion of
the two membranes is distinct from docking and is thought to be catalyzed by a separate
group of proteins.

A14–27. D.

A14–28. D. Since the oligosaccharide chain is modified by enzymes in the ER and Golgi, most proteins
leaving the Golgi do not contain the 14–sugar oligosaccharide that was originally transferred
from dolichol to the protein. A few cytoplasmic proteins are glycosylated (A), albeit with a sin-
gle sugar residue. Sugars are transferred as a single branched, preassembled oligosaccharide
from dolichol to the protein (B). The oligossaccharide contains mannose, glucose, and
N-acetylglucosamine (C). Glycosylation occurs as soon as the first suitable Asn enters the
endoplasmic reticulum lumen (E).

A14–29. 1 = B; 2 = E; 3 = A; 4 = D; 5 = C.

A14–30. A = 3 (oligosaccharide protein transferase = ER ); B = 1 (galactose transferase = central Golgi

cisternae); C = 4 (SA transferase = trans Golgi network); D = 2 (GlcNAc transferase = cis Golgi
network). Proteins are modified in a stepwise fashion in the Golgi apparatus, with early steps
taking place in the cis Golgi, intermediate steps taking place in the central Golgi cisternae, and
late steps occurring in the trans Golgi network. If each enzyme produces the substrate for the
next step, then a mutant lacking the enzyme that catalyzes the addition of the first sugar will
be missing all of the sugars, a mutant lacking the enzyme that catalyzes the addition of the
second sugar will contain the first sugar but will lack the other three, and so on. By this logic,
204 Essential Cell Biology Test Bank

mannose and GlcNAc must be the first sugars added, additional GlcNAc the second, galactose
the third, and SA the last. Hence, the oligosaccharide protein transferase must be in the ER, the
GlcNAc transferase in the cis Golgi, the galactose transferase in the central Golgi, and the SA
transferase in the trans Golgi.

A14–31. A. New plasma membrane reaches the plasma membrane by the constitutive exocytosis path-
way.
B. New plasma membrane proteins reach the plasma membrane by the constitutive exocy-
tosis pathway.
C. Insulin is secreted from pancreatic cells by the regulated exocytosis pathway.
D. The interior of the trans Golgi network is acidic.
E. Proteins that are constitutively secreted do not aggregate in the trans Golgi network.

A14–32. C. Since ions do not have specific receptors, they are nonspecifically packaged into vesicles, so
we expect both types of vesicle to have similar concentrations of Ca2+ as the donor organelle,
the Golgi apparatus. tSNAREs mark the target membranes not the vesicles (A). The concentra-
tion of proteins in regulated vesicles is much higher due to the ability of the proteins packaged
into such vesicles to aggregate (B). Regulated vesicles require additional stimulation in order to
fuse with the plasma membrane in vivo, so we expect that these vesicles should also require
the addition of an activating fraction or removal of an inhibitory fraction in order to fuse in
vitro (D). Vesicles vary in size, but regulated secretory vesicles can be huge (relatively speak-
ing), as one might expect, since they are formed around aggregates of protein (E).

A14–33. C. Particles lacking surface molecules that are able to bind to macrophage receptors do not
stimulate phagocytosis and are therefore not engulfed; this mechanism prevents macrophages
from devouring the animal’s own cells. Pinocytosis and phagocytosis are forms of endocytosis,
not exocytosis (A). The rate of pinocytosis varies from cell type to cell type (B). Phagocytic vesi-
cles start out as pseudopodia that are extended around the particle to be engulfed (D). Both
pinocytic and phagocytic vesicles fuse with the endosome (E).

A14–34. E. Protease treatment will digest any LDL protein bound to the cell surface. If the LDL protein
is still present in the cell lysate after protease treatment, this indicates that it has been endocy-
tosed, since the protease cannot cross the membrane. The results from the “2 minute” experi-
ment show that both your cells and your sister’s cells take up LDL identically. Therefore, you
must have the same amount of LDL receptor as your sister (so B cannot be the case), and your
receptors must be able to bind LDL, form clathrin-coated pits and be internalized (so C and D
cannot be the case). The “40 minute” experiment shows that after this time, all the LDL taken
up by your sister’s cells has been degraded, as none is present, whether or not protease is
added to the cell sample. This is most likely due to the LDL having been transferred to the lyso-
some and hydrolyzed. LDL can still be found in your cells, on the other hand. The results from
your cells show that the LDL has not been degraded and instead has reappeared on the cell
surface, since it is once again sensitive to the added protease. By permanently blocking recep-
tors in this way it prevents efficient uptake of LDL from the blood; hence your tendency to
high blood cholesterol. Although there is the possibility that you could also have a defect in
your own LDL particles (not tested for), if that were your only defect (A), your cells and your
sister’s cells should behave identically at both time points.

A14–35. 1 recycled to the original membrane; 2 destroyed in the lysosome; 3 transcytosed across the
cell to a different membrane.
Chapter 14: Intracellular Compartments and Transport 205

A14–36. W = 3 (defect in mannose-6-phosphate receptor); X = 2 (defect in phosphotransferase); Y = 1

and Z = 1 (defect in lysosomal hydrolases). These will be defects in two different lysosomal acid
hydrolases. A cell that has no mannose-6-phosphate receptor will be able to make all the lyso-
somal hydrolases properly but will not be able to send them to the lysosome. Nor will it be able
to scavenge hydrolases from the external media. Hence, this cell line cannot be rescued by cul-
ture media that has had lysosomal hydrolases secreted into it and thus will not be rescued by
any of the media tested here. A cell line that has no phosphotransferase will be able to scav-
enge hydrolases from the external medium, but since all of the cell’s own hydrolases will lack
the mannose-6-phosphate tag, it will be rescued only by media from a cell line that is able to
make all of the hydrolases. Cell lines missing one hydrolase will be rescued by media from any
cell line that is able to secrete that hydrolase in a mannose-6-phosphate tagged form; in addi-
tion, media from cultures of cells missing a hydrolase will rescue any cell line with another
type of defect.