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Molecular Test 3 > Lecture 19 > Flashcards

Lecture 19 Flashcards

1
Q

How do we look at transcription complexes?

A

Take native chromatin from the nuclei.

RNA pol III only transcribes genes that are programmed into ACTIVE transcription complexes.

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2
Q

What transcription factors provide the signal that the gene is ready to be transcribed?

A

TFIII A, TF III B, and TFIIIC

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3
Q

What happens once TFIIIA-C bind to the ICR?

A

RNA pol III can bind and transcribe

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4
Q

How did the scientists see what TFs were needed? 3 steps.

A
  1. Did in invitro assay and purfied chromatin.
  2. Added DNA pol III (purified)
  3. Look for expression
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5
Q

What did the polymerase assay using chromatin tell?

A

Genes that were normally off were activated, which is why fingers 4,5, and 6 give more expression.
More active transcription complexes are formed on OOCYTE type 5S RNA genes.

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6
Q

What finger mutant displays the highest level of gene activity?

A

H183N

finger 6

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7
Q

Explain the feedback inhibition of Wild Type TFIIIA (3 steps)

A
  1. TFIIIA-C bind to the ICR of the 5S Gene
  2. 5S RNA with an affinitiy for TFIIIA is transcribed.
  3. Free TFIIIA binds to the 5S RNA forming the 7S particle
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8
Q

What did the titration experiment with the finger 6 mutant show ?

A

When the 5S RNA is transcribed from the mutant TFIIIA, the free TFIIIA mutatants won’t bind the 5SRNA as well. The TFIIIA stays high so more complexes can be transcribed.

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9
Q

Two ribosomal protiens

A

60S (L1 to L49)

40S (S1 to S33)

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10
Q

How do cells make the right amount of ribosomal protein compared with mRNA synthesis?

A

5S RNA can bind to TFIIIA OR Protein L5

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11
Q

Protein L5

A

can displace TFIIIA from the 5S mRNA!!

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12
Q

Explain how L5 stimulates transcription of 5S RNA

A

L5 displaces the TFIIIA off the 7S particle.
L5 on the 5S RNA is incorporated into the ribosome, and the free TFIIIA can bind to more transcription complexes to make more 5S RNA.

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13
Q

What is the difference between RNA pol III and RNA pol II

A

RNA pol III is housekeeping

RNA pol II tells TISSUE SPECIFICITY

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14
Q

Studies of promoter analysis:

A

Tell WHERE and WHEN

What controls SPATIAL and TEMPORAL activity

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15
Q

Studies of basic mechanisms:

A

how it actually works
how it gets signal
HOW AND WHY

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16
Q

UAS

A

upstream activating sequence
can be far away
CIS acting

17
Q

Core promoter

A

shortest sequence of DNA at which RNA pol can initiate transcription and assemble basal transcription apparatus.

18
Q

What does the core promoter include?

A

Inr, TATA, and DPE

19
Q

Enhancers

A

increase utilization of the promoter
can be upstream or downstream
CIS acting

20
Q

Response elements

A

bind inducible factors to respond to different conditions

21
Q

Silencer

A

sequence of DNA that can inactivate expression of a gene in its vicinity

22
Q

Promoter

A

stretch of DNA to provide temporal/spatial/rate regulation of the gene
CIS acting

23
Q

Deletion Mapping

A

exonuclease gets rid of parts of promoters and tells what is important by observing changing transcription

24
Q

Linker Scanning

A

maintains spacing, so insertions may not have as much effect when in between enhancer and promoter

25
Q

DNA footprinting

A

Shows were different things bind since DNAase can’t cut them

26
Q

3 DNA binding domains

A

Zinc fingers
Homeodomains
POU domain

27
Q

Homeodomain

A

Tx factor turns on gene in the wrong location

Alpha helix-turn-helix

28
Q

Helix 1

A

10-22

29
Q

Helix 2

A

28-37

30
Q

Helix 3

A

42-58

binds in the major groove

31
Q

POU domain P

A

pituitary

32
Q

POU domain O

A

oct tx factors

33
Q

POU domain U

A

unc

uncoordinated tx factors

34
Q

POU domain

A

helix turn helix

3 types of tx factors that share same DNA binding domain

Molecular Test 3 (9 decks)

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