The case of the happy tree

Compound found in this

plant- cmaptothecin- has
anti cancer properties
Shown to inhibit one of
the crucial enzymes in
the process of DNA
replication- the
topoisomerase

9.1 Genetic Information Must Be

Accurately Copied Every Time a Cell
Divides
Replication has to be extremely accurate:
1 error/million bp leads to 6400 mistakes every time a cell
divides, which would be catastrophic.

Replication also takes place at high speed:

E. coli replicates its DNA at a rate of 1000 nucleotides/second.

9.2 All DNA Replication Takes Place in a

Semiconservative Manner
Proposed DNA Replication Models:
Conservative replication model
Dispersive replication model
Semiconservative replication

9.2 All DNA Replication Takes Place in a

Semiconservative Manner
Meselson and Stahls Experiment:
Two isotopes of nitrogen:
14N common form; 15N rare heavy form
E. coli were grown in a 15N media first, then transferred to 14N media
Cultured E. coli were subjected to equilibrium density gradient
centrifugation

9.2 All DNA Replication Takes Place in a

Semiconservative Manner
Modes of Replication
Replicons: Units of replication.
Replication origin

Theta replication: circular DNA, E.coli; single origin of replication

forming a replication fork, and it is usually a bidirectional
replication.

DNA Replication

Modes of Replication
In eukaryotes

12.2 All DNA Replication Takes Place in a

Semiconservative Manner
Requirements of replication
A template strand
Raw materials (substrates)
Enzymes and other proteins

12.2 All DNA Replication Takes Place in a

Semiconservative Manner
Direction of Replication
DNA polymerase add nucleotides only to the 3 end of growing
strand
The replication can only go from 5 3

9.2 All DNA Replication Takes Place in a

Semiconservative Manner
- Direction of Replication

Leading strand: undergoes continuous replication.

Lagging strand: undergoes discontinuous
replication.
Okazaki fragment: the discontinuously synthesized short DNA
fragments forming the lagging strand.

9.3 Bacterial Replication Requires a Large Number of

Enzymes and Proteins
Bacterial DNA Replication
Initiation: 245 bp in the oriC. (single
origin replicon); an initiation protein.
Unwinding

Initiator protein
DNA helicase
Single-strandbinding proteins (SSBs)
DNA gyrase (topoisomerase)

9.3 Bacterial Replication Requires a Large

Number of Enzymes and Proteins
Bacterial DNA Replication
Primers: an existing group of RNA nucleotides with a 3-OH group to which a
new nucleotide can be added. It is usually 10 ~ 12 nucleotides long.
Primase: RNA polymerase

9.3 Bacterial Replication Requires a Large

Number of Enzymes and Proteins
Bacterial DNA Replication
Elongation: Carried out by DNA
polymerase III
Removing RNA primer: DNA
polymerase I
Connecting nicks after RNA
primers are removed: DNA
ligase
Termination: when replication
fork meets or by termination
protein (Tus)

9.3 Bacterial Replication Requires a Large

Number of Enzymes and Proteins
The fidelity of DNA Replication
Proofreading: DNA polymerase I: 3 5 exonuclease activity
removes the incorrectly paired nucleotide.
Mismatch repair: correct errors after replication is complete.

9.4 Eukaryotic DNA Replication Is Similar to

Bacterial Replication but Differs in Several
Aspects
Eukaryotic DNA Replication
Eukaryotic DNA polymerase:

9.4 Eukaryotic DNA Replication Is Similar to

Bacterial Replication but Differs in Several
Aspects
Eukaryotic DNA Replication
Origin licensing (licensing factor)
The location of DNA replication within the nucleus: DNA
polymerase is fixed in location and template RNA is threaded
through it.
Replication at the ends of chromosomes:
Telomeres and telomerase
Fig 9.15