3-DNA的复制分子生物学幻灯片.pptVIP

3. 真核细胞DNA的复制调控 （1）细胞生活周期水平调控 DNA复制只发生在S期 （2）染色体水平调控 不同部位的复制子按一定的时间顺序在S期起始复制 （3）复制子水平调控 * It remained conceivable that DNA replication might follow a dispersive model, so that both strands of DNA in the chromosomes of cells after one generation on 14N might be intermediate in density. Meselson and Stahl eliminated this possibility by the following experiment: DNA isolated from 15N-labeled cells kept one generation on 14N was heated at 100°C so that the DNA duplexes were denatured into their component single strands. When this heat-denatured DNA was analyzed by CsCl density gradient ultracentrifugation, two distinct bands were observed, showing that one strand was 15N-labeled and the other strand was 14N-labeled. A dispersive mode of replication would have yielded a single band of DNA of intermediate density. An interesting feature of density gradient ultracentrifugation is that the width of the band occupied by a macromolecular species in the gradient is inversely proportional to the square root of the molecular weight of the macromolecule. The band widths of the two bands seen with heat-denatured DNA both correspond to molecular masses one-half that of native DNA. This experiment established that DNA replication proceeded by a semiconservative mechanism and also verified that DNA was indeed composed of two polynucleotide strands of equal size. * * Topoisomerases can both catenate and decatenate circular DNA molecules. Circular DNA molecules are said to be catenated if they are linked together lib two rings of a chain (Figure 6-23a). Of these two activities, the ability of topoisomerases to decatenate DNA is of clear bim logical importance. As we will see in Chapter 8, catenated DNA molecules are commonly produced as a round of DNA replication is 6nished (see Figure 8-33). Topoisomerases play the essential role of unlinking these DNA molecules to allow them to separate into the two daughter cells for cell division. Decatenation of two covalently closed circu