食品生物化学chapter 28-DNA replication.pptVIP

Eukaryotic Chromosomes Have Multiple replication Origins Eukaryotic chromosomes are often very long and have numerous replication origins scattered along each chromosome. Replication is bi-directional, as in bacteria. A pair of replication forks starts at each origin of replication and the two forks then move in opposite directions There are estimated to be between 10,000 and 100,000 replication origins in a dividing human somatic cell. Synthesis of Eukaryotic DNA The synthesis of DNA in eukaryotes is less well investigated than in bacteria. But synthesis of eukaryotic DNA resembles that in bacteria in most general respects: a) it is semi-conservative and bidirectional b) semi-discontinuous, one strand is made in short fragments c) RNA primers are needed to start new strands. In animal cells, two DNA polymerases (? and δ-Delta ) are involved in chromosome replication. DNA polymerase ? is responsible for initiation of new strands. DNA polymerase δ is responsible for elongation of new strands. Eukaryotic DNA Replication A) The process begins with the binding of a primase which produces an RNA primer. Subsequently, DNA polymerase ? binds and initiates a short segment of DNA called initiator DNA (iDNA, only 3~4 base long). B) Replication factor C associates with the iDNA and is C) helpful in positioning DNA polymerase δ. D) DNA polymerase δ elongates the new DNA strand. Linking of the Okazaki fragments differs significantly between animal and bacterial cells. In animals, there is no equivalent [?kw?v(?)l(?)nt] of the dual function DNA polymerase I of bacteria. The RNA primers are removed by an exonuclease (MF1) and the gaps are filled by the DNA polymerase δ that is working on the lagging strand. As in bacteria, the nicks are sealed by DNA ligase. The 5’-End is Potentially Lost in Replication of Linear DNA When an RNA primer is removed after initiating a strand of linear DNA the gap cannot be filled by DNA as there is no strand for DNA polymerase to elongate. Thus,