Translation Machinery and Mechanism- Lectures 26/27 Flashcards

1
Q

What is tRNA?

A

the link between mRNA and protein

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2
Q

What are the zones of tRNA?

A

DUH arm, anticodon arm (anticodon sequence), V loop, T!C arm, discriminator nucleotide, CCA cap (3’ end), and cognate amino acid

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3
Q

What is the “wobble” position?

A

the third amino acid in a codon that can be variable for some amino acids and does not require the usual complementarity

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4
Q

_____ attaches an amino acid to its cognate tRNA.

A

aminoactyl tRNA synthetase

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5
Q

What are the key things involved in translation?

A

tRNA, ribosomes, eukaryotic translation factors

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6
Q

What composes ribosomes?

A

some protein, but mostly large (40S) and small (60S) subunits that combine to make a 80S which is all composed of rRNA

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7
Q

What are the three eukaryotic translation factors?

A

eIFs
eEFs
eRFs

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8
Q

What are the steps of translation?

A

formation of the pre-initiation complex (43S), initiation, elongation, and termination

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9
Q

What are the three sites on a ribosome?

A

E (exit) site, P (peptidyl) site, A (aminoacytl) site

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10
Q

What composes the pre-initiation complex (43S)?

A

eIF2 (bound to GTP)
Met-tRNA
40S complex (eIF1,3,5)

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11
Q

What is the sequence of events for initiation?

A

eIF4 binds to 5’ cap and recruits 43S complex, which scans down mRNA (using ATP) until the AUG (start codon) is found in the proper context; GTP hydrolysis of eIF2 (via eIF5 on the 43S) results in it dissociating from the complex; all other eIF units leave; 80S ribosome component 60S (guided by eIF5B) binds to UAC and Met-tRNA (at the P site); eIF5B leaves the complex

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12
Q

What are the steps of elongation?

A

next aa-tRNA is escorted to the A site of the ribosome by eEF1A (bound to GTP); hydrolysis of GTP to GDP by GAP on ribosome –> eEF1A leaves complex; peptidyl transferase reaction binds the Met from the Met-tRNA to teh top of the new aa-tRNA; translocation hydrolizes GTP to shift the ribosome so that the Met-tRNA (now cleaved of its Met) is in the E binding site, the new aa-tRNA (now also bound to Met) is in the P binding site, and the A binding site is open for the next aa-tRNA

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13
Q

What regulates which aa-tRNA is used at the P site?

A

the codon-anticodon interaction between the sequence of the mRNA and the anticodon arm of the tRNA which allows for the correct timing for the hydrolysis reaction to proceed

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14
Q

_____ are formed when multiple ribosomes are translating a single mRNA

A

polysomes

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15
Q

What induces termination?

A

a ribosome arriving at a stop codon (this releases it from the mRNA)

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16
Q

Provide examples of common stress signals.

A

Ca2+ release from ER, LPS, heat shock, TNF-alpha, virus infection, GF deprivation, AA starvation, heme deficiency

17
Q

How do cells respond to stress signals?

A

activation of one of a number of protein kinases that phosphorylate eIF2 (initiation factor required to present initiator tRNA to the ribosomal P site)

18
Q

What happens to cells in response to stress (rapamycin) or poor growth conditions?

A

mTOR is inactivated, 4E-BP binds to the eIF4E; overall translation initiation is reduced

19
Q

What happens to cells in the absence of stress (rapamycin) or in good growing conditions or in cancer cells?

A

mTOR phosphorylates 4E-BP; the eIF4 complex is released; overall translation initiation is increased

20
Q

What is SUTR regulation?

A

regulatory proteins that bind to specific UTRs (can be hairpin loop structures formed by intramolecular base pairing) on the mRNA that interfere with scanning

21
Q

_____ is a naturally-occurring mechanism that cells use to down-regulate specific gene expression via inhibitor of small translation.

A

RNAi

22
Q

What can RNAi be used for in research?

A

introduction of exogenous siRNA allows for “knock-down” of expression of a target gene