lecture 4 Flashcards

1
Q

what are the steps in transcription?

A
  • initiation
  • elongation
  • termination
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2
Q

what supports transcription?

A
  • transcription regulators
  • transcription factors
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3
Q

what is transcription?

A

copy the DNA into RNA

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4
Q

how many RNA polymerases do eukaryotes have?

A

3

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5
Q

how many RNA polymerases fo bacteria have?

A

1

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6
Q

what is the structure of bacteria RNA polymerase?

A

5 subunits
- 400-450 kDa

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7
Q

what is the function of RNA polymerases?

A

reading and transcribing a gene sequence into an RNA product

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8
Q

is RNApol a conserved structure?

A

RNA polymerase general architacture and catalytic function is conserved across the three domains of life

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9
Q

what is the action of bacterial RNA pol?

A
  • bacterial RNAP has a crab-cpaw appearance that opens and closes during transcription
  • the incoming dsDNA goes into the cleft, becomes unwound and is transcribed into a strand of RNA
  • the exiting DNA comes out in a perpendicular direction
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10
Q

what is the structure of the core of RNA polymerase?

A

parts of the beta and beta’ and the two alpha subunits

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11
Q

what is the structure of the jaw-lobe of RNA polymerase?

A

parts of the beta subunit

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12
Q

what is the role of sigma in bacterial RNA polymerase?

A

participates in recognizing the promoter and initiate transcription

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13
Q

where does the sigma factor bind on bacterial RNA polymerase?

A

to the cleft

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14
Q

what happens in the initiation stage of transcription?

A

RNAP binds to the promoter, near the beginning of a gene transcription bubble

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15
Q

what happens in the elongation stage of transcription?

A

RNAP reads the template strand one base at a time
RNAP buikds a complementary RNA molecule

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16
Q

what happens in the termination stage of transcription?

A

Terminators signal that the RNA transcript is complete RNAP is released

17
Q

what is the bacterial promoter region in a gene?

A

specific dsDNA region where the RNAPol binds

promoter regions have very conserved structure

18
Q

what are the regions of the promoter of a bacterial gene?

A

UP element: -37 to -58
the -35 element: -35 to -30
the extended -10 element (Ext): -17 to -14
the -10 element: -12 to -7
the discriminator element (Dis): -6 to -4

*the exact position of each element may vary

19
Q

how does RNAPol recognize the bacterial gene promoter?

A

specific elements are recognized by individual domains of sigma factor and alpha-RNAPol

20
Q

where does transcription start in the gene?

A

at the +1 bp

21
Q

what is the composition of E.coli during growing conditions?

A

55% protein
10% lipid
3% DNA
20% RNA
<1% mRNA

22
Q

why is RNA found in much higher quantities within the bacterial cell than mRNA?

A

mRNA has a half-life od 3-8mins
RNA has a half-life of days

23
Q

what is E.coli house keeping sigma factor?

A

sigma 70

24
Q

what is the role of sigma70 (house keeping sigma factor)?

A

has lots of domains, each responsible for recognising a section of promoter
- sigma 70 recognizes most promoters

25
Q

what are the domains of sigma70?

A

3&4 - correct position of the promoter
1&2 - open complex

26
Q

what mediates binding of RNAP to the promoter?

A

initiation factor

27
Q

what is the role of sigma factor in initiation of transciption?

A

ensures the correct orientation of the dsDNA in the RNApol

28
Q

what does elongation of the RNA transcript occur?

A

once the sigma factor is released

29
Q

what is the role of RNAPs catalytic core?

A

RNAP catalytic core is capable of RNA polymerization

30
Q

what eukarotic polymerase is similar to bacterial RNA pol?

A

Pol II

31
Q

what is the structures involved in elongation?

A

Initiation factors are exchanged for elongation factors during the early stages of RNA synthesis

in E.coli- initiation factor (sigma 70) and elongation factor (NusG) bind to the same RNAP region

32
Q

what happens during elongation?

A

DNA-RNA hybrid of 8 to 9 base pairs is formed in the transcription bubble
The downstream DNA is not yet transcribed; the upstream DNA has regained the duplex form

33
Q

what is the function of backtracking in elongation during transcription?

A

incorrectly incorporated nucleotides can be excised

34
Q

what is the role of the rudder loop in elongation?

A

separates DNA

35
Q

what maintains the transcription bubble in elongation?

A

a series of loops

36
Q

where is the active site of the RNApolymerase?

A

the active site is located at the floor of the cleft

37
Q

when does the elongation phase start?

A

after RNAP synthesizes 9-10 RNA nts, it escapes from the promoter region

38
Q

what occurs during the termination of bacterial transcription?

A

intrinsic termination- DNA and RNA sequence
the helicase Rho associate with C-rich RNA sequences and expeles RNAP from the DNA

the helicase Rho engages with NusG, NusA, and RNAP during transcription elongation

RNAP dissociation from the DNA