1/12 Prof. D. Figurski Microbiology Transcriber: Ogan Gurel Wednesday, 9 March 1988 10:00 AM POINT MUTATION The minimum genetic change INTRODUCTION Yesterday's lecture introduced the concept that bacteria can undergo stable hereditary genetic variation and that this variation enhances the ability of bacterial populations to acquire new characteristics. This genetic variation occurs within a single cell through stable changes in its DNA that can be passed on to all the progeny (Fig. 1)--these changes are referred to as mutations. A | mm stamue Ae LON HERITABLE Ae Ae FIGURE 1 Baceril genetics depends onthe analy of ‘able and erable sons. Mutations are characterized by the fact that they ar 1. spontaneous, 2. random and 3. occur at a relatively low frequency. In analyzing a particular gene one can expect to find a mutation in such a gene in the range of 10-6 to 10-8 cells. However, bacteria grow rapidly with a very short generation time so that this low frequency of mutation becomes quite significant in absolute time. As mentioned yesterday, there are essentially two ways in which these changes can take place. These are via: 1. point mutation, and 2. gross DNA rearrangements. Today's lecture is concerned with how point mutations result in genetic variation.II. 2/12 A HYPOTHETICAL OPERON Point mutations are essentially single nucleotide changes in DNA. Such mutations constitute the minimum DNA change that may affect the function of a gene. A hypothetical operon (Fig. 2) serves as a good model for studying the different ways in which a gene's function may be altered. oat ee = (0 Opera A B FIGURE 2: Hypotedeal peo There are six important components to this operon First, there is a promoter (P) to which the RNA polymerase binds and initiates transcription. Close to or overlapping the promoter is an operator (0) which provides for regulation of the operon through the binding of regulatory proteins. Third, the ribosome binding site (RB) controls the translational efficiency of the mRNA In addition, the operon encodes two structural genes (A and B); the translation of each gene begins and ends at its respective start (ATG) and stop codons (TAG) The promoter, operator, and ribosome binding sites can be described as signals that control the expression of the operon. The individual genes along with their respective start and stop codons comprise the structural genes of the operon. Thus, there are two general ways in which a point mutation can affect the functioning of an operon These are: 1. through mutation in a signal for expression 2. through mutation in a structural gen3/12 III. MUTATIONS IN EXPRESSION SIGNALS Iv. A. Mutations in the promoter. Such alterations can affect the interaction of RNA polymerase with the DNA. These mutations can either block or reduce the affinity of RNA polymerase binding resulting either in the elimination or reduction of transcription, respectively. Also, changes that enhance the binding of RNA polymerase may increase the transcriptional efficiency. Mutations in the operator. The operator is generally a target for a regulatory protein--in some cases this may be a repressor. A point mutation that prevents repressor-operator binding would cause constitutive expression of the gene while an alteration that enhances binding would make it difficult to turn on the operon. Mutations in the ribosome binding site. Some mutations in RB would decrease the efficiency of ribosome binding and thus reduce the efficiency of translation; the end result would be less protein. Likewise, an RB change that increases ribosome binding would produce more protein. MUTATIONS IN STRUCTURAL GENES Structural genes represent the information that is to be translated into protein. There are three types of mutations in structural genes: missense mutations, nonsense mutations, and frameshift mutations. A. issense mutations Figure 3 depicts a missense mutation in which a T is converted to a C in the DNA. The corresponding codon in the mRNA is accordingly changed so that the resulting protein product contains a proline instead of a leucine at one position in the sequence.