Molecular Genetics

DNA replication-3 and transcription

Lecture 4 (08-08-2014)

DNA Replication
The prokaryotic replication bubble.
In E. coli and other bacteria, the DNA double helix separates at the origin of replication (yellow
star), creating a replication bubble. Synthesis of the new strands proceeds in both directions
away from the origin of replication, forming a replication bubble, and the place where the two
strands separate is called the replication fork. As replication proceeds, the replication bubble gets
larger, so that eventually the replication forks meet, and the two new chromosomes separate.
Principles of Biology
It takes E. coli about 40 minutes to
duplicate its genome of 4.6 10
6

nucleotide pairs.
DNA Replication
Initiation of Replication.
During the initiation of DNA replication, DNA helicase unwinds the double helix and continues
to break the hydrogen bonds that hold the two parental strands together. Single-strand DNA
binding proteins (SSBs) bind to the single-stranded DNA to prevent spontaneous hydrogen
bonding of the single strands to each other. Topoisomerase stabilizes the region directly
ahead of the replication fork by breaking the strands, turning them, and rejoining them to
relieve the structural strain created by the unwinding of the double helix..
Principles of Biology
Figure 5-27 Molecular Biology of the Cell ( Garland Science 2008)
DNA Replication
Initiation of Replication in E coli.
Figure 5-28 Molecular Biology of the Cell ( Garland Science 2008)
Methylation of the E. coli replication origin creates a refractory period for DNA initiation.
DNA methylation occurs at GATC sequences, 11 of which are found in the origin of
replication (spanning about 250 nucleotide pairs). About 10 minutes after replication the
hemimethylated origins become fully methylated by a DNA methylase enzyme.
DNA Replication
DNA Replication is tightly controlled
The only point at which E. coli can control DNA replication is initiation
DNA Replication
Termination of replication in prokaryotes
Replication termini in E. Coli are located in a region between two set of ter sites (23 bp long).
Each set of ter sites is direction specific, one set allows replication fork in but the other set
cant let it pass. Thus replication fork is trapped at termination site.
Not applicable in eukarayotes as there are multiple replication bubbles.
Figure 13.30 Gene XI (JBL 2014))
ter sequences
DNA Replication
Eukaryotic DNA Replication
Because of the linear nature of eukaryotic DNA,
DNA polymerase III cannot add the final
sequence of DNA to the 5' end of the lagging
daughter strand.
In most eukaryotes, chromosomes are linear,
so there can be multiple replication bubbles at
any given point in the replication process.
To prevent the loss of genetic information,
eukaryotic DNA has special end sequences
called telomeres, which do not code for
proteins.
Telomerase is an enzyme that contains an RNA
template that it uses to lengthen the telomere of
the template strand. Lengthening the template
strand allows the functional part of the daughter
strand to be completed.
Principles of Biology
Figure 5-40 Molecular Biology of the Cell ( Garland Science 2008)
DNA Replication
Replication of telomeres in eukaryotes.
Principles of Biology
DNA Replication
Figure 8 Replication of telomeres in eukaryotes.
The enzyme telomerase extends the end of the parental strand using its own RNA template. A
new RNA primer complementary to this extension allows the daughter strand to be completed.
Principles of Biology
Telomeres in Aging and Cancer
http://learn.genetics.utah.edu/content/begin/traits/telomeres/
Telomeres in Aging and Cancer
Also checkout interesting readings:
http://www.popsci.com/science/article/2011-07/man-who-would-stop-time
http://www.scientificamerican.com/article.cfm?id=telomeres-telomerase-and

Watch Flash animation at
http://learn.genetics.utah.edu/content/begin/traits/telomeres/