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tru_cell > module 5 > Flashcards

module 5 Flashcards

1
Q

cytoskeleton is composed of

A
  1. microtubules
  2. actin filaments (microfilaments)
  3. intermediate filaments
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2
Q

microtubules

A

In epithelial and neuron cells:

  • Support and organelle transport

In dividing cell:

  • Form mitotic spindle required for chromosome segregation

Structural characteristics:

  • Long
  • Hollow
  • Unbranched
  • Composed of tubulin

found in nearly all euk cell

assembled from tubulin

polarity: (-) end in the middle of the cell, (+) towards the edges of the cell

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3
Q

actin filaments

A
  • Aka microfilaments
  • Support of microvilli of epithelial cell
  • Part of motile machinery in neuronal elongation and cell division
  • Structural characteristics:
  • Solid
  • Thin structures
  • Organized into branched networks
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4
Q

Intermediate filaments:

A
  • Structural support
  • Structural characteristics:
  • Tough
  • Ropelike
  • Composed of variety of related proteins
  • no polarity
  • include neurofilaments, major component of structural framework supporting neurons
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5
Q

Protofilaments:

A
  • Globular proteind arranged in longitudinal rows
  • Aligned parallel to the long axis of the tubule
  • Held by noncovalent interactions
  • Assembled from dimeric blocks of one alpha and 1 beta tubulin
  • Assymetrical: alpha on one side and beta on the other
  • All protofilaments have same polarity:
  • Important in growth of microtubules and their ability to participate in directed mechanical activities
    • end is beta
  • End(-) is alpha
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6
Q

microtubule associated proteins function

A
  • To increase the stability of microtubules
  • To alter microtubules rigidity
  • Influence the rate of microtubule assembly
  • Attach microtubules to one another maintaining parallel alignment
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7
Q

functions of microtubules

A
  • Support of the cell
  • Ciliary and flagellar motion
  • Movement of material b/ cell body and axon terminals
  • Mitosis/meiosis
  • Intracellular organization etc.
  • Structural support and organizer:
    • Distribution of microtubules helps to determine the shape of the cell
  • Influence on formation of cell wall
  • Maintaining the internal organization of the cells
  • Intracellular mobility:
    • Transport of material from one membrane compartment to another
    • Movement of vesicles along the axon in both directions
  • Serve as tracks for variety of motor proteins
    • Associated w/ cytoskeleton convert chemical E into mech E that is used for cellular movement
    • Move unidirectionally along their track in stepwise manner
    • Microtubules moves in the direction opposite to the direction that the motor is stepping
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8
Q

Importance of beta tubulin binding of GTP and subsequent hydrolysis

A
  • Beta tubulin is a GTPase
  • GDP after the hydrolysis remains bound to the assembled polymer
  • During disassembly: GDP->GTP
  • Presence of cap of tubulin-GTP dimer at + end favors addition of more subunits and the growth of microtubule
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9
Q

motor proteins associated w/ microtubules

A
  1. kinesin
  2. cytoplasmic dynein
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10
Q

kinesin

A
  • motor protein associted w/ microtubules
  • Structure:
    • Tetramer constructed from two identical heavy and two identical light chains
    • Pair of globular heads
      • Bind microtubule
      • ATP-hydrolysing engine
    • Neck
    • Stalk
      • Where H and L chains form DH
      • Binds cargo to be hauled
    • Tail
  • Moves towards + side of the tubule
  • Moves along a single protofilament of microtubule proportional to [ATP]
  • Moves via “hand-over-hand” mechanism
  • Movement is processive:
    • One protiens moves along individual microtubule for long distance
  • Independent, long-distance transport of small cargo
  • Important function during cell division
  • Force-generating agents for movement of the transport vesicles (tend to move in outward direction
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11
Q

cytoplasmic dynein

A
  • Structure:
    • Two identical heavy chains:
      • Large globular head:
        • Force generating engine
      • Elongated stalk:
        • Microtubule binding site
      • Tail:
        • Binds intermediate and light chains
    • Variety of intermediate and light chains
  • Moves towards (-) end
  • Functions:
    • Positioning the spindle and moving chromosomes during mitosis
    • Positioning centrosome and Golgi and moving organelles, vesicles and particles
    • In nerve cells:
      • Retrograde movement of organelles
      • Anterograde movement of microtubules
    • Retrograge movement towards the center of the cell
    • Responsible for movement of cilia and flagella
  • Requires intervening adaptor - dynactin
    • Increases processivity of dynein
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12
Q

dynactin

A
    • intervening adaptor for dynein
  • increases processivity of dynein
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13
Q

Functions of MTOCs:

A
  • They are variety of specialized structures that initate microtubule nucleation and organize microtubules
  • Control # of microtubules, their polarity, # of protofilaments that make up their walls and the time and location of their assembly
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14
Q

What are 2 different MTOC:

A

Centrosome:

  • In animal cells
  • Place from which newly assembled microtubules grow in all directions

Basal bodies:

  • Where microtubules of cilium and flagellum originate
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15
Q

How cilia and flagella can undergo their bending movements:

A
  • Dynein arms act as swinging cross-bridges that generate the force required for movement
    • Steps:
      • Dynein is tighly anchored to outer surface of tubule A and its globular heads point towards B tubule
      • Dynein arms anchored along tubule A of lower doublet attach to binding sites on tubule B of upper doublet
      • Power stroke: lower doublet slides towards basal end of upper doublet
      • Dynein arms detached from tubule B
      • Arms have reattached to the upper doublet so that another cycle can begin
  • Sliding-microtubule mechanism of ciliary or flagellar motility:
    • When cilium is straight: all outer doublets end at the same level
    • Cilium bending occurs when doublets on the inner side of the bend slide beyond those on the outer
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16
Q

Sliding-microtubule mechanism of ciliary or flagellar motility:

A
  • When cilium is straight: all outer doublets end at the same level
  • Cilium bending occurs when doublets on the inner side of the bend slide beyond those on the outer
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17
Q

distinct characteristics of intermediate filaments

A
  • Chemically heterogeneous
  • Assembled in tetramers formed by two antiparallel dimers
  • Lack polarity
  • Resist tensile forces
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18
Q

Role in IF in neurons:

A
  • IF include neurofilaments - major component of the structural framework supporting neurons
  • As the axon increases t=in diameter, the neurofilaments provide support
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19
Q

8 actin binding proteins and their functions

A

Nucleating proteins:

  • Form template to which actin monomers can be added

Monomer-sequestering proteins:

  • Bind to acti monomers and prevent them from polymerizing

End-blocking proteins:

  • Reglate the length of actin

Monomer-polymerizing proteins:

  • Stimulate actin polymerization during cell locomotion

Actin filament depolymerizing proteins :

  • Enhance depolymerization

Cross-linking proteins:

  • Promote formation of loose networks of filaments

Fimalent severing proteins:

  • Decrease the viscosity of the cytoplasm

Membrane-binding proteins:

  • Bind to the plasma membrane and allow it to protrude or invaginate from the cell
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20
Q

Nucleating proteins:

A
  • Actin binding proteins
  • form template to which actin monomer can be added
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21
Q

monomer sequeatering proteins

A

Actin binding proteins

binding actin monomers and prevents them from polymerizing

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22
Q

end-blockin proteins

A

Actin binding proteins

regulate length of actin

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23
Q

monomer polymerizing proteins

A

Actin binding proteins

stimulate actin polymerization during cell locomotion

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24
Q

actin filament depolymerizing proteins

A

Actin binding proteins

enhance depolymerization

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25
Q

cross-linking proteins

A

Actin binding proteins

promote formation of loose networks of filaments

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26
Q

filament serving proteins

A

Actin binding proteins

decrease viscosit of cytoplasm

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27
Q

Cellular activities involving motility of nonmuscle cells in which actin filaments are involved:

A

Cytokinesis

Phagocytosis

Cytoplasmic streaming

Vesicle trafficking

Cell-substratum interactions

Cell locomotion

Axonal growth

Chnges in cell shape

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28
Q

Actin filament assembly/disassembly:

A
  • ATP is important
  • Barded end incorporates monomer at faster rate
  • Barded and pointed ends require different [ATP-actin monomer] to elongate:
    • Barded needs less
  • Steps:
    • Add preformed actin filaments to the solution of actin in the presence of ATP
    • If [ATP] is high, actin is added at both ends
    • Addition continues only at the barded end because [ATP] is only high enough for addition at that end
    • Addition continues at the barded end, but loss of the monomer occurs at the point end to respond to the decrease in free monomers
    • Addition/removal rates change to maintain free monomer concentration stable
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29
Q

general structure of myosin

A
  • Head:
    • Contains:
      • Site that binds an actin filaments
      • Site that hydrolyses ATP to drive the mysosin motor
  • Tail:
    • Divergent and different in various types
  • Variety of light chains
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30
Q

Conventional (type II) myosin

A
  • Primarily motors for muscle contraction
  • Can be found in non-muscle cells:
  • Move towards barded end
  • Function in non-muscle:
    • Split the cell into two
    • Generate tension at focal adhesion
    • Cell migration
  • Ends of tails point towards the center of the filament and the globular heads point away from the center
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31
Q

Unconventional myosin:

A
  • Subdivided into 17 different classes
  • Contains single head
  • Unable to assemble into filaments in vitro
  • Can exert tension on plasma membrane
  • Unable to form filament
  • Operate as inidivual proteins
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32
Q

Stages of the cell cycle:

A
  • Interphase:
    • G1:
      • Cell grows and carries out normal metabolism
      • Organelles duplication
    • S phase:
      • DNA replication
      • Chromosome duplication
    • G2:
      • Cell growth
      • Preparation for mitosis
  • Mitosis:
    • PMAT:
      • Duplicated chromosomes are separated into 2 nuclei
      • cytokinesis:
        • Division of the entire cell into 2 daughter

Cell cycles vary among different types of cell depending on the cell type and environmental conditions

33
Q

important regulation stage (check points) of cell cycle

A

@ end of G1:

  • First transition pt START
  • If pass-> commited to the DNA replication
  • Uses Cdk

@ end of G2:

  • Uses mitotic cyclins

Middle of mitosis:

  • Determines if they’ll complete cell division of reenter G1 of the next cycle
34
Q

checkpoint control

A
  • Ensures that DNA is not damaged
  • Stop the progress of cell cycle if:
    • DNA is damaged
    • Critical processes have not been properly completed
  • Activated by the system of sensors that recognize DNA damage or cellular abnormalities
  • Cell stops its progress at one of those check points via Cdk inhibitor
35
Q

G0 stage:

A
  • Cells that are arrested in the state of no cell division (temporarily or permanent) preceding the initiation of DNA synthesis
  • Ex. nerve cells, muscle cells, RBC (b/c highly specialized and lost the ability to divide)
36
Q

How is the activity of cyclin-dependent kinases regulated:

A
  • Cyclin dependent kinases are enzymes that phosphorylate other protein and regulate cell cycle
  • Regulated by:
    • Presence of specific cyclins:
      • Required subunits for specific Cdk activities
    • Levels of cyclins that vary throughout the cell cycle partially controlling the activity of Cdks and secondarilly controlling the activities of the substrates phosphorylated by those specific Cdks
    • Other kinases, which phosphorylate them result in activation/deactivation depending on state of phosphorylation
    • Phosphatases dephosphorylate them
      • May inhibit/activate some Cdks
    • Subcellular localization may separate Cdks from their substrates or create microenvi for them
    • CDK inhibitors
    • Cdks can be targeted for degradation (ex. by ubiquitin-proteasome pathway)
37
Q

Prophase

A
  • Chromosomes condense
  • Formation of compact mitotic chromosomes
  • Chromosomes composed of two chromatids attached together at centromere
  • Golgi and ER fragment
  • Nuclear envelope dissapeares
38
Q

prometaphase

A

Chromosomal microtubules attach to the kinetochores pf chromosomes

Chromosomes are moved to spindle equator

39
Q

Metaphase:

A

Chromosomes alligned at metaphase plate

Chromosomes attached to both poles by the microtubules

40
Q

anaphase

A

Centromere splits

Chromatids separate

Chromosomes move to the opposite spindle poles

Spindles poles move further apart

41
Q

Telophase:

A

Chromosomes cluster at opposite spindle poles

Chromosomes become dispersed

Nuclearenvelope assembles around chromosome cluster

Golgi and ER reform

Daughter cells form by cytokinesis

42
Q

centromeres

A
  • Primary constrictions in chromosomes
  • Residence of highly repeated DNA sequence
  • Serves as binding site for specific proteins
  • Has kinetochore:
43
Q

kinetochore

A
  • On the outer surface of centromere of each chromatid
  • Proteinaceous
  • Button-like
  • Function:
    • Residence of motor proteins involved in the chromosome motility:
      • Dynein
      • CENP-E
      • Depolymerase
    • Site of attachment of chromosome to the dynamic microtubules of mitotic spindle
    • Key component in the signalling pathway of an important mitotic checkpoint
44
Q

centrosome

A

initiate microtubule assembly that is responsible for formation of mitotic spindle

45
Q

Tubulin flux

A
  • Net addition of tubulin at + end (@ kinetochore side)
  • Net loss of tubulin @ - end
  • Subunits move along the chromosomal microtubules from kinetochore towards the pole
  • happens during metaphase
46
Q

anaphase promoting complex (APC)

A
  • Adds ubiquitin to proteins at different stages of the cell cycle targeting them for proteolysis
  • Adaptor protein:
    • Determines which proteins serve as APC substrates
    • APC-Cdc20:
    • APC-Cdh1:1
  • The importance of targeted proteasome-mediated proteolysis in progression thru mitosis:
    • Completion of mitosis requires the cessation of activity of Cdk1
47
Q

Cytokinesis:

A
  • In animal cells forms a furrow that moves inwards towards the center of the cell
  • In plants: form cell plate:
    • Secretory vesicles from Golgi align along the equatorial plane and begin to fuse with one another
    • Membrane of the vesicle become plasma membrnae
    • Content of the vesicle forms the cell plate separting cells
  • Role of actin filaments:
    • Becomes assembled in a ring at the cell equator
    • Contraction of the ring requires action of myosin -> formation of furrow that splits the cell in two
    • Are concentrated in a circular equatorial band within cleavage furrows
48
Q

purpose of meiosis

A

Reduce # chromosomes

Increase genetic variability

49
Q

Prophase I

A

formation of tetrads (synapsis b/ homologous chromosomes)

cross-over @ chiasmata

movements of tetrads towards the metaphase plate begins

50
Q

metaphase I

A
  • Two homologous chromosomes of tetra are connected to the spindle fibers from opposite pole
  • Sister chromatids are connected to the microtubules from the same spindle
  • Side-by-side arrangement of kinetochores
51
Q

anaphase I and telophase I

A

Anaphase I:

  • Homologous chromosomes separate
  • Dissolution of chiasmata by proteolytic cleavage of cohesin

Telophase I:

  • Chromosome dispersion, but don’t reach very extended states
  • Nuclear envelope may/not reform fully
52
Q

metaphase II

A

Kinetochores of sisters chromatids face opposite pole and are atached to the opposite sets of chromosomal spindle

53
Q

anaphase II

A

separation of sister chromatids

54
Q

how do nuclei formed in mitosis and meiosis differ from one another

A

In mitosis:

  • Pait of homologous chromosomes
  • Genetically identical to the mother cell

In meiosis:

  • Contain one chromosomes from each pair of homologous chromosomes contained by parental cell
  • Have the potential to be genetically different from parent chromosomes
55
Q

Signal transduction:

A

Process in which info carried by extracellular messengers is translated into changes that occur inside the cell

56
Q

G-protein:

A
  • GTP-binding proteins
  • Associated with:
    • Vesicle budding
    • Microtubule dynamics
    • Protein synthesis
    • Nucleoplasmic transport
  • Recognized by G protein-coupled receptos (GPCRs) that are composd of transmembrane alpha helices:
    • Capable of binding a large variety of ligands
  • Active when bound to GTP

Inactive when bound to GDP

57
Q

Mechanism of receptor-mediated activation of effectors by G proteins

A
  • Ligand binds to the receptor
  • Alters receptor conformation -> increases affinity for G protein
  • G protein alpha: GDP out, GTP in
  • G protein alpha dissociates from beta and gamma
  • Alpha-GTP binds to the effector (adenyl cyclase)
  • Activation of adenyl cyclase
  • Beta and gamma G protein bind to adenyl cyclase
  • Activated adenyl cyclase: ATP->cAMP
  • GTPase hydrolyses GTP->GDP and deactivates alpha G protein
  • G alpha reassociates w/ G beta and gamma
  • Adenyl cyclase stops its activity
  • GRK ( G protein-coupled receptor kinase) phosphorylates receptor with use of ATP
  • Arrestin binds to the receptor inhibiting it from activating more G proteins
  • Receptor-arrestin taken up by endocytosis
58
Q

Protein tyr kinase:

A

Involved in:

  • Regulation of cell growth
  • Cell division
  • Cell differentiation
  • Cell survival
  • Attachment to extracellular matrix
  • Migration of the cells

Expression of mutant protein tyrosin kinase can’t be regulated and leads to CA

59
Q

Necrosis:

A

Less orderly than apoptosis

Swelling of both cell and its internal membranous organelles

Membrane breaks

Leakage of cell content

Inflammation

60
Q

apoptosis

A

Neat abd orderly process:

  • Overall shrinkage in volume of cell and nucleus
  • Loss of adhesion to neighboring cell
  • Formation of blebs
  • Dissection of chromatin into small fragments
  • Rapid phagocytosis
61
Q

Cell signaling:

A
  • Process by which info is relayed across the plasma membrane to cell interior
  • Often this info is transmitted to the nucleus
  • Include:
    • Recognition of the stimulus
    • Transfer of signal across the plasma membrnae
    • Transmission of signal to specific effector
    • Cessation of the response
62
Q

Second messanger

A
  • Substance that is released into the cell as the result of binding of the 1st messanger (hormone or ligand) to a receptor outside the cell
  • Include:
    • cAMP
    • IP3
    • Ca2+
    • DAG
    • Nitric oxide
63
Q

Ras:

A
  • Is a G protein that is part of many pathway controlling cell growth
  • Needs to be quickly turned off after activation
  • In oncogenes, changes to Ras causes ot to be permanently on ->uncontrolled growth w/t differentiation ->tumor
  • Pathways are activated when growth factor bind to the extracellular domain of its receptor
64
Q

How do the properties of CA cells manifest themselves in culture:

A

Grow in multilayered clumps

Continue to grow regardless the presence/absence of exogenous growth factors

Unable to respond to inhibitory signals from neighboring cells

65
Q

What agents are carcinogenic:

A
  • Chemical mutagens
  • UV radiatino (forms thymidine dimers)
  • DNA- and RNA-tumor viruses:
    • Effects are more complicated
    • Some can induce transformation b/c they have appropriated cellular genes that can hinder cells’ growth control function
  • Inherited mutation
  • Diet:
    • Can increase or decrease risk of CA
66
Q

How does CA causing mutation arise?

A

From DNA damage caused by normal metabolic rnx that the cell unable to repair

67
Q

Basic properties of CA cells:

A
  • Malignant tumors tend to metastasize
  • CA cells have lost their growth control, and have following growth properties:
    • Grow in multilayered clumps
    • Continue to grow regardless the presence/absence of exogenous growth factors
    • Unable to respond to inhibitory signals from neighboring cells
  • CA cell often have highly aberrant chromosomes (aneuploidy), while healthy cells have a pair of identical chromosomes
    *
68
Q

genes involved in carcinogenesis

A

tumor suppressor genes

oncogenes

proto-oncogenes

69
Q

Tumor suppressor genes:

A
  • Act as cell’s brakes
  • Encode proteins that restrain cell growth and prevent cells from becoming malignant
  • If copies of the mutatte tumor suppressor genes on both homologous chromosomes -> loss of growth control
  • Loss-of-function mutation on both homologous->CA
70
Q

oncogenes

A
  • Encode proteins that promote the loss of growth control and the conversion of cell to a malignant state
  • Accelerators of cell proliferation
  • Lead to genetic instability
  • Prevent cell from apoptosis
  • Promote metastasis
  • Act dominantly (only need mutant copy on one of the homologous
71
Q

protoonco genes

A
  • Have the potential to subvert cell own activities and push the cell towards the malignant state
  • Encode proteins that have various function in cell normla activities
  • Can be converted into the oncogenes:
    • Mutation that will alter the properties of the gene product
    • Multiple duplication -> gene amplification -> excess products
    • Chromosome rearrangement ->altered gene expression
  • After gain-of-function mutation->oncogene
72
Q

mutations in RB genes that can lead to retinoblastoma

A
  • RB gene is ex. of tumor suppressor gene
  • Nonfamiliar ex of disease:
    • Sporadic cases
    • Person begins life w/ normal RB genes in zygote
    • Retinoblastoma occurs only in whom a given retinal cell accumulates independent mutation in both alleles of the gene
  • Familiar ex of disease:
    • Ex inherited
    • Abnormal allele in the zygote of the RB gene (usually present as a deletion)
    • All cells of the retina have at least one unfunctional RB gene
    • If the other gene is inactivated (ex. point mutation)->CA
73
Q

Types of proteins encoded by proto-oncogenes:

A

Receptor for growth factors

Protein kinases and proteins that activate proteins kinass

Proteins that regulate cell cycle

Transcription factors

Proteins that modify chromatin

Metabolic enzymes

Proteins that inhibit apoptosis

74
Q

Several families of oncogenes encode:

A

mutant forms of cytoplasmic protein kinases

Receptor protein kinases

GTP-binding proteins

75
Q

Benign tumour and malignant tumour

A
  • A benign tumour is composed of cancer cells that lack the capability to invade normal tissues or metastasize to distant cells
  • a malignant tumour tends to metastasize.
76
Q

Tumour-suppressor gene and oncogene

A
  • A tumour-suppressor gene encodes proteins that restrain cell growth and prevents cells from becoming malignant,
  • an oncogene encodes proteins that promote the loss of growth control and the conversion of a cell to a malignant state.
77
Q

proto oncogene vs oncogene

A
  • Proto-oncogenes are a variety of genes that have the potential to subvert the cells own activities, and push the cell toward a malignant state. They can be converted into oncogenes.
  • An oncogene encodes proteins that promote the loss of growth control and the conversion of a cell to a malignant state.
78
Q

The genes involved in carcinogenesis constitute a specific subset of the genome whose products are involved in distinct cellular activities. List three of these cellular activities.

A

Progression of a cell through the cell cycle

Adhesion of a cell to its neighbours

Apoptosis

Repair of DNA damage

Cell-to-cell communication

79
Q

What are the mechanisms by which proto-oncogenes are converted to oncogenes?

A
  • A gene mutation alters the structure and function of the encoded protein.
  • A mutation in a nearby regulatory sequence alters the expression of the gene.
  • A rearrangement of DNA brings a new DNA segment closer to the proto-oncogene, which results in an inappropriate transcription of the proto-oncogene by the promoter of the transposed gene.

tru_cell (6 decks)

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