11.2 – DNA Replication
Learning Objectives
- Explain the meaning of semiconservative DNA replication
- Explain why DNA replication is bidirectional and includes both a leading and lagging strand
- Explain why Okazaki fragments are formed
- Describe the process of DNA replication and the functions of the enzymes involved
- Identify the differences between DNA replication in bacteria and eukaryotes
- Explain the process of rolling circle replication
The elucidation of the structure of the double helix by James Watson and Francis Crick in 1953 provided a hint as to how DNA is copied during the process of replication. Separating the strands of the double helix would provide two templates for the synthesis of new complementary strands, but exactly how new DNA molecules were constructed was still unclear. In one model, semiconservative replication, the two strands of the double helix separate during DNA replication, and each strand serves as a template from which the new complementary strand is copied; after replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. There were two competing models also suggested: conservative and dispersive, which are shown in Figure 11.4.
Microbial Informatics and Experimentation 3 no. 1 (2013):2.
Diagram showing 3 models of DNA replication. In the conservative model the original double helix produces two double helices; one of which has two of the parent strands and one of which has two of the new strands. Another round produces 4 helices; one of which has two of the parent strands and three of which have all new strands. In semiconservative replication the first round leads to two double helices each with one old strand and one new strand. The next round leads to four double helices; two of these have an old and a new strand and two have all new strands. In dispersive replication each new round of replication results in strands with random bits from the parent strand and random bits of new strands.
Matthew Meselson (1930–) and Franklin Stahl (1929–) devised an experiment in 1958 to test which of these models correctly represents DNA replication (Figure 11.5). They grew E. coli for several generations in a medium containing a “heavy” isotope of nitrogen (15N) that was incorporated into nitrogenous bases and, eventually, into the DNA. This labeled the parental DNA. The E. coli culture was then shifted into a medium containing 14N and allowed to grow for one generation. The cells were harvested and the DNA was isolated. The DNA was separated by ultracentrifugation, during which the DNA formed bands according to its density. DNA grown in 15N would be expected to form a band at a higher density position than that grown in 14N. Meselson and Stahl noted that after one generation of growth in 14N, the single band observed was intermediate in position in between DNA of cells grown exclusively in 15N or 14N. This suggested either a semiconservative or dispersive mode of replication. Some