Lecture 16 Flashcards

1
Q

Gene expression in prokaryotes compared to eukaryotes:

A
  • Ribosomes in eukaryotes are bigger
  • Transcription and translation are not coupled
  • Internal translation initiation within an mRNA does not usually occur in eukaryotes
  • Monocistronic mRNA
  • Ribosomes are synthesised in the nucleolus and transported to the cytoplasm in eukaryotes
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2
Q

Translation in prokaryotes:

A

Mechanics between prokaryotes and eukaryotes are very similar

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3
Q

Cap recognition:

A
  • Important in translation
  • Indicates where the 5’ end of the transcript is
  • Important in determining where initiation is going to happen
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4
Q

Pre-initiation compex:

A
  • Scans along the RNA until a ATG is reached
  • Base pairing between the ATG and tRNA occurs
  • 60S subunit binds to elongation to occur
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5
Q

Translation initiation codon:

A
  • Most translational initiation occurs at the first AUG, closest to the 5’ cap
  • Efficiency is influenced by fit to a consensus sequence
  • Usually (gcc)gccRccAUGG sequence
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6
Q

Dicistronic mRNAs:

A
  • Internal translation initiation via IRES (internal ribosome entry sites)
  • Recognition of 4F
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7
Q

mRNA translatability:

A
  • 5’ cap and polyA tail
  • PolyA tail determines if the mRNA will be translatable
  • De-capping or shortening polyA tail precipitates degradation of mRNA
  • A series of polyA binding proteins regulate translation initiation, recruit enzymes to add A’s,
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8
Q

What does polyA tail length determine?

A
  • It will determine how well it is translated
  • The longer the tail, the more translation occurs
  • A mechanism of regulating when you will get the product
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9
Q

Nonsense mediated mRNA decay (NMD):

A
  • An RNA surveillance mechanism
  • Directs degradation of mRNAs containing premature stop codons
  • Prevents production of truncated gene products and defected proteins
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10
Q

NMD mechanism:

A
  • Ribosomes displace exon junction complexes on mRNAs
  • When premature termination due to nonsense mutation occurs this is due to ribosomes neglecting to displace complexes.
  • Upf decaps transcripts and nucleases can degrade the entire mRNA
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11
Q

Premature termination due to abnormal splicing produces:

A
  • UpF triggers mRNA degradation

- The mRNA is degraded

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12
Q

Non-stop mediated decay means that splicing is fine, but there is an error changing the reading frame so the correct stop codon is missed:

A
  • Ribosome translates polyA tail (lys) and stalls at the end
  • The poly-lycine tail produces a localised depletion of lycine, so the ribosome stalls to wait for more activated tRNAs.
  • The stalling causes the nonstop mediate decay complex to interact, kick off ribosome and an exosome will degrade from the 3’ end.
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13
Q

Programmed frame-shifting:

A
  • Gag protein is generated when no frameshift mutation is present
  • When a frameshift mutation is present everything in the protein is different from the frameshift onward
  • A number of genes are overlapping in the viral genome, so frameshifting is not a problem! The frameshift produces a different protein
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14
Q

Polypeptide maturation is a post translation method of modifying:

A
  • Folding, co-factor binding, interaction with other polypeptides
  • Chaperones assist correct folding, using energy from ATP hydrolysis
  • Incorrectly folded and aberrant proteins are degraded
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15
Q

Ubiquitin:

A
  • Degraded by covalent attachment of Ub to lysine
  • Extent and position of ubiquitiation affects the fate of the protein
  • Attachment of Ubiquitin doesn’t always mean the protein will be degraded
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16
Q

Mono ubiquitylation:

A
  • Histone regulation
17
Q

Multi-ubiquitylation:

A

Endocyctosis

18
Q

Poly-ubiquitination

A
  • Ub joined end to end on the protein
  • Proteasomal degradation
  • DNA repair
19
Q

Inteins:

A
  • Intervening protein segments
  • Removed from proteins by self-splicing (autocatalytic)
  • Rejoin flanking protein (exteins)
  • Similar to transposons but are protein based, not DNA based.