Gene Structure 4 Flashcards

1
Q

Who came up with the intron early theory?

A

Walter Gilbert

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2
Q

What is the intron early theory?

A

That introns originated in prokaryotes but were lost due to genome streamlining - suggesting that eukaryotes inherited all introns from prokaryotic ancestors.
Believed introns promoted gene evolution as they permitted the shuffling of genes - produced more complex genes and larger protein collection.

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3
Q

What is the intron late theory?

A

Stated that introns were unique to eukaryotes and that new introns have been emerging throughout eukaryotic evolution - prokaryotes have never contained introns or the spliceosome.

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4
Q

What is the current intron theory?

A

Somewhere in between the early and late theory -
Group II introns originated in the mitochondrial ancestor and these invaded the host genome upon mitochondrial endosymbiosis.
(still debated).

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5
Q

Why can introns be a burden to a host?

3

A

Cell must contain spliceosome and this is a huge complex.
Also increased energy cost as RNAPII is transcribing larger sequences.
Create vulnerability - as need recognition by cis regulatory sequences to be spliced.

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6
Q

What are the 5 “life phases” of introns?

A
Genomic intron. 
Transcribed intron. 
Intron being spliced. 
Excised intron. 
Exon junction complex (EJC)-harbouring transcript.
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7
Q

What is the genomic intron?

A

Still in the DNA and is the location of the gene’s cis regulatory elements.

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8
Q

How are genomic introns involved in transcription initiation?

A

They contain enhancers, silencers and TF binding sites - mostly found in 5’ introns.

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9
Q

How are genomic introns involved in alternative splicing?

A

They contain alternative transcriptional initiation sites (promoters).
e.g. alpha-fetoprotein - cell specific expression.

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10
Q

How are genomic introns involved in transcriptional termination?

A

Intron sequences can regulate polyadenylation and cleavage.

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11
Q

Give an example of how genomic introns effect transcriptional termination.

A

Human Beta globin - protein can be soluble or membrane bound depending on where transcription is terminated and poly A tail is added.
Soluble - polyA on exon 14.
Membrane bound on exon 13.

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12
Q

What can genomic introns also be host to?

A

Nested genes - protein coding or non-coding RNA

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13
Q

What is the genomic design hypothesis?

A

Genes that need complex regulation contain more introns to accommodate their regulatory elements.
Highly expressed genes, which need low regulation have shorter introns.

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14
Q

What is a transcribed intron?

A

An intron that is present in the pre-mRNA

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15
Q

What is the RNAPII elongation rate and what does this mean for intron transcription?

A

50kb min^-1.
This means that intron may take hours to be transcribed - this explains the time delay between gene activation and translation of protein.

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16
Q

Give an example of how the gene activation to translation of protein time delay involved in negative feedback loops

A

Example - HES7 gene (transcription factor) - 19 minute delay between transcriptional initiation and full mature mRNA.
The protein produced inhibits its own transcription.

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17
Q

What is a spliced intron?

A

An intron present in mRNA - being spliced out by the spliceosome.

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18
Q

Which process is intron splicing linked to and how?

A

Transcription - linked via C-terminal domain of RNAPII.

19
Q

How can splicing effect initiation of transcription?

A

U1-snRNP associates with and recruits TFIIH and TFIID to the 5’ splice site and stimulates phosphodiester bond formation (in mRNA).

20
Q

How can splicing effect elongation of transcription?

A

Splicing factors and spliceosomal components can interact with transcriptional elongation factors.
U2 snRNP binding at the 3’ end of the transcript promotes the elongation using RNAP.

21
Q

How can splicing effect termination of transcription?

A

U2 snRNP interacts with cleavage/polyA specificity factor (CSPF).
CSPF binds to polyA site and this can promote polyA tail formation and/or splicing via interaction with U2.

22
Q

What is an excised intron?

A

An intron that has been removed from pre-mRNA by the spliceosome.

23
Q

What often happens to excised intron?

A

They undergo debranching and degradation.

24
Q

What can be expressed after an intron has been excised?

A

Embedded/nested genes - often miRNA and snoRNAs.

25
Q

How long are miRNAs?

A

~22nt

26
Q

Which organisms are miRNA found in?

A

Metazoans, plants and other eukaryotes.

27
Q

How are miRNAs expressed if embedded in introns?

A

They are co-expressed with other genes because the lack promoters.

28
Q

Where do miRNAs bind to and what does this result in?

A

Bind to 3’UTRs of mRNA and cause degradation/prevent translation.

29
Q

How long are snoRNAs?

A

60-150nt

30
Q

Which organisms are snoRNA found in?

A

Archaea and eukaryotes.

31
Q

What do snoRNAs do?

A

Modify RNAs - rRNAs, snRNAs, tRNAs.

Can also regulate mRNA and are released after splicing.

32
Q

What are the two classes of snoRNAs?

A

C/D box snoRNAs - guide 2’O methylation of rRNAs.

H/ACA box snoRNAs - guide pseudoridylation of rRNAs.

33
Q

What is another feature of snoRNAs?

A

They are mobile genetic elements via retro-transposition.

34
Q

Where does the EJC bind?

A

Binds ~25nt upstream of the exon-exon junction (where an intron has been) on the mRNA transcript.

35
Q

What are the four core proteins in the EJC?

A

MAGO, Y14, eF4AIII and MLN51.

36
Q

What are the four roles of the EJC?

A

Nuclear transport
Translation activation
mRNA localisation
nonsense mediated decay.

37
Q

How is the EJC involved in nuclear transport?

A

EJC (that is bound to the mRNA) interacts with nuclear pore complex and recruits the export factor - ALY/REF.

38
Q

How much faster is the nuclear transport export rate for spliced transcripts?

A

10x faster.

39
Q

How is EJC involved in translation activation?

A

MLN51 interacts with eIF3, which is a translational initiation factor.

40
Q

How is EJC involved in cytoplasmic localisation?

A

Some mRNAs require the EJC to localise them to subcellular locations.

41
Q

What is nonsense mediated decay?

A

Degradation of mRNAs that include premature stop codons.

42
Q

Why do cells have a mechanism in place to degrade mRNAs that have premature stop codons?

A

To prevent dominant-negative/gain of function.

43
Q

How is EJC involved in nonsense mediated decay?

mechanism

A

UPF3 loaded on to EJC during splicing.
UPF2 binds complex in the cytoplasm – ribosome knocks off EJCs unless there is a premature stop codon.
SURF complex forms and binds to ribosome at the premature stop codon.
Because the ribosome has stopped the EJC is left on the mRNA – UPF1 then interacts with UPF2.
UPF1 is phosphorylated, allowing it to bind the RNA and recruit SMG6 and SMG5-SMG7
SMG5-7 promoter deadenylation and decapping.
SM6 cleaves the mRNA.