ZOOHCC - 501: Molecular Biology (Theory)
Unit 2: DNA Replication
Eukaryotic genomes are much more complex and larger in size than
prokaryotic genomes. The human genome has three billion base pairs per
haploid set of chromosomes, and 6 billion base pairs are replicated during
the S phase of the cell cycle. This means that there must be multiple
origins of replication on the eukaryotic chromosome in order for all the
DNA to be replicated in a timely manner; humans can have up to 100,000
origins of replication. The rate of replication is approximately 100
nucleotides per second, much slower than prokaryotic replication.
The number of DNA polymerases in eukaryotes is much more than
prokaryotes: 14 are known, of which five are known to have major roles
during replication and have been well studied. They are known as pol α,
pol β, pol γ, pol δ, and pol ε. I won’t ever ask you the names of these
polymerases – learn the names of the prokaryotic polymerases.
Eukaryotic DNA is wound around proteins known as histones to form
structures called nucleosomes. The DNA must be made accessible in order
for DNA replication to proceed. The chromatin (the complex between DNA and
proteins) may undergo some chemical modifications, so that the DNA may be
able to slide off the histones or otherwise be accessible to the enzymes
of the DNA replication machinery. Prokaryotes do not packaged their DNA by
wrapping it around histones.
Unlike prokaryotic chromosomes, eukaryotic chromosomes are linear. As
you’ve learned, the enzyme DNA pol can add nucleotides only in the 5′ to
3′ direction. In the leading strand, synthesis continues until the end of
the chromosome is reached. On the lagging strand, DNA is synthesized in
short stretches, each of which is initiated by a separate primer. When the
replication fork reaches the end of the linear chromosome, there is no
place for a primer to be made for the DNA fragment to be copied at the end
of the chromosome. These ends thus remain unpaired, and over time these
ends may get progressively shorter as cells continue to divide.
The ends of the linear chromosomes are known as telomeres, which have
repetitive sequences that do not code for a particular gene. These
telomeres protect the genes that are located on the chromosome from
getting deleted as cells continue to divide. In humans, a six base pair
sequence, TTAGGG, is repeated 100 to 1000 times. The discovery of the
enzyme telomerase (Figure 1) helped in the understanding of how chromosome
ends are maintained. The telomerase enzyme contains a catalytic part and a
built-in RNA template. It attaches to the end of the chromosome, and
complementary bases to the RNA template are added on the 3′ end of the DNA
strand. Once the 3′ end of the lagging strand template is sufficiently
elongated, DNA polymerase can add the nucleotides complementary to the
ends of the chromosomes. Thus, the ends of the chromosomes are
replicated.
