INTERACTIONS AMONG ORGANELLES

Organelles are involved in important cellular processes, such as the uptake and release of
material by cells, protein synthesis, and intracellular digestion. These various interactions
provide the basis for a functional approach to examine some dynamics of cell biology.

A. Uptake and release of material by cells

1. Endocytosis is the uptake (internalization) of material by cells. Endocytosis includes

pinocytosis, receptor-mediated endocytosis, and phagocytosis.
a. Pinocytosis (cell drinking) is the nonspecific (random) uptake of extracellular
fluid and material in solution into pinocytic vesicles.
b. Receptor-mediated endocytosis is the specific uptake of a substance (e.g., low-
density lipoproteins [LDLs] and protein hormones) by a cell that has a plasma membrane
receptor for that substance (which is termed a ligand). It involves the following
sequence of events (Figure 3.5):
(1) A ligand binds specifically to its receptors on the cell surface.
(2) Ligandreceptor complexes cluster into a clathrin-coated pit, which invaginates
and gives rise to a clathrin-coated vesicle containing the ligand.
(3) The cytoplasmic clathrin coat is rapidly lost, leaving an uncoated endocytic vesicle
containing the ligand.

t a b l e 3.
CLIc. Phagocytosis (cell eating) is the uptake of microorganisms, other cells, and
particulate matter (frequently of foreign origin) by a cell. Phagocytosis usually involves cell
surface receptors. It is characteristic of cellsparticularly macrophages that degrade
proteins and cellular debris and involves the following sequence of events:
(1) A macrophage binds via its Fc receptors to a bacterium coated with the antibody
immunoglobulin G (IgG) or via its C3b receptors to a complement-coated bacterium.
(2) Binding progresses until the plasma membrane completely envelops the bacterium,
forming a phagocytic vacuole.

2. Exocytosis is the release of material from the cell via fusion of a secretory granule
membrane
and the plasma membrane. It requires interaction of receptors in both the secretory
granule membrane and the plasma membrane as well as the coalescence (adherence and
joining) of the two phospholipid membrane bilayers. Exocytosis takes place in both regulated
and constitutive secretion.
a. Regulated (signal directed) secretion is the release, in response to an extracellular signal,
of proteins and other materials stored in the cell.
b. Constitutive secretion (default pathway) is the more or less continuous release of
material (e.g., collagen and plasma proteins) without any intermediate storage step. An
extracellular signal is not required for constitutive secretion.
3. Membrane recycling maintains a relatively constant plasma membrane surface area
following exocytosis. In this process, the secretory granule membrane added to the plasma
membrane surface during exocytosis is retrieved through endocytosis via clathrincoated
vesicles. This membrane is returned to the TGN via early endosomes for further recycling.
Figure 3.11 illustrates endocytic pathways used by cells.

B. Protein synthesis
1. Synthesis of membrane-packaged proteins involves translation of mRNAs encoding the
protein on polyribosomes at the surface of the RER, transport of the growing polypeptide
chain across the RER membrane and into the cisterna (lumen), and its processing within the
RER. These water-soluble proteins will bud from the RER and be transported in vesicles
either for transfer into the lumen (or interior) of another organelle or for secretion from the
cell.
a. A three-step process translates mRNA as follows: mRNA binds to the small subunit of a
ribosome that has three binding sites (A, P, and E) for tRNA molecules (Figure 3.12).

The tRNA anticodon sites base-pair with complementary codon sites in the mRNA, and
because only one particular type of the many tRNAs in a cell can base-pair with each
codon, it is the codon that determines which amino acid will be added to the peptide chain.
Once the start codon (AUG for methionine) is recognized and the initiator tRNA (bearing
methionine) is attached to the P site, the large ribosome subunit combines with the small
subunit, and protein synthesis begins. The next codon is recognized by an aminoacyl tRNA
bearing the proper amino acid, which then binds to the A site (1st step). Methionine at the P
site forms the first peptide bond with the incoming amino acid forming a dipeptide (2nd step).
The mRNA moves a distance of one codon (three nucleotides) through the small subunit, and
the spent initiator tRNA moves to the E site and is ejected, leaving the A site empty so that
a new aminoacyl tRNA can bind (3rd step). The A site then becomes occupied by an
aminoacyl tRNA bearing the next amino acid to be added, which forms a peptide bond with
the growing chain at the P site, and the initiator tRNA is ejected from the E site and the
process repeats over and over until the stop codon is reached and protein synthesis ceases.
The ribosome moves along the mRNA in the 5_ to 3_ direction using acylated tRNAs as
adapters to add each amino acid to the end of the growing peptide chain, which is always
located at the P site of the large subunit of the ribosome.
b. Transport of the newly formed peptide into the RER cisterna is thought to occur by a
mechanism described by the signal hypothesis as follows (Figure 3.13):
Chapter 3
FIGURE 3.11. Pathways used by receptors and ligands following endocytosis. (1) Recycling
of receptors to the same plasma membrane surface. (2) Transcytosis from one surface (e.g.,
basallateral) to another (e.g., apical). Transcytosis can occur in either direction, but separate
early endosome compartments near the domain of vesicle entry are used. (3) Degradation: If
not retrieved from either early endosome compartment, the ligands move to a common late
endosome, which subsequently becomes a lysosome, where degradation is completed.

FIGURE 3.12. A diagram illustrating protein synthesis

(1) mRNAs for secretory, membrane, and lysosomal proteins contain codons that
encode a signal sequence.
(2) When the signal sequence is formed on the ribosome, a signal recognition particle
(SRP) in the cytosol binds to it.
(3) Synthesis of the growing chain stops until the SRP facilitates the relocation of the
polysome to SRP receptors in the RER membrane.
(4) The large subunits of the ribosomes interact with ribosome receptor proteins,
which bind them to the RER membrane. The SRP detaches, and multisubunit protein
translocators form a pore across the RER membrane. Synthesis resumes, and
the newly formed polypeptide is threaded through the pore and into the RER cisterna
(lumen).
b. Posttranslational modification in the RER
(1) After the newly formed polypeptide enters the cisterna, a signal peptidase cleaves
the signal sequence from it.
(2) The polypeptide is glycosylated.
(3) Disulfide bonds form, converting the linear polypeptide into a globular form.
c. Protein transport from the RER to the cis Golgi (Figure 3.14)
(1) Transitional elements of the RER give rise to COP-II coatomercoated vesicles
containing newly synthesized protein.
(2) These vesicles move to the VTC where they deliver the protein.
(3) The VTC appears to be the first way station for the segregation of anterograde versus
retrograde transport in the secretory pathway. Either proteins move forward toward the cis
Golgi, or if they are RER-resident proteins that escaped from the RER, they are captured by a
specific membrane receptor protein and returned in COP-I coatomer-coated vesicles to the
RER along a microtubule guided pathway.

FIGURE 3.13. The signal hypothesis. The signal sequence of a newly formed secretory
polypeptide binds to an SRP that delivers the ribosomepeptideSRP complex to a receptor
on the RER. The SRP is recycled, and the polypeptide is translocated into the cisterna of the
RER, where a signal peptidase cleaves off the signal sequence.
FIGURE 3.14. The pathways of secretory proteins in separate compartments of the Golgi
complex. Proteins synthesized in the RER include secretory (_), membrane (/), and lysosomal
(o) proteins. These proteins bud off the transitional element of the RER via COP-coated
vesicles and enter the vesiculartubular cluster (VTC). From here they are transported to the
cis Golgi via COP-coated vesicles. Anterograde passage through the Golgi cisternae is either
via COP-coated vesicles or by cisternal maturation. Cisternal maturation coupled with
retrograde vesicular transport of Golgi enzymes is currently a favored view. Only COP-I
vesicles are thought to function in retrograde transport (from Golgi to VTC or RER), but both
COP-II and COP-I may function in anterograde transport. Not all proteins undergo all of the
chemical modifications (e.g., only lysosomal proteins undergo tagging with mannose 6-
phosphate). Final sorting occurs in the trans-Golgi network (TGN). Cytoplasm and
Organelles

d. Anterograde transport from the VTC to the cis Golgi is via COP-II coatomercoated
vesicles.
e. Movement of material anterograde among the Golgi subcompartments may occur by
cisternal maturation and/or by vesicular transport, as follows:
(1) Cisternae containing proteins may change in biochemical composition as they move intact
across the stack.
(2) COP-IIcoated vesicles may bud off one cisterna and fuse with the dilated rim of another
cisterna.
(3) Although both mechanisms have been observed, the precise way that anterograde
transport occurs across the Golgi stack of cisternae is unresolved.
(4) Retrograde vesicular transport occurs between Golgi cisternae and between the Golgi
and the VTC or RER via COP-Icoated vesicles.
f. Protein processing in the Golgi complex (Figure 3.14) occurs as proteins move from the
cis to the trans face of the Golgi complex through distinct cisternal subcompartments.
Protein processing may include the following events, each of which occurs in a different
cisternal subcompartment:
(1) Proteins targeted for lysosomes are tagged with mannose 6-phosphate in the cis cisterna.
(2) Mannose residues are removed in cis and medial cisternae.
(3) Some proteins undergo terminal glycosylation with sialic acid residues and galactose.
(4) Sulfation and phosphorylation of amino acid residues take place.
(5) A membrane similar in composition and thickness to the plasma membrane is acquired.
g. Sorting of proteins in TGN (Figure 3.14)
(1) Regulated secretory proteins are sorted from membrane and lysosomal proteins and
delivered via clathrin-coated vesicles to condensing vacuoles, in which removal of water via
ionic exchanges yields secretory granules.
(2) Lysosomal proteins are sorted into clathrin-coated regions of the TGN that have
receptors for mannose 6-phosphate and are delivered to late endosomes via clathrin-coated
vesicles.
(3) Plasma membrane proteins are sorted into coatomer-coated regions of the TGN and
delivered to the plasma membrane in COP-II coatomercoated vesicles.
2. Synthesis of transmembrane proteins also takes place on polyribosomes at the surface of
the RER, but rather than entering the lumen, the transfer process is halted (by a stoptransfer
sequence), and the transmembrane protein becomes anchored in the RER membrane. The
ultimate destination of this protein will be the RER membrane, the membrane of another
organelle, or the plasma membrane.
3. Synthesis of cytosolic proteins takes place on polyribosomes lying free in the cytosol and
is directed by mRNAs that lack signal codons. Such proteins (e.g., protein kinase and
hemoglobin) are released directly into the cytosol.

C. Intracellular digestion
1. Nonlysosomal digestion is the degradation of cytosolic constituents by mechanisms
outside of the vacuolar lysosomal pathway. The major site for the degradation of unwanted
proteins is the proteosome, a cylindrical complex of nonlysosomal proteases. Proteins
marked for destruction are enzymatically tagged with ubiquitin, which delivers them to the
proteosome, where they are broken down to small peptides.
2. Lysosomal digestion (Figure 3.8) is the degradation of material within various types of
lysosomes by lysosomal enzymes. Different lysosomal compartments are involved,
depending on the origin of the material to be degraded.
a. Heterophagy is the ingestion and degradation of foreign material taken into the cell by
receptor-mediated endocytosis or phagocytosis.
(1) Digestion of endocytosed ligands occurs in multivesicular bodies (Figure 3.8).
(2) Digestion of phagocytosed microorganisms and foreign particles begins and may be
completed in phagolysosomes.
b. Autophagy is the segregation of an organelle or other cell constituents within membranes
from the RER to form an autophagic vacuole (Figure 3.9), which is subsequently digested in
an autophagolysosome.
c. Crinophagy is the fusion of hormone secretory granules and lysosomes and their
subsequent
digestion. Crinophagy is used to remove excess numbers of secretory granules from the cell.

CLINICAL