12.1 Cell Nucleus and Gene Expression Flashcards

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1
Q

What does a Centromere contain? Where does it bind

A

•Contains a tandemly repeated 171 bp DNA sequence called α satellite DNA
•Binds to specific proteins
E.g. CENP-A protein

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2
Q

Where are chromosomes localized in nucleus? specific chromo examples

A

Related to chromosome activity
-More active chromosomes are centrally located (less peripheral)
-Less active chromosomes are more peripheral
-Chromosome 19 has a greater density of protein coding genes
•more active,
•centrally located
-Chromo 18 peripheral, less dense

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3
Q

What’s the interaction between distant genes?

A
  • genes with common purpose move together
  • these genes are up regulate in response to estrogen
  • after estrogen there’s only 2 localized regions. coming closer together
  • genes on dif regions of the same/dif chromo can come together in the nun
  • may have common pool of proteins involved in the transcription process
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4
Q

mRNA processing machinery is localized where?

A
  • mRNA machinery is localized to small discrete sites or speckles
  • compartmentalized
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5
Q

What’s the nuclear matrix?

A
  • network of proteins very resistant to solubalization. This protein network will remain if you try to break own a cell. NB in dynamics of a cell in gene transcription.
  • distinct protein network
  • know it exists and has protein network that’s distinct
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6
Q

4 Charactereistics of the Nuclear Matrix

A
  • Insoluble in high salt and high detergent conditions
  • Composed of network of crisscrossing protein fibrils
  • Scaffold for protein structure
  • Anchor for machinery for transcription, RNA processing
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7
Q

Control of Gene Expression in Eukaryotes

A
  • Euk=highly specialized cells (liver, kidneys, etc.)
  • dif between specialized cells are what’s expressed and what isn’t
  • Cells become differentiated
  • Still retain complete genetic information
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8
Q

Dolly the sheep

A
  • Enucleated egg cells fused with cells derived from mammary gland (epithelial cells)
  • Reduced serum levels caused epithelial cells to enter quiescent state (G0) (via starvation) that “unmasked” unexpressed genes
  • Fused cells “activated” by brief electric pulse.
  • Egg implanted in surrogate sheep
  • Dolly proved that even cell has all the genetic material needed.
  • enuclulatted = cell with nucleus removed
  • Dolly’s Death: due to telomers. from 6 years old cell she already had shorter tellers and so that probably set her up for a faster death.

A somatic cell is a regular body cell, not an egg cell. The nucleus of the somatic cell was removed and put into an unfertilised egg cell. The process is called somatic cell nuclear transfer. The cell used as the donor for the cloning of Nam was taken from a mammary gland. The production of a healthy clone therefore proved that a cell taken from a specific part of the body could recreate a whole individual. She was cloned so she did not need any male cells to fertilize the egg and mature it. She only has one parent.

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9
Q

What are the 4 levels of control of Gene expresssion

A
  • Transcription Level
  • Processing Level
  • Translational level
  • Posttranslational
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10
Q

Transcription Level control

A

-Control over whether a specific gene is transcribed and how often.

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11
Q

Processing Level Control

A
  • How a primary transcript is spliced

- how mRNA is spiced into RNA (give you 2 dif protein products)

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12
Q

Translational level control

A
  • Control over whether a transcript is translated or not and if so, at what levels.
  • miRNAs play a role in controlling translation
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13
Q

Posttranslational Control

A
  • Mechanisms that regulate activity and stability of proteins
  • some structures are degraded by proteasomes.
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14
Q

Gene Expression Pattern in Mouse Ex.

A

looking at myogenic, muogenin promoter is fused to gene that expresses a dye, a pigment. Tells that myogenic is promoting expression in the black parts in somites (early muscle development)

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15
Q

Gene Expression of TCA-cycle Enzymes

A
  • TCA cycle enzyme is induced as the cells begin to metabolize ethanol and repressed when ethanol exhausted
  • (repressed because cells become fermutative, turned off, down regulate their TCA enzymes)
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16
Q

2 Transciptional Factors

A

General Transcription Factors

  • Bind at core promoter sites
  • Core promoter is normally at first 500bp, anything upstream is just promoter region

Specific Transcription Factors

  • Bind to specific regulatory sites in genes
  • Interact with other transcription factors
  • which genes are turned on in which cell types.
  • Regulatory site = DNA sequence
  • 2 domain = protein binding domain (binds other trasciption factors) and DNA binding (binds DNA?)
17
Q

Transcription Factor Interactions

A

Transcription factors bind to specific regions of DNA (gene) and also interact with other transcription factors.

18
Q

Structure of Transcription Factors

A
  • 2 Alpha helical structures bind to DNA
  • Binding depends on combination of van der Waals interactions, ionic bonds, and H-bonds between the amino acids and DNA.
  • 2 binding sites for each protein in the dimer
  • backbone of DNA is negative so protein is +
19
Q

the 3 main classes of trasciption factor:

A
  1. zinc finger motif
  2. helix loop motif
  3. Leucine Zipper Motif
20
Q

Zinc Finger

A
  • largest class of TF
  • cis and histone form the finger
  • Zinc ion coordinated between two cysteine and two histidine residues
  • common to have multiple fingers
21
Q

9ZFM:

A
  • motif made with zinc his and cis creates finger and inserts itself into DNA backbone)
  • first ZFP discovered
  • TFIIIA bound to DNA of 5S rRNA gene
  • Zinc finger motif transcription factor
  • 9 zinc fingers in TFIIIA
22
Q

Helix Loop Motif

A
  • Two alpha helical segments separated by an intervening loop. Domain is often preceded by a stretch of highly basic amino acids whose positively charged side chains contact the DNA and determine sequence specificity of the transcription factor. Always occur as dimers
  • 2 helices that form a loop.
  • 2 helix proefins dimerizing togehter to form an active TF
  • example is MyoD NB for differentiating into muscle cells
  • Myogenein is turned on my MyoD
  • Heterodimerization increases diversity of regulatory factors.
  • can either be homo or heterodimer
  • home = 2 identical proteins dimerizing together
  • hetero = 2 die pretiös dimerizing together
  • gert’s lots of diversity due to dif things dimerizing together
23
Q

Leucine Zipper Motif

A

-Transcription factors bind to specific regions of DNA (gene) and also interact with other transcription factors.
-3 class: they have a leucine a.a even 7th residue
made of alpha helix
called zipper because alpha helix zip together (dimerize together)
-each turn of an å-helix is 3.5 a.a so that the 7th is NB
their all (luc) on the same side are hydrophobic, they help to zip them together.