Bio2382b Midterm 2 Sunday March 12.

250700838 - 251153654 HSB 35

11:00 – 11:45 AM. 30 questions. 45 mins. 251154205 - 251198772 HSB 236
Please go to the correct room!! 251198784 - 251205690 HSB 240
251205708 - 251212964 NSC 1
There will be an additional “office hour”
Thursday March 2nd noon – 2pm.
251212974 - 251215154 NSC 7
251215197 - 251217211 NCB 114
All of this information is also posted on 251217222 - 251218964 NCB 117
the course Owl website. 251218987 - 251220757 SSC 2024
251220766 - 251223918 SSC 2028
251223926 - 251226392 SSC 2036
251226422 - 251229047 SSC 3022
251229048 - 251230216 UCC 41
251230217 - 251233649 UCC 56
251233706 - 251239884 UCC 146
251239893 - 251339059 WSC 55
Section 8, Actin in none-striated muscle.
Intermediate filaments, and cell adhesion.
Section 8a Actin and myosin in none-striated muscle
1. Understand the role of actin and myosin in; cytokinesis, smooth
muscle contraction, vesicle transport, cytoplasmic streaming, and cell
migration.
2. Be able to describe the actin and myosin structures that are formed in
a migrating cell (filipodia, lamellipodia, focal adhesions, stress fibers.
3. Understand the roles (and order) played by Cdc42, Rac and Rho in
the formation of the above structures.
4. Be able to describe the 4 steps of cell movement described in class.
 Actin and Myosin in Non-Skeletal Muscle Cells
5 examples – cytokinesis, smooth muscle, vesicle transport, cytoplasmic streaming, cell migration
Localization of myosin II during cytokinesis

17.34

Such actin/myosin organization is seen in contractile rings, circumferential

belts and stress fibres.
Smooth Muscle Myosin phosphorylation is a major regulator of
Contraction. smooth muscle contraction.

Smooth muscle lacks troponin and troopmyosin

Ca2+ (both external and released from SR) alters

myosin confirmation via calmodulin (CaM).

Phosphorylation of the regulatory light chain

(LC) allows conformational change and
contraction.

Much slower and more persistent contraction than

skeletal muscle.

Can be regulated by a variety of extracellular

17.35 signals.
Vesicle Transport.
Myosin-V-bound vesicles are
carried along actin filaments
Things to note:
Actin is capped by formin.

Transport of many types of vesicles.

Transport of mRNA.

Positioning of nucleus via MTs.

(interaction of actin and MTs).

Myosin V is inactive in the absence of

cargo.
 Myosin generates cytoplasmic streaming
In alga Nitella actin rings works with myosin to move vesicles etc. Helps with diffusion.

https://av.tib.eu/media/15043

17-37
 Cell Migration
Stress fibers associated
with cell adhesion are long
actin fibres which are
interacting with myosin
and other regulatory
molecules. These fibers are
contractile.

17-39
Chemotaxis
What Provides Directional Cues?
What does a cell need on its surface to respond?

What is the cell moving along?

Related to 17-45
4 steps of cell movement

17-38
 Cdc42, Rac, Rho,
actin and cell migration

Cdc42, Rac and Rho – all are “Rho proteins”

(members of Ras superfamily of small
GTPases)

-In GTP active forms can trigger

“effector proteins” 17-40,41
Cell migration involves coordinate regulation
of Cdc42, Rac and Rho.

17-42,43
Cdc42 important in polarity
 Cell migration involves coordinate regulation
of Cdc42, Rac and Rho.

Cdc42 important in polarity AND

coordinating actin and microtubules!

17-44

18.55
Section 8b Intermediate filaments
1. Understand the differences between intermediate filaments and action and tubulin
(no nucleotide, polarity, motors, less dynamic).

2. Remember the tissue specificity of keratin, vimentin, desmin, and neurofilaments.

3. Be able to explain how IFs and their binding proteins can support membranes.

4. Be able to describe the role played by intermediate filaments in the formation of

desmosomes and hemidesmosomes.
Intermediate filaments
- not globular
- no ATP or GTP needed
- no polarity
- no known motor proteins
- less dynamic
Tetramer is basic subunit

18-50

18-1
Main types of intermediate filaments

New table 18-2

Intermediate filament associated proteins
Only organizational proteins identified (no motors, caps, severing).
Plectin (green) cross-links IF vimentin (blue) and microtubules (red)

18-54
IF functions – membrane attachments, nucleus
IFs provide structural support for cell shape
eg. vimentin links to ankyrin at plasma membrane
Lamins A, B and C support the nuclear membrane.
B lamins are ubiquitous, (and linked via prenylation).
Cytoplasmic cytoskeletal protein
(actin MTs, IFs) linked to “nuclear
lamina” (layers) via intermediate
filament associated proteins
(contained in LINC complex)
18.53
 IFs are in dynamic state - proteins within filament are exchanged

18-51

Need to disassemble during mitosis (cyclin dependent kinase roles). N-terminal domain of
lamin A phosphorylated at serine - induces disassembly, and prevents reassembly. If serine
residue is mutated no disassembly. Balance of opposing action of kinases and phosphatases
crucial.
IF functions – plasma membrane
attachments.
IFs crucial to epithelial and other tissue
integrity.

keratin mutations
Transgenic mice carrying a mutant
keratin gene exhibit skin 18.51
blistering – weak skin integrity

IFs anchored to desmosomes and

hemidesmosomes
IF junctions - desmosomes and hemidesmosomes

many complex organizations – even for epithelial cells

Section 8c Cell Adhesion
Cell-cell and Cell-ECM interactions
1. Be able to explain the functions of GAP junction and tight junctions.

2. Understand the difference between homo- and heterophilic adhesion and give examples
of such molecules, structure, and functions.

3. Give examples of ECM proteins and functions.

4. Be able to explain the process of extravasation using the specific examples of cell-cell
and cell-ECM adhesion.
 Cell Adhesion
4 types -Tight junctions -Gap junctions -Cell-cell adhesions -Cell-ECM adhesions

20-1
 Gap Junctions
6 connexins form a connexon

Ca2+ regulated channel

Allow passage and

communication via small
molecules

20-21
 Tight Junctions

Membranes from adjacent cells form barrier.

Involves rows of occludin, claudin and JAM proteins in each cell.
20-17,18,20
 Cell adhesion molecules - CAMs

20-2
 Homophilic cell-cell adhesion – Ig superfamily and Cadherins
Ig superfamily of CAMs
Mediate Ca2+ independent homophilic cell adhesion - eg NCAM, ICAM

Major Cadherin molecules

Over 40 types of cadherins - homophilic adhesion.
Single transmembrane domain, and a cytosolic C-terminal tail – associated with cytoskeleton.
Ca2+ dependent.
Essential form holding cells in sheets
– eg E-cadherin (epidermal tissues),
– N-cadherin (nervous tissues).

20-3
Cadherins – Cell Adhesion
functions depend on cytoskeletal elements as well.
Most cell adhesion structures link to the cytoskeletal in
such a way as to allow cell signalling.
Cells “know” if they are linked to each other or not.

20-14 part
Cell-Cell vs
Cell-ECM

20-11 part
The ECM (extracellular matrix) is a complex mix of secreted molecules that
all cells interact with, and thus it regulates many cellular functions
TABLE 20-2: Functions of the Extracellular Matrix

1. Anchoring and surrounding cells to maintain solid-tissue three-dimensional architecture and define tissue boundaries

2. Determining the biomechanical properties (stiffness/elasticity, porosity, shape) of the extracellular environment

3. Controlling cellular polarity, survival, proliferation, differentiation, and fate (e.g., asymmetric division of stem cells; see Chapter 22), and
thus embryonic and neonatal development and adult function and responses to the environment and to disease

4. Inhibiting or facilitating cell migration (e.g., serving as either a harrier to movement or, conversely, as a “track” along which cells—or
portions of cells—can move)

5. Binding to and acting as a reservoir of growth factors; in some cases, the ECM (a) helps generate an extracellular concentration gradient
of the growth factor, (b) serves as a co-receptor for the growth factor, or (c) aids in proper binding of the growth factor to its receptor
(ECM component and growth factor jointly serve as a receptor’s combined ligand)
6. Activating cell surface signaling receptors

ECM proteins can be hydrophilic, (proteoglycans

absorb water and provide tissue resiliency)
large and structural, (collagens provide much
strength and support) and adhesive (fibronectin
and laminin recognized by special cell-surface
proteins, integrins, to allow cell migration).
 Integrin-Ligand Interactions
Heterodimeric transmembrane protein. α1β1 binds collagen
Different alpha and beta combinations α5β1 binds fibronectin
recognize specific substrates. α6β1 binds laminin
Integrins bind to core RGD amino acids,
but also require nearby synergy regions
for specificity.

fibronectin

20-33
 Integrins connect cells to substratum (ECM)
• Hetrodimer - Many alpha and beta combinations
• recognize specific substrates in ECM or on cell surface
• Bind RGD
Integrins can be in active and inactive states and transduce cell signals.

20-39,08
 Selectins – recognize oligosaccharides
• Participate in leukocyte extravasation (P-selectin)

20-42
 TABLE 20-3: Cell Junctions

Adhesion Principal CAMs or Adhesion Cytoskeletal Intracellular

Junction Function
Type Receptors Attachment Adapters

Anchoring junctions

1. Adherens junctions Cell-cell Cadherins Actin filaments Catenins, vinculin Shape, tension, signaling, force
transmission
2. Desmosomes Cell-cell Desmosomal Intermediate Plakoglobin, plakophilins, Strength, durability, signaling
cadherins filaments
desmoplakins
3. Hemidesmosomes Cell- Integrin (α6β4) Intermediate Plectin, dystonin/ BPAG1 Shape, rigidity, signaling
matrix filaments

4. Focal contacts, fibrillar, and 3-D Cell- Integrins Actin filaments Talin, kindlin, paxillin, vinculin Shape, signaling, force transmission, cell
matrix
adhesions movement
Tight junctions Cell-cell Occludin, claudins, JAMs Actin filaments ZO-1,2,3, PAR3, cingulin Controlling solute flow, cell polarity,
signaling
Gap junctions Cell-cell Connexins, innexins, pannexins Via adapters to other ZO-1,2,3 Communication, small-molecule
junctions transport between cells

Plasmodesmata (plants only) Cell-cell Undefined Actin filaments NET1A Communication, molecule transport
between cells