Gene regulation Flashcards

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1
Q

all cells in an organism have the same …

A

…DNA sequence

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2
Q

regardless of their different morphology all cell types in the human body have…

A

…the same complement of DNA (with a couple of exceptions)

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3
Q

different cell types make…

A

…different proteins – yet proteins are the product of genes

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4
Q

proteins can be visualised by …

A

…electrophoresis

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5
Q

proteins can be separated by…

A

…electrophoresis as with DNA

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6
Q

proteins have different…

A

…charges (think of the different amino acids they are made up of)

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7
Q

Proteins have different charges (think of the different amino acids they are made up of) – hence we bind…

A

…SDS molecules to protein extracts which give proteins a uniform negative charge – an electric current separates them

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8
Q

same protein in both tissues is a…

A

…housekeeping gene

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9
Q

some genes are turned on in …

A

…all cells.

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10
Q

some genes are turned on in all cells

these are known as …

A

…housekeeping genes.

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11
Q

Housekeeping genes are involved in…

A

…processes that are fundamental to all cells – e.g. glycolysis, DNA replication, cell division control, RNA polymerases, ribosomal proteins etc

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12
Q

other genes are specific to…

A

…the individual type of cell

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13
Q

other genes are specific to the individual type of cell – these include genes such as those that …

A

…encode insulin (only expressed in β-cells of the pancreas) or CD4 which is expressed only in certain cells of the immune system

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14
Q

if all cells produced the same complement of proteins then they …

A

…could not differentiate

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15
Q

multicellular organisms depend upon…

A

…different functions being allocated to different cell types

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16
Q

multicellular organisms depend upon different functions being allocated to different cell types – therefore if all had the same protein complement they …

A

…could not do this

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17
Q
  • If all cells produced the same complement of proteins then they could not differentiate.
  • Multicellular organisms depend upon different functions being allocated to different cell types – therefore if all had the same protein complement they could not do this
  • So there is a need for…
A

… tissue specific gene expression (proteins are encoded by genes)

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18
Q
  • If all cells produced the same complement of proteins then they could not differentiate.
  • Multicellular organisms depend upon different functions being allocated to different cell types – therefore if all had the same protein complement they could not do this
  • So there is a need for tissue specific gene expression (proteins are encoded by genes).
  • So multicellular organisms have …
A

…spatial regulation of gene expression

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19
Q

expression of genes can be visualised in many ways – the following experiments use a …

A

…‘reporter gene’ (gives a colour change) fused to the promoter of the gene being studied – introduced using ‘genetic modification’ or transformation

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20
Q

the gene contains the sequence that will eventually be translated into …

A

… a protein

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21
Q

the gene contains the sequence that will eventually be translated into a protein

but adjacent to this lies a region of DNA called…

A

…the promoter

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22
Q

the promoter is involved in…

A

…deciding when and where a gene is expressed.

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23
Q

The promoter consists of a DNA sequence which…

A

…other proteins bind to and affect recruitment of RNA polymerase

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24
Q

the pictures of gene expression on the previous slide and following slides are obtained by …

A

…putting a reporter gene onto the promoter to see when and where the gene is expressed.

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25
Q

the reporter gene’s product gives …

A

…a colour when incubated with its substrate or glows.

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26
Q

the reporter gene’s product gives a colour when incubated with its substrate or glows

this is then transformed into an …

A

…animal or plant – genetic engineering

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27
Q

multicellular organisms go through a process of development from a single cell to the…

A

…complex multicellular adult organism

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28
Q

at different times certain genes may need to be switched on in order for organs to…

A

…develop correctly

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29
Q

if genes are not switched on in the right place and time then development will be …

A

… aberrant

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30
Q

multicellular organisms go through a process of development from a single cell to the complex multicellular adult organism

at different times certain genes may need to be switched on in order for organs to develop correctly – if genes are not switched on in the right place and time then development will be aberrant

so multicellular organisms have …

A

…temporal regulation of gene expression

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31
Q

cells are constantly bombarded by signals from the environment such as…

A

…light, temperature, pathogens, toxins, nutrients, signals from other cells

Causes changes in gene expression

32
Q

genes are regulated at various points in …

A

…transcription/translation

33
Q

the promoter of the tryptophan biosynthesis genes lies …

A

…upstream of the gene (i.e. before the transcription start point)

34
Q

it is the binding of transcription factors to the…

A

…operator region of the promoter that determines whether a gene is on or off

35
Q

proteins bind to the …

A

…minor groove or major groove of DNA

36
Q

hydrogen bonds form between amino acids on the …

A

…protein and the bases in DNA like below

37
Q

proteins binding to the promoter can either …

A

…repress transcription or activate transcription

38
Q

in the above example the tryptophan synthesis genes are turned off when …

A

…a repressor protein that is bound to tryptophan binds to an ‘operator’ region within the promoter in this way when tryptophan is present.

39
Q

binding of tryptophan to the repressor protein causes a …

A

…conformational change in the repressor protein.

40
Q

binding of tryptophan to the repressor protein causes a conformational change in the repressor protein.

as the protein changes shape it is able to…

A

…bind to the promoter and repress transcription – without tryptophan the parts of the protein involved in binding were unable to bind DNA

41
Q

the binding of a protein to the promoter helps to recruit …

A

…RNA polymerase to the promoter and so activates transcription

– an example of both activation and repression will be seen with the lac operon

42
Q

bacteria control expression of …

A

… catabolic operons

43
Q

not all useable carbon sources are …

A

… always available

44
Q

the preferred sugar for metabolism is…

A

…glucose

45
Q

only when glucose is used up are …

A

…other catabolic operons transcribed

46
Q

the lac repressor protein …

A

… represses transcription of the lac operon in the absence of lactose

47
Q

β-galactosidase …

A

…catalyses the hydrolysis of lactose to glucose and galactose

48
Q

β-galactoside permease …

A

… is a transporter protein which allows lactose to enter cell

49
Q

galactoside acetylase is not…

A

…essential

50
Q

What is Lacl?

A

the lac repressor protein

51
Q

What is LacZ?

A

β-galactosidase

52
Q

What is LacY?

A

β-galactoside permease

53
Q

What is LacA?

A

galactoside acetylase

54
Q

in the absence of lactose the repressor (LacI) …

A

…binds to the operator

55
Q

in the absence of lactose the repressor (LacI) binds to the operator

this blocks …

A

…transcription of the operon

56
Q

in the absence of lactose the repressor (LacI) binds to the operator

this blocks transcription of the operon because …

A

…RNA polymerase is unable to move past the repressor

57
Q

in the absence of lactose the repressor (LacI) binds to the operator

this blocks transcription of the operon because RNA polymerase is unable to move past the repressor

because RNA polymerase is blocked…

A

…no transcription of the lacZ, lacY and lacA occurs.

  • therefore there are no catabolic enzymes formed to break down lactose
58
Q

in the absence of lactose the repressor (LacI) binds to the operator

this blocks transcription of the operon because RNA polymerase is unable to move past the repressor

because RNA polymerase is blocked, no transcription of the lacZ, lacY and lacA occurs

therefore there are no catabolic enzymes formed to break down lactose

this is energy efficient as …

A

…they have no substrate to work on

59
Q

when lactose is present it binds to LacI which …

A

…changes its conformation and LacI dissociates from the operator – RNA polymerase is then able to transcribe the lac operon

60
Q

Whats allolactose?

A

an isomer of lactose which acts as the lac inducer

61
Q

this is the DNA sequence to which the Lac repressor (LacI) binds and inhibits transcription of the lac operon

note that the sequence is nearly …

A

…palindromic – this is a feature of many operator sequences

62
Q

binding of LacI to this operator region of the promoter depends on …

A

…those hydrogen bonds between the amino acids of the protein and the bases of the DNA

63
Q

the interaction is …

A

sequence specific – so LacI only binds at this point – it doesn’t stop transcription of other genes or randomly bind to DNA

64
Q

when glucose is present …

A

…transcription is repressed

65
Q

when glucose is present transcription is repressed

this is controlled by a protein called …

A

…CRP (cAMP receptor protein)

66
Q

in the presence of glucose CRP doesn’t bind to …

A

…the CRP binding site

67
Q

in the presence of glucose CRP doesn’t bind to the CRP binding site – because CRP is not bound then RNA polymerase will not …

A

…bind to the promoter

68
Q

CRP requires cAMP to be bound to it in order to …

A

…bind to the CRP binding site

69
Q

cAMP is produced from …

A

…ATP by adenlylate cyclase – glucose inhibits this reaction

70
Q

CRP requires cAMP to be bound to it in order to bind to the CRP binding site

cAMP is produced from ATP by adenlylate cyclase – glucose inhibits this reaction.

this means that when glucose is present in the cell the amount of cAMP is …

A

… low

71
Q

however when glucose molecules are at a low concentration inhibition of cAMP synthesis is lifted and …

A

…cAMP levels start to rise

72
Q

when CRP is bound – as long as lactose is present – then the lac operon is …

A

… transcribed

73
Q

the lac operon has two levels of control:

A

1) repression by LacI in the absence of lactose

2) activation by CRP in the absence of glucose

74
Q

the transcription initiation complex (TIC) itself is a combination of…

A

…many different component proteins that interact with each other

75
Q

binding of the core promoter in eukaryotes is accomplished by …

A

…TATA-binding protein

76
Q

the binding of the initiation complex is itself controlled by interactions with…

A

…transcription factors (activators or repressors) bound to other parts of the promoter

77
Q

genes are regulated at…

A

… many steps

  • transcription of mRNA is only one of these!