DNA Transcription in Bacteria

As with the establishment of the genetic code and information on the molecular mechanism of DNA replication (Chap. 16), the present detailed knowledge of the mechanism of DNA transcription to produce RNA rests largely upon studies with bacteria, particularly E. coli. It is convenient to treat transcription in bacteria first. 

Most of the DNA sequences which are transcribed give rise to mRNA, which is subsequently translated into protein. However, the most abundant species of RNA are ribosomal RNA (rRNA) and transfer RNA (tRNA), which do not code for protein but function in the process of translation. They are formed by a high level of transcription of a relatively small number of genes (called rRNA and tRNA genes). In bacteria, transcription of all genes is brought about by the enzyme RNA polymerase. 

Question What is the nature of the chemical reaction catalyzed by RNA polymerase The overall reaction is 

it uses the four ribonucleoside triphosphates (ATP, GTP, UTP, and CTP) to assemble an RNA chain, the sequence of which is determined by the template strand of DNA. Nucleotide addition occurs sequentially, the phosphodiester bond being formed through the same mechanism as described for DNA polymerase (see Chap. 16, Fig. 16-9). RNA chain growth is in the 5 — 3 direction. An important distinction between RNA polymerase and DNA polymerase, however, is the ability of the former to start a new chain de novo i.e., it does not have an obligatory requirement for a primer. The first nucleotide to be incorporated into the chain of RNA contains either adenine or guanine and retains its 5 triphosphate. 

Question Within a chromosome, a single DNA molecule contains a multitude of genes, and when a particular one is to be transcribed, the RNA polymerase must know where to start. How is this 

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Related topics DNA replication in bacteria (F3) Transcription in DNA replication in eukaryotes (F4) prokaryotes (G2) RNA structure (Gl) Transcription in eukaryotes an overview (G5)... 

The conversion of the information in DNA into proteins begins in the nucleus of cells with the synthesis of mRNA by transcription of DNA. In bacteria, the process begins when RNA polymerase recognizes and binds to a promoter... 

The reaction of X with S must be fast and reversible, close to if not at equilibrium with concentration of S. It can be that there is an intermediate step in which X binds to a protein kinase (a protein which phosphorylates other proteins mostly at histidine residues in bacteria) using phosphate transferred from ATP. It then gives XP which is the transcription factor, where concentration of S still decides the extent of phosphorylation. No change occurs in DNA itself. Here equilibrium is avoided as dephosphorylation involves a phosphatase, though changes must be relatively quick since, for example, cell cycling and division depend on these steps, which must be completed in minutes. We have noted that such mechanical trigger-proteins as transcription factors are usually based on a-helical backbones common to all manner of such adaptive conformational responses (Section 4.11). 

Inhibition of DNA gyrase, an enzyme required for DNA replication, transcription, repair and recombination in bacteria... 

FIGURE 27-28 Coupling of transcription and translation in bacteria. The mRNA is translated by ribosomes while it is still being transcribed from DNA by RNA polymerase. This is possible because the mRNA in bacteria does not have to be transported from a nucleus to the cytoplasm before encountering ribosomes. In this schematic diagram the ribosomes are depicted as smaller than the RNA polymerase. In reality the ribosomes (Mr 2.7 X 105) are an order of magnitude larger than the RNA polymerase (Mr 3.9 X 105). 

In bacteria, genes that encode products with interdependent functions are often clustered in an operon, a single transcriptional unit. Transcription of the genes is generally blocked by binding of a specific repressor protein at a DNA site called an operator. Dissociation of the repressor from the operator is mediated by a specific small molecule, an inducer. These principles were first elucidated in studies of the lactose (lac) operon. The Lac repressor dissociates from the lac operator when the repressor binds to its inducer, allolactose. 

A major goal in recombinant DNA technology is the production of useful foreign proteins by bacteria, yeast, or other cultured cells. Protein synthesis depends upon both transcription and translation of the cloned genes and may also involve secretion of proteins from the host cells. The first step, transcription, is controlled to a major extent by the structures of promoters and other control elements in the DNA (Chapter 28). Since eukaryotic promoters often function poorly in bacteria, it is customary to put the cloned gene under the control of a strong bacterial or viral X promoter. The latter include the X promoter PL (Fig. 28-8) and the lac (Fig. 28-2) and trp promoters of E. coli. These are all available in cloning vehicles. 

There are other types of transcriptional activators in bacteria. One is transcription factor 1 (TF1) encoded by a Bacillus subtilis phage. It is a member of the protein HU family (Chapter 27). However, unlike the nonspecific HU it binds to some sites specifically and activates transcription.143 The E. coli Ada protein is the acceptor protein in removal of methyl groups from DNA (Chapter 27). The same protein is an inducer of transcription of DNA repair enzymes in the large ada regulon. Methylation of Cys 69 of the Ada protein itself converts it into a gene activator.144... 

In bacteria transcription and translation are closely linked. Polyribosomes may assemble on single DNA strands as shown in Fig. 28-5. It has often been assumed that RNA synthesis occurs on loops of DNA that extend out into the cytosol. However, recent studies indicate that most transcription occurs in the dense nucleoid and that assembly of ribosomes takes place in the cytosol.2683 In a similar way eukaryotic transcription occurs in the nucleus and protein synthesis in the cytosol. Nevertheless, some active ribosomes are present in the nucleus.26813... 

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