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DNA Replication Microbial Anatomy Notes

The document discusses the DNA replication process and the key enzymes involved. It describes the initiation, elongation and termination steps of replication and the roles of DNA polymerase, helicase, ligase and other enzymes in catalyzing replication.

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Umar Amber
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28 views

DNA Replication Microbial Anatomy Notes

The document discusses the DNA replication process and the key enzymes involved. It describes the initiation, elongation and termination steps of replication and the roles of DNA polymerase, helicase, ligase and other enzymes in catalyzing replication.

Uploaded by

Umar Amber
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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DNA Replication Process - Machinery and Enzymes

We know that DNA is a self-replicating structure and DNA replicates semi-conservatively.


However, DNA replication is catalyzed by a set of enzymes. Let’s learn about the DNA
replication process and the role of enzymes involved in DNA replication.

DNA Replication

In the process of DNA replication, the DNA makes multiple copies of itself. It is a biological
polymerization, which proceeds in the sequence of initiation, elongation, and termination. It is an
enzyme-catalyzed reaction. DNA Polymerase is the main enzyme in the replication process.

DNA Replication Process

DNA Replication Steps


Following are the important steps involved in DNA replication:
Initiation

DNA replication demands a high degree of accuracy because even a minute mistake would result
in mutations. Thus, replication cannot initiate randomly at any point in DNA.

For the replication to begin there is a particular region called the origin of replication. This is the
point where the replication originates. Replication begins with the spotting of this origin
followed by the unwinding of the two DNA strands.

Unzipping of DNA strands in their entire length is not feasible due to high energy input. Hence,
first, a replication fork is created catalyzed by the helicase enzyme, which unzips the DNA
strand.

Elongation

As the strands are separated, the polymerase enzymes start synthesizing the complementary
sequence in each of the strands. The parental strands will act as a template for newly
synthesizing daughter strands.

It is to be noted that elongation is unidirectional i.e. DNA is always polymerized only in the 5′ to
3′ direction. Therefore, in one strand (the template 3‘→5‘) it is continuous, hence called
continuous replication while on the other strand (the template 5‘→3‘) it is discontinuous
replication. They occur as fragments called Okazaki fragments. The enzyme called DNA ligase
joins them later.

Termination

Termination of replication occurs in different ways in different organisms. In E.coli like


organisms, chromosomes are circular. And this happens when the two replication forks between
the two terminals meet each other.
DNA Replication Fork

Role of Enzymes in DNA Replication


DNA replication is a highly enzyme-dependent process. There are many enzymes involved in
DNA replication, which includes the enzymes, DNA-dependent DNA polymerase, helicase,
ligase, etc. Among them, DNA-dependent DNA polymerase is the main enzyme.

DNA-dependent DNA polymerase


It helps in the polymerization, catalyzes and regularizes the whole process of DNA replication
with the support of other enzymes. Deoxyribonucleoside triphosphates are the substrate as well
as the energy provider for the replication process. DNA polymerase is of three types:

DNA Polymerase I

It is a DNA repair enzyme. It is involved in three activities:

 5′-3′ polymerase activity


 5′-3′ exonuclease activity
 3′-5′ exonuclease activity

DNA Polymerase II

It is responsible for primer extension and proofreading.


DNA Polymerase III

It is responsible for in vivo DNA replication.

Helicase

Helicase is the enzyme, which unzips the DNA strands by breaking the hydrogen bonds between
them. Thus, it helps in the formation of the replication fork.

Ligase

Ligase is the enzyme which joins together the Okazaki fragments of the discontinuous DNA
strands.

Primase

This enzyme helps in the synthesis of RNA primer complementary to the DNA template strand.

Endonucleases

These produce a single-stranded or a double-stranded cut in a DNA molecule.

Single-stranded Binding Proteins

It binds to single-stranded DNA and protects it from forming secondary structures.

DNA Replication Process in Prokaryotes

The DNA replication in prokaryotes takes place in the following place:

1. The two strands of DNA unwind at the origin of replication.


2. Helicase opens the DNA and replication forks are formed.
3. The DNA is coated by the single-strand binding proteins around the replication fork to
prevent rewinding of DNA.
4. Topoisomerase prevents the supercoiling of DNA.
5. RNA primers are synthesized by primase. These primers are complementary to the DNA
strand.
6. DNA polymerase III starts adding nucleotides at the end of the primers.
7. The leading and lagging strands continue to elongate.
8. The primers are removed and the gaps are filled with DNA Polymerase I and sealed by
ligase.

DNA Replication in Eukaryotes


The DNA replication in eukaryotes is similar to the DNA replication in prokaryotes. However,
the initiation process is more complex in eukaryotes than prokaryotes. In eukaryotes, there are
multiple origins of replication present. A pre-replication complex is made with other initiator
proteins. The process is entirely the same but the enzymes used are different. E.g. in eukaryotes,
the polymerization process is carried out by the enzyme Pol δ, whereas in prokaryotes it is done
by DNA Pol III.

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