1) The genetic code uses triplets of nucleotides (codons) to specify the 20 amino acids. Each codon is made up of 3 nucleotides (A, U, G or C).
2) One codon specifies one amino acid, so there are 64 possible codons (4x4x4) to code for the 20 amino acids. Many codons code for the same amino acid, making the code degenerate.
3) Early experiments by Nirenberg and colleagues using cell-free systems established that the genetic code is read in triplets, is universal across organisms, and that the probability of codon usage depends on the relative concentrations of nucleotides.
1) The genetic code uses triplets of nucleotides (codons) to specify the 20 amino acids. Each codon is made up of 3 nucleotides (A, U, G or C).
2) One codon specifies one amino acid, so there are 64 possible codons (4x4x4) to code for the 20 amino acids. Many codons code for the same amino acid, making the code degenerate.
3) Early experiments by Nirenberg and colleagues using cell-free systems established that the genetic code is read in triplets, is universal across organisms, and that the probability of codon usage depends on the relative concentrations of nucleotides.
1) The genetic code uses triplets of nucleotides (codons) to specify the 20 amino acids. Each codon is made up of 3 nucleotides (A, U, G or C).
2) One codon specifies one amino acid, so there are 64 possible codons (4x4x4) to code for the 20 amino acids. Many codons code for the same amino acid, making the code degenerate.
3) Early experiments by Nirenberg and colleagues using cell-free systems established that the genetic code is read in triplets, is universal across organisms, and that the probability of codon usage depends on the relative concentrations of nucleotides.
1) The genetic code uses triplets of nucleotides (codons) to specify the 20 amino acids. Each codon is made up of 3 nucleotides (A, U, G or C).
2) One codon specifies one amino acid, so there are 64 possible codons (4x4x4) to code for the 20 amino acids. Many codons code for the same amino acid, making the code degenerate.
3) Early experiments by Nirenberg and colleagues using cell-free systems established that the genetic code is read in triplets, is universal across organisms, and that the probability of codon usage depends on the relative concentrations of nucleotides.
Code CHARACTERISTICS OF THE GENETIC CODE: 1. In linear form Uses the 4 ribonucleotides on mRNA (A, U, G, C) Sequence is complimentary to nucleotide sequence of DNA 2. In triplet Codon language of mRNA Each word is made of 3 nucleotides specifying 1 amino acid
mRNA and Triplet Code Words
4. Degenerate a given amino acid can be specified by more than one triplet codon GUU
GUA
GUC
GUG
Valine Valine is an example of the degeneracy of the code. All four of these triplets code for valine. 5. Contains start signals- initiate translation stop signals- terminate translation AUG
UAA
Start (Methionine)
UAG
UGA
Stop
6. Commaless once translaton begins, codons are read continuously without breaks no internal punctuation
7. Nonoverlapping each nucleotide is part of only 1 codon 8. Universal (nearly) With minor exceptions, all organisms use a single coding dictionary Early Studies that Established the Basic Operational Patterns of the Genetic Code In the late 1950s, researchers thought that DNA itself might directly encode proteins during their synthesis. Francois Jacob and Jacques Monod suggested that an RNA molecule was intermediate between DNA and proteins
Once this mRNA was discovered, the
question was how could just 4 bases code for 20 amino acids. Sidney Brenner Argued that the code must be a triplet since three letter words represent the minimal use of four letters to specify 20 amino acids. A code of 4 nucleotides taken 2 at a time 42 = 16 nucleotides A code of 4 nucleotides taken 3 at a time 43 = 64 nucleotides Francis Crick and colleagues Frameshift Mutation ! Occurs as a result of the addition or deletion of one or more nucleotides in the gene. ! Gain or loss of one or two nucleotides will shift the frame of reading. ! When three nucleotides are involved, the frame of reading is reestablished. ! This would not occur if the code is not triplet.
! amino acids were labeled
! enzyme to be used was isolated bacteria and it functions to degrade RNA " discovered in 1955 by Marianne Grunberg-Manago and Severo Ochoa " in vitro, with high concentrations of ribonucleosides dephosphates, the reaction can be forced to synthesize RNA " unlike, RNA polymerase, it does not require a DNA template Use of Homopolymers Steps: 1. In a series of experiments, RNA polymers were synthesized using just one type of ribonucleotide. 2. Researchers labeled 1 of 20 amino acids added. 3. Each RNA were tested to determine which, if any, amino acids were incorporated into the newly synthesized proteins. Result:
Deciphering the Code
Cell- Free Polypeptide Synthesis ! Because the enzyme used in this experiment does not require a template, each addition of a ribonucleotide is random, based on the relative concentration of the ribonucleoside diphosphates. ! The probability of the insertion of a specific ribonucleotide is proportional to the availability of that molecule, relative to other available ribonucleotides. ! must contain the essential factors for protein synthesis: ribosomes, tRNA, amino acids, and other molecules essential to translation
Mixed Copolymers !
Makes use of RNA heteropolymers- 2 or
more different ribonucleoside diphosphates are added in combination.
Recall that, Nirenberg and Matthei said
that, the probability of the insertion of a specific ribonucleotide is proportional to the availability of that molecule, relative other available ribonucleotides.
If they knew the relative proportion of each type of
ribonucleoside diphosphate, they could predict the frequency of any particular triplet codon occurring in the synthetic mRNA.
Example: ! Suppose A and C are added at a ratio of 1A:5C
as three ribonucleotides, will bind to it and
attract the correct charged tRNA corresponding to the triplet code. ! The triplet codon in mRNA attracts a complimentary sequence within the tRNA
Then: there is a 1/6 chance for an A and a 5/6
chance for a C ! So: !
Possible Possible Probability of occurrence of Triplets Components any triplet AAA 3A (1/6)3= 1/216=0.4%
Final% 0.4
1C:2A
(5/6)(1/6)2=5/216=2.3% AAC ACA 3x2.3=6.9
2C:1A
ACC (5/6)2(1/6)=25/216=11.6%
3C
(5/6)3=125/216=57.9%
,CAA
CAC 3x11.6=34.8
CCA CCC
57.9
100.0
! Therefore: By examining the percentages
of any given amino acid incorporated into the protein synthesized under the direction of this message, a probable base compositions for each amino acid can be proposed. Percentage of amino acids Probable base- composition in protein Assignments Lysine <1 AAA Glutamine 2 1C:2A Asparagine 2 1C:2A Threonine 12 2C:1A Histidine 14 2C:1A 1C:2A Proline 69 CCC 2C:1A Triplet Binding Assay ! Developed by Nirenberg and Philip Leder ! The technique took advantage of the observation that ribosomes, when presented with an RNA sequence as short
Repeating Copolymers
! !
Developed by Gobind Khorona
in the early 60s Long RNA molecules are synthesized consisting of short sequences repeated many times
Wobble Hypothesis ! ! !
A pattern of degeneracy at the third position
Discovered by Crick in 1966 The first two base pairings in a codon-anticodon interaction is more critical while the third is more relaxed.
