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Lecture 7
Transcription (mRNA Synthesis) and Translation (Protein Synthesis)

BIOSCI 101: Cellular and Molecular Biology

Transcription and Translation

Transcription is the process of transfer of information from DNA into RNA (reverse transcriptase achieves the reverse process) The existence of a messenger molecule which transferred genetic information from the nucleus to the cytoplasm was first hypothesised by Jacob and Monod Information transfer is completed by translating the nucleotide sequence of mRNA into the amino acid sequence of a protein
BIOSCI 101: Cellular and Molecular Biology

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Information flow from gene to protein

BIOSCI 101: Cellular and Molecular Biology

Transcription and Translation (contd)

Synthesis of messenger RNA (transcription) a) an enzyme called RNA polymerase binds to a nucleotide sequence known as the promoter - as RNA polymerase moves along the DNA molecule the two strands of the molecule separate b) the polymerase assembles ribonucleoside triphosphates into RNA one at a time - same base-pairing principle that governs DNA replication (rA pairs with dT, rG with dC, rC with dG, rU with dA)
BIOSCI 101: Cellular and Molecular Biology

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Stages of transcription

Campbell 17.7

BIOSCI 101: Cellular and Molecular Biology

Elongation a closer look

Campbell 17.9

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Transcription (contd) c) synthesis is in a 53 direction as the enzyme reads the template DNA strand in a 35 direction - as the enzyme moves along, the DNA helix rewinds, displacing the newly made mRNA molecule - transcription ceases when the polymerase reaches a termination signal and releases the DNA template and mRNA product d) the finished mRNA strand is antiparallel and complementary to the template strand from which it was transcribed
BIOSCI 101: Cellular and Molecular Biology

Transcription and Translation (contd)

Transcription (contd) e) both strands may code for mRNA and some genes may overlap f) the average mRNA molecule is 900 - 1200 bases long (sufficient to code for an average polypeptide 300 - 400 amino acids long) - in prokaryotes mRNAs may code for more than one polypeptide

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Transcription (contd) g) in eukaryotes mRNA is extensively modified after transcription - enzymes add a 5 cap and a 3 polyA tail to the mRNA - the mRNA molecule is edited by a process known as splicing - intervening sequences (introns) are cut out and expressed sequences (exons) are ligated together to form the mature mRNA molecule
BIOSCI 101: Cellular and Molecular Biology

RNA processing: addition of the 5 cap and polyA tail

Campbell 17.10

BIOSCI 101: Cellular and Molecular Biology

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RNA splicing (1)

Campbell 17.11

BIOSCI 101: Cellular and Molecular Biology

RNA splicing (2)

Campbell 17.12

BIOSCI 101: Cellular and Molecular Biology

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Exons may encode protein domains

Campbell 17.13

BIOSCI 101: Cellular and Molecular Biology

Translation the basic concept

Campbell 17.14

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Protein Synthesis (Translation) a) mRNA - the sequence of bases in DNA and then mRNA specifies the amino acid sequence - AUG = start codon, UAA, UAG and UGA = stop codons (see next lecture) - every mRNA molecule contains a ribosomal binding site at its 5 end

BIOSCI 101: Cellular and Molecular Biology

Transcription and Translation (contd)

Translation (contd) b) ribosomes - ribosomes bind to the 5 end of newly transcribed mRNA - the small ribosomal subunit has a binding site for the mRNA molecule - the large subunit has three binding sites for tRNA molecules ( (the p peptidyl p y site, , the aminoacyl site and the exit site)

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Translation (contd) - the peptidyl (P) site holds the tRNA carrying the growing polypeptide chain - the aminoacyl (A) site holds the tRNA carrying the next amino acid to be added - the exit (E) site is the site from which discharged tRNAs leave the ribosome

BIOSCI 101: Cellular and Molecular Biology

tRNA molecules

Growing polypeptide

Exit tunnel

Large subunit

E PA
Small subunit 5 mRNA 3

Anatomy of a ribosome

P site

Exit tunnel A site

E site

E P A
mRNA binding site

Large subunit Small subunit

Campbell 17.17

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Transcription and Translation (contd)

Translation (contd) c) ) tRNA - adapter molecules which bring amino acids into register on the message - for each of the 20 amino acids there is at least one specific tRNA molecule present in the cell - the tRNA molecule possesses a site for attachment of the amino acid and also an anticodon loop to pair with the mRNA codon - specific enzymes (aminoacyl tRNA synthetases) recognise the tRNAs and attach the correct amino acid
BIOSCI 101: Cellular and Molecular Biology

The structure of transfer RNA

Campbell 17.15

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Specific amino acids are joined to tRNA

Campbell 17.16
BIOSCI 101: Cellular and Molecular Biology

Transcription and Translation (contd)

Translation (contd) d) Synthesis of a polypeptide involves three stages:i) initiation - the codon AUG specifies methionine and is the most commonly used start signal - the smaller ribosomal subunit attaches to the 5 end of the mRNA - the first tRNA molecule bearing the amino acid met, plugs into the initiator codon AUG on the mRNA molecule - the larger ribosomal subunit locks into place with the met tRNA occupying the P site - the A site is vacant and the initator complex is now complete
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Translation 1:Initiation

Campbell 17.18

BIOSCI 101: Cellular and Molecular Biology

Transcription and Translation (contd)

Translation (contd) ii) elongation - a second aminoacyl tRNA binds to the codon in the A site - a component of the larger subunit catalyses the formation of a peptide bond between the two amino acids brought together at the ribosome At the same time ribosome. time, the bond between the first amino acid and its tRNA is broken

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

elongation (contd) - the ribosome translocates ie moves along the mRNA chain in a 5 to 3 direction and the second tRNA, with the mRNA and the dipeptide attached, is moved from A to P - the first tRNA moves from P to E and is released from the ribosome - a third tRNA moves into the A site and peptide p bond is formed. This step p is another p repeated until the polypeptide is complete

BIOSCI 101: Cellular and Molecular Biology

Translation 2:Elongation

Campbell 17.19

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Translation (contd) iii) termination - the ribosome reaches a termination codon (UAA, UAG or UGA) - release factor binds to the A site - the polypeptide is cleaved from the last tRNA which is then released - the th mRNA RNA and d ribosomal ib l subunits b it dissociate di i t

BIOSCI 101: Cellular and Molecular Biology

Translation 3: Termination

Campbell 17.20

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Translation (contd) - more than one ribosome binds to each mRNA - a polyribosome consists of a single mRNA along which are travelling a series of ribosomes to which in turn are attached growing peptide chains

BIOSCI 101: Cellular and Molecular Biology

Polyribosomes

Campbell 17.21

BIOSCI 101: Cellular and Molecular Biology

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Transcription and Translation (contd)

Translation (contd) e) Modification of the product - many proteins are modified in the ER or the Golgi after they are made - many hormone molecules mature via a cleavage reaction - glycosylation is the addition of carbohydrate residues (may occur even before the peptide is completed) - signal sequences at the N-terminus of the polypeptide target proteins destined for secretion to the ER and Golgi
BIOSCI 101: Cellular and Molecular Biology

Proteins may be targeted to the ER

Campbell 17.22

BIOSCI 101: Cellular and Molecular Biology

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Test question
Which of the following is NOT directly involved in the process of translation? 1. mRNA 2. rRNA 3. tRNA 4. aminoacyl-tRNA synthetase 5. ligase

BIOSCI 101: Cellular and Molecular Biology

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