Microbiology

During the 1950s and 1960s it became apparent that DNA is essential in the synthesis of proteins. Proteins are used as structural materials in the cells and function as enzymes. In addition, many specialized proteins function in cellular activities. For example, in bacteria, flagella and pili are composed of protein.

The genetic code. The key element of a protein molecule is how the amino acids are linked. The sequences of amino acids, determined by genetic codes in DNA, distinguish one protein from another. The genetic code consists of the sequence of nitrogenous bases in the DNA. How the nitrogenous base code is translated to an amino acid sequence in a protein is the basis for protein synthesis.

In order for protein synthesis to occur, several essential materials must be present. One is a supply of the 20 amino acids which make up most proteins. Another essential element is a series of enzymes that will function in the process. DNA and another form of nucleic acid called ribonucleic acid (RNA) are also essential. RNA carries instructions from the nuclear DNA into the cytoplasm, where protein is synthesized. RNA is similar to DNA, with three exceptions. First, the carbohydrate in RNA is ribose rather than deoxyribose. Second, RNA nucleotides contain the pyrimidine uracil rather than thymine. And third, RNA is usually single-stranded.

Types of RNA. In the synthesis of protein, three types of RNA are required. The first is called ribosomal RNA (rRNA) and is used to manufacture ribosomes. Ribosomes are ultramicroscopic particles of rRNA and protein where amino acids are linked to one another during the synthesis of proteins. Ribosomes may exist along the membranes of the endoplasmic reticulum in eukaryotic cells or free in the cytoplasm of prokaryotic cells.

A second important type of RNA is transfer RNA (tRNA), which is used to carry amino acids to the ribosomes for protein synthesis. Molecules of tRNA exist free in the cytoplasm of cells. When protein synthesis is taking place, enzymes link tRNA to amino acids in a highly specific manner.

The third form of RNA is messenger RNA (mRNA), which receives the genetic code from DNA and carries it into the cytoplasm where protein synthesis takes place. In this way, a genetic code in the DNA can be used to synthesize a protein at a distant location at the ribosome. The synthesis of mRNA, tRNA, and rRNA is accomplished by an enzyme called RNA polymerase.

Transcription. Transcription is one of the first processes in the overall process of protein synthesis. In transcription, a strand of mRNA is synthesized using the genetic code of DNA. RNA polymerase binds to an area of a DNA molecule in the double helix (the other strand remains unused). The enzyme moves along the DNA strand and selects complementary bases from available nucleotides and positions them in an mRNA molecule according to the principle of complementary base pairing (Figure 1 ). The chain of mRNA lengthens until a stop code is received. 

Figure 1

The synthesis of mRNA using a strand of DNA as a template.

The nucleotides of the DNA strands are read in groups of three. Each triplet is called acodon. Thus, a codon may be CGA, or TTA, or GCT, or any other combination of the four bases, depending on their sequence in the DNA strand. The mRNA molecule consists of a series of codons received from the genetic message in the DNA.

Once the stop codon has been reached, the mRNA molecule leaves the DNA molecule, and the DNA molecule rewinds to form a double helix. Meanwhile, the mRNA molecule proceeds thorough the cellular cytoplasm toward the ribosomes.

Translation. Translation is the process in which the genetic code will be “translated” to an amino acid sequence in a protein. The process begins with the arrival of the mRNA molecule at the ribosomes. While mRNA was being synthesized, tRNA molecules were uniting with their specific amino acids according to the activity of specific enzymes. The tRNA molecules then began transporting their amino acids to the ribosomes to meet the mRNA molecule.

After it arrives at the ribosomes, the mRNA molecule exposes its bases in sets of three, the codons. Each codon has a complementary codon called an anticodon on a tRNA molecule. When the codon of the mRNA molecule complements the anticodon on a tRNA molecule, the latter places the particular amino acid in that position. Then the next codon of the mRNA is exposed, and the complementary anticodon of a tRNA molecule matches with it. The amino acid carried by the second tRNA molecule is thus positioned next to the first amino acid, and the two are linked. At this point, the tRNA molecules release their amino acids and return to the cytoplasm to link up with new molecules of amino acid.

The ribosome then moves farther down the mRNA molecule and exposes another codon which attracts another tRNA molecule with its anticodon. Another amino acid is brought into position. In this way, amino acids continue to be added to the growing chain until the ribosome has moved down to the end of the mRNA molecule. The sequence of codons on the mRNA molecule thus determines the sequence of amino acids in the protein being constructed (Figure 2 ). 

Figure 2

Steps in the synthesis of protein beginning with the genetic code in DNA and ending with the finished polypeptide chain.

Once the protein has been completely synthesized, it is removed from the ribosome for further processing. For example, the protein may be stored in the Golgi body of a eukaryotic cell before release, or a bacterium may release it as a toxin. The mRNA molecule is broken up and the nucleotides are returned to the nucleus. The tRNA molecules return to the cytoplasm to unite with fresh molecules of amino acids, and the ribosome awaits the arrival of a new mRNA molecule.

Gene control. The process of protein synthesis does not occur constantly in the cell, but rather at intervals followed by periods of genetic “silence.” Thus, the process of gene expression is regulated and controlled by the cell.

The control of gene expression can occur at several levels in the cell. For example, genes rarely operate during mitosis. Other levels of gene control can occur at transcription, when certain segments of DNA increase and accelerate the activity of nearby genes. After transcription has taken place, the mRNA molecule can be altered to regulate gene activity. For example, it has been found that eukaryotic mRNA contains many useless bits of RNA that are removed in the production of the final mRNA molecule. These useless bits of nucleic acid are called introns. The remaining pieces of mRNA, called exons, are then spliced to form the final mRNA molecule. Bacterial mRNA lacks introns.

The concept of gene control has been researched thoroughly in bacteria. In these microorganisms, genes have been identified as structural genes, regulator genes, and control regions. The three units form a functional unit called the operon.

The operon has been examined in close detail in certain bacteria. It has been found that certain carbohydrates can induce the presence of the enzymes needed to digest those carbohydrates. For example, when lactose is present, bacteria synthesize the enzymes needed to break it down. Lactose acts as an inducer molecule in the following way: In the absence of lactose, a regulator gene produces a repressor protein, which binds to a control region called the operator site. This binding prevents the structural genes from encoding the enzyme for lactose digestion. When lactose is present, however, it binds to the repressor protein and thereby removes the repressor at the operator site. With the operator site free, the structural genes are released to produce their lactose-digesting enzyme.