Codons 148

Analysis of tlie global statistics of protein sequences has recently allowed light to be shed on anotlier puzzle, tliat of tlie origin of extant sequences [170]. One proposition is tliat proteins evolved from random amino acid chains, which predict tliat tlieir length distribution is a combination of the exponentially distributed random variable giving tlie intervals between start and stop codons, and tlie probability tliat a given sequence can fold up to fonii a compact... 

Codon Sequence of three nucleotides that codes for a single amino acid (or a termination signal)... 

The genetic code (Table 28 3) is the message earned by mRNA It is made up of triplets of adjacent nucleotide bases called codons Because mRNA has only four dif ferent bases and 20 ammo acids must be coded for codes using either one or two nucleotides per ammo acid are inadequate If nucleotides are read m sets of three how ever the four mRNA bases generate 64 possible words more than sufficent to code for 20 ammo acids... 

In addition to codons for ammo acids there are start and stop codons Protein biosynthesis begins at a start codon and ends at a stop codon of mRNA The start codon IS the nucleotide triplet AUG which is also the codon for methionine The stop codons are UAA UAG and UGA... 

Among the 76 nucleotides of tRNA are two sets of three that are especially important The first is a group of three bases called the anticodon, which is comple mentary to the mRNA codon for the ammo acid being transferred Table 28 3 lists two mRNA codons for phenylalanine UUU and UUC (reading m the 5 3 direction) Because base pairing requires the mRNA and tRNA to be antiparallel the two anticodons are read m the 3 5 direction as AAA and AAG... 

As described in the preceding sections protein synthesis involves transcription of the DNA to rtiRNA followed by translation of the mRNA as an amino acid sequence In addition to outlining the mechanics of transcription we have described the relationship among mRNA codons tRNA anticodons and ammo acids... 

FIGURE 28 12 Translation of mRNA to an ammo acid sequence of a protein starts at an mRNA codon for methionine Nucleophilic acyl substitution transfers the N formylmethionme residue from Its tRNA to the ammo group of the next ammo acid (shown here as alanine) The process converts an ester to an amide... 

Section 28 11 Three RNAs are involved m gene expression In the transcription phase a strand of messenger RNA (mRNA) is synthesized from a DNA tern plate The four bases A G C and U taken three at a time generate 64 possible combinations called codons These 64 codons comprise the genetic code and code for the 20 ammo acids found m proteins plus start and stop signals The mRNA sequence is translated into a prescribed protein sequence at the ribosomes There small polynucleotides called... 

Section 28 12 The start codon for protein biosynthesis is AUG which is the same as the codon for methionine Thus all proteins initially have methionine as their N terminal ammo acid but lose it subsequent to their formation The reaction responsible for extending the protein chain is nucleophilic acyl substitution... 

Anticodon (Section 27 28) Sequence of three bases in a mole cule of tRNA that is complementary to the codon of mRNA for a particular ammo acid... 

As shown in Figure 45.1, the bases appear in complementary pairs, A with T and G with C in this particular example, the sequence for one strand of DNA is A-T-C-G-T- while the other strand is -T-A-G-C-A-. The sequences of the bases attached to the sugar-phosphate backbone direct the production of proteins from amino acids. Along each strand, groups of three bases, called codons, correspond to individual amino acids. For example, in Figure 45.1, the triplet CGT, acting as a codon, would correspond to the amino acid serine. One codon, TAG, indicates where synthesis should begin in the DNA strand, and other codons, such as ATT, indicate where synthesis should stop. 

When the cell requires instructions for protein production, part of the code on DNA, starting at an initiator and ending at a stop codon, is converted into a more mobile form by transferring the DNA code into a matching RNA code on a messenger ribonucleic acid (mRNA), a process known as transcription. The decoding, or translation, of mRNA then takes place by special transfer ribonucleic acids (tRNA), which recognize individual codons as amino acids. The sequence of amino acids is assembled into a protein (see Proteins section). In summary, the codes on DNA... 

The information needed to reproduce and support an animate species is given by the order in which the nitrogen bases occur along the DNA or RNA chains (-C-T-T-A-G-, for example). A sequence of three such bases (a codon) provides the fundamental unit of information. 

Peptides and proteins are formed by linking successive amino acids into chains or rings. The order (sequence) and types of amino acids (read from the sequence of codons) determine the chemical and physical properties of peptides and proteins (and enzymes). 

A potentially general method of identifying a probe is, first, to purify a protein of interest by chromatography (qv) or electrophoresis. Then a partial amino acid sequence of the protein is deterrnined chemically (see Amino acids). The amino acid sequence is used to predict likely short DNA sequences which direct the synthesis of the protein sequence. Because the genetic code uses redundant codons to direct the synthesis of some amino acids, the predicted probe is unlikely to be unique. The least redundant sequence of 25—30 nucleotides is synthesized chemically as a mixture. The mixed probe is used to screen the Hbrary and the identified clones further screened, either with another probe reverse-translated from the known amino acid sequence or by directly sequencing the clones. Whereas not all recombinant clones encode the protein of interest, reiterative screening allows identification of the correct DNA recombinant. 

Translation of the Foreign Gene. The translation of a mRNA into a protein is governed by the presence of appropriate initiation sequences that specify binding of the mRNA to the ribosome. In addition, not all the codons of the genetic code are used equally frequently by all organisms. 

Efficient translation depends on matching the preferred pattern of host codon usage in the heterologous gene. 

Pig. 3. Representation of promoter sites on the pro-enkephalin gene. The numbers represent the distance in nucleotides from the pro-enkephalin initiation codon the arrow indicates the direction of transcription. The TATA promoter box occurs immediately before the pro-enkephalin initiation site the AP-2 site, which binds immediate-early gene products, is 70 nucleotides upstream, and the CRE site, which binds a regulatory protein involved in cAMP induction of mRNA synthesis, is 107 nucleotides upstream from the initiation codon. The expanded section shows that the CRE site actually consists of two elements, ENKCRE-1 and ENKCRE-2, which separately confer cAMP sensitivity to pro-enkephalin mRNA synthesis. 

Cellular protein biosynthesis involves the following steps. One strand of double-stranded DNA serves as a template strand for the synthesis of a complementary single-stranded messenger ribonucleic acid (mRNA) in a process called transcription. This mRNA in turn serves as a template to direct the synthesis of the protein in a process called translation. The codons of the mRNA are read sequentially by transfer RNA (tRNA) molecules, which bind specifically to the mRNA via triplets of nucleotides that are complementary to the particular codon, called an anticodon. Protein synthesis occurs on a ribosome, a complex consisting of more than 50 different proteins and several stmctural RNA molecules, which moves along the mRNA and mediates the binding of the tRNA molecules and the formation of the nascent peptide chain. The tRNA molecule carries an activated form of the specific amino acid to the ribosome where it is added to the end of the growing peptide chain. There is at least one tRNA for each amino acid. 

It has been known for some time that tetracyclines are accumulated by bacteria and prevent bacterial protein synthesis (Fig. 4). Furthermore, inhibition of protein synthesis is responsible for the bacteriostatic effect (85). Inhibition of protein synthesis results primarily from dismption of codon-anticodon interaction between tRNA and mRNA so that binding of aminoacyl-tRNA to the ribosomal acceptor (A) site is prevented (85). The precise mechanism is not understood. However, inhibition is likely to result from interaction of the tetracyclines with the 30S ribosomal subunit because these antibiotics are known to bind strongly to a single site on the 30S subunit (85). 

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