exam 2 control of gene expression: regulation of mRNA levels Flashcards

1
Q

how do cells respond to environmental changes

A

cells must be able to control the timing and level of gene expression in a dynamic fashion

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2
Q

where does a major component of gene expression regulation occur at

A

the level of mRNA expression - the first place where regulation can occur

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3
Q

what levels can mRNA be regulated at

A

transcriptional initiation, elongation, splicing, nuclear export, degradation

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4
Q

where does the best understood mechanism of regulation occur at

A

transcriptional initiation, and involves combinatorial control by transcription factors

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5
Q

what key roles do calcium ions have in cell activities

A

muscle contraction, cell division, secretion, endocytosis, fertilization, synaptic transmission, metabolism, cell movement

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6
Q

where can gene expression be regulated at

A

essentially every step from transcription through protein activity/stability:

  • transcription from DNA to RNA
  • splicing, RNA tail, 5’-cap addition
  • where RNA goes in cytoplasm
  • mRNA degradation and control
  • translation steps and initiation
  • protein activity control
  • protein degradation
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7
Q

where does the primary regulation of genes occur at

A

the level of RNA transcription - matching RNA synthesis to expression requirements avoids being very energetically costly

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8
Q

what are gene regulatory proteins/transcription factors

A

sequence-specific DNA-binding proteins that play a key role in defining the level of transcription

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9
Q

what do transcription factors contain

A

one or more of a small set of well-characterized DNA-binding motifs

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10
Q

what can transcription factors bind to and read

A

the outside of the DNA helix and influence the binding or activity of RNA polymerase II

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11
Q

what are the two accesses DNA has to being able to see bases

A

major groove and minor groove

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12
Q

where do most TF factors bind

A

major group because there is more info in the major groove than minor groove

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13
Q

what is different about the major groove

A

it’s not symmetrical = you can tell where bases are and which strand you are examining

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14
Q

what do the major and minor groove represent

A

opposite faces of a base pair

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15
Q

what’s the difference between major and minor groove

A
  • each base on a strand can be distinguished in the major groove
  • only AT base-pairs and GC base-pairs can be distinguished in the minor groove
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16
Q

what can a DNA-binding protein interact with

A

specific base pairs without unzipping DNA

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17
Q

what can occur through hydrogen bonding

A

interactions between the gene regulatory protein and a base-pair can occur

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18
Q

how many contacts are made by a gene regulator protein with DNA

A

10-20

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19
Q

what is the helix-turn-helix DNA binding motif

A

two alpha helices connected by a short unstructured stretch (“turn”) - helices are held at a specific angle by interactions between the helices

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20
Q

what does the C-terminal recognition helix of the helix-turn-helix motif make

A

sequence-specific contacts in the major groove of DNA and bind to DNA as symmetric dimers

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21
Q

what are symmetric dimers

A

two proteins are structurally similar and bind as mirror images

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22
Q

what happens with symmetric dimers

A

recognition helices bind to “half-sites” separated by one turn of the DNA helix on the same face of DNA

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23
Q

what does heterodimerization allow for

A

expansion of sequences recognized without expanding number of proteins made

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24
Q

where does the major groove fit in

A

the alpha helix

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25
what is the homeodomain
a special case of helix-turn-helix motif
26
what is the structure of the homeodomain
a larger structure of helix-turn-helix plus third alpha helix
27
what does the conserved structure of the homeodomain suggest
all homeodomains are presented to DNA in the same fashion
28
what is the zinc fingers DNA binding motif
one or more zinc ions (prosthetic group) is coordinated by amino acid side groups
29
what does one subclass of zinc fingers use
2 cysteines and 2 histidines to coordinate zinc between an alpha heliz and a 2-strand antiparallel beta sheet
30
where are zinc fingers usually found
in tandem clusters with a DNA-binding protein
31
what does a series of zinc fingers allow for
sequence specificity because each finger recognizes one or two base pairs
32
what does the second subclass of zinc fingers use
coordinates 2 zinc ions using 4 cysteines for each | - one zinc ion stabilizes a recognition helix and one stabilizes a loop involved in dimerization
33
what do zinc fingers bind to
DNA as symmetric dimers, similar to helix-turn-helix proteins
34
what is the leucine zipper motif
alpha helix containing a hydrophobic surface on one side
35
what do hydrophobic forces do to the leucine zipper
keeps lucines away from water and drives binding of two subunits together
36
what do proteins bind DNA as
as a dimeric structure
37
what happens with leucine zippers
the helix from one subunit binds to the corresponding helix in the second subunit in a coiled-coil structure - hydrophobic interactions
38
what does an alpha helix do in a leucine zipper
serves as both the dimerization region and the DNA-binding region
39
what is the helix-loop-helix binding motif
similar to helix-turn-helix but with a longer loop of proteins that connect to helices
40
what happens with helix-loop-helix structure
a short alpha helix is connected to a longer alpha helix by a flexible loop, which allows one helix to fold back and pack against the other
41
what does the helix-loop-helix structure act as
both a dimerization interface and the DNA-binding region, like with leucine zipper
42
what can dimerization of DNA-binding proteins enhance
binding and specificity by increasing the contact area with DNA
43
what does heterodimerization increase
the range of sequences that can be recognized
44
how many different sequences could be recognized by dimerization of a TF that has 3 different pairing partners, A B and C
6 - hetero and homodimerization
45
what are the two types of gene regulatory regions transcription factors generally act at
promoter or enhancer
46
what is the promoter
the region where RNA polymerase and the general transcription factors assemble
47
where is the promoter located
a short distance upstream of the 5' end of the gene
48
what is specific about a promoter
it is gene-specific and orientation is important
49
what is the enhancer
an independent region outside promoter
50
where is the enhancer located
very far away from the promoter and may be upstream, downstream, or within the gene
51
what can the enhancer region not drive on its own
transcription, but can increase it from its corresponding promoter
52
what is not specific about enhancers
they are position and orientation INDEPENDENT and can work with a promoter of a different gene (heterologous)
53
how are eukaryotic gene regulatory regions different than prokaryotic
they're more complex
54
what is combinatorial control of expression
multiple gene regulatory proteins work together to control the rate of transcription
55
how do TFs work cooperatively
two activators - increase transcription synergistically
56
how do TFs work antagonistically
an activator vs a repressor
57
what can combined use of repressors and activators enable
sophisticated control of transcription in bacteria - activator is only present in absence of glucose
58
what can TFs help to unpack*****
chromatin, making the gene accessible to RNA polymerase and the initiation complex
59
what can TFs control*****
recruitment of RNA polymerase and/or general TFs to the promoter
60
what can TFs regulate*****
the switch from initiation to elongation
61
what can TFs help recruit*****
histone-modifying enzymes to change the local chromatin structure
62
what can TFs bend*****
DNA to allow long-distance interactions between gene regulatory regions
63
can transcription factors serve as activators or repressors
both
64
what can transcription activators direct
local alterations in chromatin structure
65
why are TF selectively activated
cannot have every TF turned on in cells at all times
66
where are TFs regulated at
level of gene transcription - if not, must be regulated post-transcriptionally
67
what are TFs activated by
phosphorylation - MAPK family helps with this in response to signals from cell-surface receptors