Benjamin A.

Pierce

•GENETICS ESSENTIALS
•Concepts and Connections
• SECOND EDITION

CHAPTER 7
DNA Replication and Recombination

© 2013 W. H. Freeman and Company

 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
7.1 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.
 7.2 ALL DNA REPLICATION TAKES PLACE IN A
SEMICONSERVATIVE MANNER (MÔ HÌNH BÁN BẢO THỦ)

PROPOSED DNA REPLICATION MODELS:

 Meselson and Stahl’s 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
 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
 Requirements of replication

• A template strand
• Raw materials
(substrates)
• Enzymes and
other proteins
 Direction of Replication

•DNA polymerase add nucleotides only to the 3 end of growing strand

•The replication can only go from 5  3
 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.
7.3 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-strand–binding
proteins (SSBs)
• DNA gyrase
(topoisomerase)
• 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
 • 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.
 7.4 Eukaryotic DNA Replication Is Similar to
Bacterial Replication but Differs in Several Aspects

• Eukaryotic DNA Replication: Eukaryotic DNA polymerase:

 7.4 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
Figure 7.15
The enzyme
telomerase is
responsible for the
replication of
chromosome ends.
Figure 7.15
The enzyme
telomerase is
responsible for the
replication of
chromosome ends.
Figure 7.15
The enzyme
telomerase is
responsible for
the replication of
chromosome
ends.