[Q2(A) Discuss in brief the process of DNA replication in eukaryotes along with structure and function of

eukaryotic DNA polymerases and also trace out how prokaryotic DNA replication different from
eukaryotic replication.]

 DNA replication in eukaryotes

DNA replication in eukaryotes is the process by which the DNA is synthesized or
doubled in the S phase of cell cycle. Its mechanism is as follows
 DNA replicating tools
in eukaryotic cells following is the tool kit required for DNA replication,
 ORC: that is origin Recognizing proteins
 top isomerases I/II
 DNA helicase
 replication factor A that is RPA or SSB
 RFC replication factor C
 polymerase Delta and epsilon
 Polymerase Alpha
 proliferating cell nuclear antigen [PCNA]
 Primase
 DNA ligases
 FEN-1 for filling of gaps
 T antigen
 Mechanism
 first of all there will be formation of replication fork at the origin site of
DNA after ORC proteins locate the origin of replication.
 replication Fork will form and progress when DNA will unwind with the
help of proteins.
 [Karp,2011]
 the topisomerases stress down the supercoiling present in the parental DNA
complex. Then T antigen Using its DNA binding domain Produce a multi
subunit complex with the help of ATP utilization and local unwinding
occurs.
 this unwinding is further supported by RFA or SSB proteins which will
maintain DNA in this unwinding state.
 like prokaryotes, replication or synthesis of DNA occurs in different strands
that is leading strand and lagging strand. Polymerase epsilon Replicate the
leading strand in continuous fashion After primase add the primers with the
assistance of polymerase Alpha. Polymers Alpha is thought to have role with
primase.
 polymerase Delta will synthesize DNA in the form of legging strand. The
PCNA will act as sliding clamp for both polymerases allowing to move
progressively on template.
 the PCNA is loaded by proteins RFC called PCNA loaders. in this way
RNA-DNA primer is formed. polymers Alpha is substituted by polymerase
Delta which will synthesize okazaki fragments.
 the primer is displaced from okazaki fragment by FeN1 endonucleases and is
sealed with other fragments by DNA ligase. In this way replication is
completed.

[slideshare.net]
  structure and function of eukaryotic DNA polymerase
following are 5 different DNA polymerases found in eukaryotes,
 DNA polymerase Alpha
it is the largest polymerase having iron-sulphur clusters with high molecular
weight. It is also known as cytoplasm polymerase because it is also present in the
cytoplasm. It has primase catalytic activity in DNA replication.
 DNA polymerase beta
it is only present in vertebrates. A rather small size polymerase. It consists of 2
domains each having different enzymatic activity. Its function is reported in DNA
repairing system.
 DNA polymerase Gemma
this is the mitochondrial polymerase causing mitochondrial DNA replication. It is a
heterotrimeric unit consisting of 15 turns 52 beta strands and 39 Alpha helices
having catalytic site.
 DNA polymerase Delta
it is composed of 4 subunits that is POLD1, POLD 2, POLD 3, POLD 4. It is
present in mammalian cells. Its function is reported in Replication of both leading
and lagging strand. It shows its activity in the presence of proliferating cell nuclear
antigen PCNA.
 DNA polymerase epsilon
it consists of 4 subunits POLE, POLE2, POLE3 and POLE 4. POLE1 is the main
catalytic site. It is recently reported to have Major role in replication of leading
strand in continuous fashion. It also functions in DNA repair system.
  Difference between Prokaryotic and eukaryotic replication
Prokaryotic replication Eukaryotic replication
1. prokaryotic cells exhibit only 1. eukaryotic cells exhibit more
one point of origin usually origin points in their DNA due to
because their DNA is rather larger DNA size.
small that is 25 times less than 2. eukaryotic replication occurs in
that of eukaryotic DNA. nucleus of the cell.
2. as prokaryotes don't have any 3. eukaryotes possess 4 or more
nucleus their replication occurs polymerases that is polymerase
in cytoplasm. Alpha, polymerase beta,
3. prokaryotic cells have less polymerase gamma polymerase
number of polymerase than Delta, polymerase epsilon At
eukaryotes. cetera.
4. their replication rate is much 4. the replication rate of eukaryotes
faster. is very slow as compared to
 they have larger size okazaki prokaryotes due to larger
fragments usually ranging from genome.
1000 to 2000 long nucleotides. 5. they have smaller okazaki
fragments ranging from 150 to
200 nucleotides.

References
 Varma and Agarwal, cell biology, genetics, molecular biology, S Chand
publishers, 2005
 Gerald Karp, cell and molecular biology, 6th edition 2011
 Snusted and Simmons, principle of genetics,6th edition, 2012
 BENJAMIN A. PIERCE, Genetics a conceptual approach
 https://en.wikipedia.org/wiki/Eukaryotic_DNA_replication
[Q2(C) Describe the different kinds of gene mutations and justify your explanation with the
reference of induced mutations by mutagens.]

 Gene mutations
Gene mutations are defined as changes in genes or DNA sequences of an
Organism. These changes can be both qualitative and quantitative. Changes in
DNA sequence means changes in Messenger RNA sequence and corresponding
proteins. Mutations can be spontaneous as in most cases and can also be induced
by artificial means.
the gene mutation can be categorised into 2 general categories
1. point mutation
2. Multiple or gross mutations
 point mutations
whenever changes occur at a small point that is single nucleotide or nucleotide pair
is termed as point mutations. following are some types of point mutation
 deletion mutations
when some part of DNA is lost for example in a triplet codon is called as deletion
mutation.
 insertion mutation
In this type of mutation extra nucleotide got inserted in the already present DNA
sequence.
 frameshift mutation
the incorrect reading of DNA sequence which occurs after some part is lost that is
deleted or some part is inserted is called frameshift mutation. These incorrect
readings cause in the incorrect Messenger RNA sequence and inactive or defective
protein.
 base substitutions in this type of gene mutation one of the nucleotide is
substituted by another nucleotide in a triplet is called base substitution. Base
substitution Usually occurs in a triplet codon Causing a single nucleotide
substitution which in result causes change of only one amino acid in a
resulting protein which altogether changes the function of protein sometimes
and are of great genetic importance. For example in sickle cell anaemia.
  transition
In this type of substitution A purine is substituted by another purine in a triplet
that is adenine by Guanine or a pyrimidine is substituted by another pyrimidine
Is called transition Substitution.
 transversion
In this transversion substitution a purine is substituted by a pyrimidine And vice
versa. for example, adenine by thymine it is called as transverse substitution.

[Lardbucket.org]

 multiple mutations
when mutations occur in more than one nucleotide pairs or in an entire gene is
called as multiple mutations or gross mutations. Multiple mutations are actually
rearrangement of genes. There are following types,
translocation
When gene is shifted to another non homologous chromosome it is called
translocation.
inversion
when gene roam about in the same chromosome it is called As inversion mutation.

 Induced mutations
the mutations which are induced artificially through different agents is called as
induced mutations. these agents are known as mutagens. Mutagens can be
chemicals, radiations at cetera. We'll see how they play role in gene mutations.

 Induced mutations through chemicals

 nitrous acid
nitrous acid causes the deamination of the DNA base causing point transitional
mutations. In deamination the amino group that is NH2 is substituted by hydroxyl
group that is OH by nitrous acid. due to this normal adenine is converted to
hypoxanthine.

[Genes.org]
  hydroxylamine
amino group of cytosine base undergo hydroxylation And changes cytosine to
hydroxylcytosine causing point mutations at GCAT.
 Hydrazine
hydrazine destroys the rings of pyrimidine by converting them to Pyrazolone.
 Base analogous
certain chemicals have chemical similarity with the nitrogenous bases. So during
replication Of DNA instead of actual nitrogenous bases these base analogous got
picked up and joint in DNA causing mutation. For example 5 bromouracil is
similar to thymine and substituted in place of thymine.
 Radiations
radiations are of 2 types
ionizing radiation Which causes the ionization of Atom in DNA molecule such as
X Rays gamma Rays beta Rays electrons at cetera.
Non-ionising radiations causes mutations by the absorbance of energy at high
wavelength and resulting changes in that high energy parts and mutations. Such as
ultraviolet Rays the most effective wavelength is 2600 A* best absorbed by DNA.
For example, xeroderma pigmentosum.

References
 Varma and Agarwal, cell biology, genetics, molecular biology, S Chand
publishers, 2005
 Gerald Karp, cell and molecular biology, 6th edition 2011
 Snusted and Simmons, principle of genetics,6th edition, 2012
 BENJAMIN A. PIERCE, Genetics a conceptual approach
 https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/induced-
mutation