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Flashcards in RNA Deck (35):
1

Bacterial RNA pol: contains the catalytic site

beta subunit

2

Bacterial RNA pol: Recognizes promoters; required for the initiation of transcription

sigma subunit

3

Proks: Initiates transcription

Holoenzyme (a2BB'sigma)

4

Proks: Carries out transcription

Core enzyme (a2BB')

5

What is also needed for RNA pol.-catalyzed reactions?

Divalent cation

6

Promoters in bacterial genes usually include ~ __ bp upstream of the initiation site and are ___ rich

45 bps, A-T rich

7

Proks: Specific sequence elements in typical promoters (bp location)

-35 Region; Pribnow (-10)

8

What is a closed promoter complex?

When the sigma subunit binds to the core enzyme at a promoter site, however, a conformational change occurs and binding is stabilized

9

What is an open promoter complex

When the two strands of DNA template unwind to produce an unpaired region containing approx. 12 bases.

10

Supercoiling ahead of the transcription bubble

Positive

11

Supercoiling behind the transcription bubble

Negative

12

When does sigma subunit leave the transcription complex?

After about 9 bases are added

13

Proks: Termination

rho independent or rho dependent

14

Termination: DNA templates from which transcription terminates by this mechanism contain an inverted repeat (i.e. 2 palindromic) G-C rich regions followed by 8-10 A's on the template strand (T's on coding). Gives rise to a stem loop structure in the RNA transcript ending in a series of U's.

What type of cells does this occur in?

Rho independent termination; proks

15

Termination: a large hexameric protein binds the 5' end of the new RNA transcript. Protein moves along the RNA strand toward the 3' end using ATP. Polymerase concurrently slows down at a C-rich, G-poor sequence on the DNA template approx. 100 bases from the site of transcription termination. protein catches up to polymerase and uses its helicase function to unwind the RNA transcript from the DNA template strand.

What type of cells does this occur in?

Rho dependent termination; proks

Re: rho protein

16

Rifampicin
-Target
-Function
-What is affected?
-What is not affected?
-What cells are vulnerable?

Target: Beta subunit of bacterial RNA pol
Function: prevents the formation of the first phosphodiester bond
-Affects: Initiation of prok. transcription
-Unaffected: Ongoing transcription
-Vulnerable: Proks

17

Actinomycin D
-Target
-Function
-What is affected?
-What is not affected?
-What cells are vulnerable?

Target: intercalates between DNA strands (major groove)
Function: prevents unwinding of DNA template strands
Affects: initiation of transcription and ongoing transcription
Unaffected: n/a
Vulnerable: proks and euks

18

Euks: what must occur before transcription can occur? Why?

two strands must be unwound; makes promoter region available for appropriate factors to bind

19

Euks: transcribes the precursor large rRNAs (28S, 18S, 5.8S)

What is its location?

RNA Pol I; nucleolous

20

Euks: transcribes the precursor mRNAs (hnRNAs), snRNA, miRNA

What is its location?

RNA Pol II, nucleus

21

Euks: transcribes the precursors of small RNAs (tRNAs, 5s rRNAs)

What is its location?

RNA Pol III, nucleus

22

Euks: promoter regions

CAAT box, GC box, TATA box, Inr (initiator element, pyrimidine rich)

23

Promoter elements that dictate the site of transcription initiation

Basal elements

24

Basal element which directs the binding of RNA pol II

TATA box

25

Binds the TATA box, leading to the assembly of the remaining TFII protein factors

TATA binding protein (TBP)

26

Promoter elements that dictate the rate of transcription initiation

Constitutive elements

27

Constitutive elements (2)

CAAT box and GC box

28

Promoter elements that dictate the specificity of transcription initiation. Mediate transcription in response to environmental signals

Inducible elements

29

Not part of the promoter. regulate transcription by binding regulatory proteins which can either activate or repress transcription

Enhancers

30

Formation of the Pol II pre-initiation complex is followed by: (4)

DNA melting, initiation, promoter clearance, and transcription elongation.

31

During initiation, RNA pol II does not bind DNA directly but binds to:

proteins (first TBP then TFs then RNA poly) which are bound to DNA

32

Euks: Necessary but not sufficient for effective transcription initiation

Basal apparatus

33

Binding of the pol. is stabilized by a conromational change in the DNA caused by the binding of regulatory proteins to ___________(2) as well as ___________ (protein type) bound to enhancers

constitutive and inducible promoter elements; activator proteins

34

Alpha-amanitin
-Target
-Function
-What is affected?
-What is not affected?
-What cells are vulnerable?

Target: RNA pol II (lesser extent for RNA pol III)
Function: inactivates RNA pol II
Affected: mRNA synthesis inhibited, liver failure and death (48 hours) - essential enzymes degraded and not replaced
Not affected: RNA pol I
Vulnerable: euks

35

Euks: Termination

AAUAAA polyadenylation sequence