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Lecture 8

The Genetic Code and p Control Transcriptional

BIOSCI 101: Cellular and Molecular Biology

Genetic code/transcriptional control

Information Transfer in Pro Pro- and Eukaryotes - in prokaryotes, mRNA may be polycistronic - transcription and translation can be occurring simultaneously in prokaryotes - mRNA molecules undergo post-transcriptional processing in eukaryotes - in eukaryotes transcription and translation are separated by the nuclear membrane
BIOSCI 101: Cellular and Molecular Biology

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The flow of genetic information

Campbell 17.3

BIOSCI 101: Cellular and Molecular Biology

Coupled transcription and translation in bacteria

Campbell 17.25

BIOSCI 101: Cellular and Molecular Biology

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Transcription and translation in eukaryotes k t

Campbell 17.26

BIOSCI 101: Cellular and Molecular Biology

Pro- and eukaryotic mRNA

BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code a) the code is a triplet of nucleotides - a triplet of nucleotides is called a codon - for a triplet code, the possible combinations are 4 x 4 x4 = 64

BIOSCI 101: Cellular and Molecular Biology

The triplet code

Campbell 17.4

BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code (contd) b) The code was deciphered using in vitro systems - a cell-free (in vitro) system for protein synthesis consists of extracts of E. coli containing ribosomes, tRNA molecules and bacterial mRNA - radioactive amino acids are incorporated into an acid insoluble product acid-insoluble - experiments using synthetic mRNA molecules demonstrated that i) mRNA is read sequentially ii) how it is read depends on the reading frame
BIOSCI 101: Cellular and Molecular Biology

Deciphering the genetic code

BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code (contd) c) The code is degenerate - most amino acids are specified by more than one triplet - there is >1 tRNA for some amino acids eg leucine - some tRNA molecules require accurate basepairing only at the first two positions of the codon and can tolerate a mis-match at the third (the wobble hypothesis) eg arginine
BIOSCI 101: Cellular and Molecular Biology

The genetic code

Campbell 17.5
BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code (contd) d) The code is more or less universal - the most usual start codon is AUG and the stop codons are UAA, UAG and UGA - comparison of nucleic acid and protein sequences of molecules from a variety of proand eukaryotes confirmed the accuracy and universality of the code - some differences in mitochondrial DNA coding
BIOSCI 101: Cellular and Molecular Biology

The genetic code is universal

Campbell 17.6

BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code (contd) e) Changes in sequence cause mutations - a mutation is any permanent, inheritable alteration in the DNA sequence - may arise spontaneously, as a result of low frequency random errors, or may be induced by external agents which damage DNA

BIOSCI 101: Cellular and Molecular Biology

Genetic code/transcriptional control (contd)

The genetic code (contd) - point mutations involve a change in a single base pair. Can arise from : i) mistakes in replication by DNA polymerase ii) mutagenic agents eg UV light iii) spontaneous chemical reactions in cells

BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code (contd) f) the consequences of a point mutation i) silent - no effect on the amino acid sequence ii) missense - an amino acid is changed, may not be serious iii) nonsense - creation of a termination codon, usually serious iv) frameshift - the reading frame is altered by insertion or deletion of a base. Changes many amino acids and usually causes complete loss of function

BIOSCI 101: Cellular and Molecular Biology

Changes may not be detected

Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3

mRNA5 A U G A A G U U U G G C U A A 3 Protein Met Lys Phe Gly Stop Amino end Carboxyl end

(a) Nucleotide-pair substitution: silent

A instead of G
3 T A C T T C A A A C C A A T T 5 5 A T G A A G T T T G G T T A A 3

U instead of C
5 A U G A A G U U U G G U U A A 3

Met Campbell 17.24

Lys

Phe

Gly

Stop

BIOSCI 101: Cellular and Molecular Biology

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Nucleotide-pair substitutions

Campbell 17.24
BIOSCI 101: Cellular and Molecular Biology

A single base change causes sickle-cell disease

Wild-type yp hemoglobin g 3 5 Wild-type hemoglobin DNA C T T 5 G A A 3 Sickle-cell hemoglobin g 3 5 Mutant hemoglobin DNA C A T G T A 5 3

mRNA 5 G A A 3 5

mRNA G U A 3

Normal hemoglobin Glu

Campbell 17.23

Sickle-cell hemoglobin Val

BIOSCI 101: Cellular and Molecular Biology

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Campbell 5.21

BIOSCI 101: Cellular and Molecular Biology

Normal vs mutant haemoglobin

Campbell 5.21

BIOSCI 101: Cellular and Molecular Biology

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Mapping malaria and the sicklecell allele

Campbell 23.17

BIOSCI 101: Cellular and Molecular Biology

Nucleotide-pair insertions or deletions

Campbell 17.24
BIOSCI 101: Cellular and Molecular Biology

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Genetic code/transcriptional control (contd)

The genetic code (contd) g) cells protect themselves from mutations - fewer than 1 / 1000 random changes causes a mutation - most mutations are eliminated by DNA repair mechanisms

BIOSCI 101: Cellular and Molecular Biology

Excision repair i of f DNA damage

Campbell 16.19
BIOSCI 101: Cellular and Molecular Biology

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A practice question
There are 61 mRNA codons that specify an amino acid, but only about 32 tRNAs are actually required to translate them. This is best explained by the fact that: A) some tRNAs have anticodons that recognize four or more different codons. B) the rules for base pairing between the third base of a codon and tRNA are flexible. C) many codons are never used, so the tRNAs that recognize them are dispensable. D) Some tRNAs are easily destroyed by nucleases.

BIOSCI 101: Cellular and Molecular Biology

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