DNA translation

Protein synthesis inhibition—prokaryotes Protein synthesis inhibition—eukaryotes Mistranslation on ribosomes Nonsense mutation suppression DNA translation Phenotypic suppression Membrane leakiness Nucleic acid binding/precipitation... 

For their elucidation of DNA s secondary structure and function, Watson and Crick, along with biophysicist Maurice Wilkins (1916-2004), received the 1962 Nobel Prize in Physiology of Medicine. Subsequent research soon led to an understanding of how the nucleotide sequences in DNA translate into the synthesis of proteins. The Nobel laureates are shown in Figure 13.32. 

Birney, E., Thompson, 1. D., and Gibson, T. J. (1996). PairWise and SearchWise finding the optimal alignment in a simultaneous comparison of a protein profile against all DNA translation frames. Nucleic Acids Res. 24, 2730-2739. 

Genes are DNA and carry the inheritable characteristics of an organism and these characteristics are normally expressed at the molecular level via protein synthesis Gene expression consists of two stages transcription and translation, both of which involve RNAs Sections 28 11 and 28 12 describe these RNAs and their roles m transcription and translation... 

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... 

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... 

Transcription (Section 28 11) Construction of a strand of mRNA complementary to a DNA template Transfer RNA (tRNA) (Section 28 11) A polynucleotide of n hose that is bound at one end to a unique amino acid This ammo acid is incorporated into a growing peptide chain Transition state (Section 3 1) The point of maximum energy in an elementary step of a reaction mechanism Translation (Section 28 12) The reading of mRNA by van ous tRNAs each one of which is unique for a particular ammo acid... 

The techniques described thus far cope well with samples up to 10 kDa. Molecular mass determinations on peptides can be used to identify modifications occurring after the protein has been assembled according to its DNA code (post-translation), to map a protein structure, or simply to confirm the composition of a peptide. For samples with molecular masses in excess of 10 kDa, the sensitivity of FAB is quite low, and such analyses are far from routine. Two new developments have extended the scope of mass spectrometry even further to the analysis of peptides and proteins of high mass. 

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 types of molecules synthesized by biotechnological techniques are restricted to those biomolecules whose stmctures can be encoded in the DNA of organisms capable of translating them into functional nanomaterials. Other types of molecules and nanomaterials can be synthesized by chemical synthetic approaches, such as covalent syntheses and molecular self-assembly of molecular units. 

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. 

If an antibody to the protein of interest is available, it is sometimes possible to use vector sequences, eg, the beta-galactosidase promoter sequence, to direct the transcription of the passenger DNA into messenger RNA and the translation of that mRNA into protein which can be recognized by the antibody. Although this method is somewhat less reHable than the use of nucleic acid probes, specialized vectors are available for this purpose. 

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. 

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