REPLIKASI DNA &

MUTASI GEN
BIOTEKNOLOGI FARMASI ISTN - 2017
REPLIKASI DNA
Replikasi DNA
Replikasi merupakan proses pelipatgandaan DNA
agar semua sel turunan memiliki informasi
genetik yang sama.

Ada 3 model replikasi DNA:

a) Konservatif
b) Semi konservatif
c) Dispersif
 a) Conservative Replication
Pada replikasi konservatif, kedua
molekul rantai DNA berperan sebagai
cetakan (template) untuk molekul
DNA yang baru
Molekul DNA induk (original DNA
molecule) is fully conserved during
replication

50% molekul DNA baru, dan 50%

molekul DNA induk

75% molekul DNA baru, dan 25%

molekul DNA induk
b) Dispersive Replication
Pada replikasi dispersif, kedua rantai nukleotida terpotong-potong
menjadi fragmen-fragmen yang berperan sebagai cetakan
(templates) untuk sintesis fragmen DNA baru
Potongan fragmen-fragmen tersebut akan dirakit menjadi dua
molekul DNA baru.
Pada model ini, tiap molekul DNA baru yang dihasilkan berselang-
seling dengan rantai induk.
Molekul DNA induk tidak terkonservasi.

 Dispersive replication
would always produce
hybrid molecules,
containing some original
and some new DNA, but
the proportion of new
DNA within the
molecules would
increase with each
replication event.
c) Semi-Conservtive Replication

Tiap rantai DNA berperan sebagai

template untuk sintesis rantai
DNA yang baru.

Spesifitas dari pasangan basa (A-T

; G-C) juga menegaskan bahwa
model ini akan menghasilkan
rantai DNA baru yang identik
dengan induk nya (DNA asal)
 Komponen komponen Replikasi DNA
Mulai pada origin of replication (ori)
Primase (RNA polimerase)
Helikase (pembuka untai)
DNA polimerase
Protein pengikat DNA untai tunggal (ssbp)
 Enzim Helicase adalah enzim yang berfungsi mengurai pilinan heliks
dengan memotong ikatan hydrogen antar basa untai ganda DNA
sehingga terpisah menjadi 2 untai tunggal DNA.

Single Strand Binding Protein (SSBP) adalah protein yang berfungsi

melindungi untai tunggal DNA agar tidak bergabung kembali setelah
dipisahkan oleh helicase (menstabilkan untai tunggal DNA).

DNA Primase adalah enzim untuk sintesis RNA primer dalam mengawali
pembentukan DNA baru pada leading strand atau DNA fragmen Okazaki
pada lagging strand oleh DNA Polimerase.
 DNA Polimerase adalah enzim yang memanjangkan rantai DNA baru
dengan cara membentuk ikatan fosfodiester yang merangkaikan C 5 dari
suatu nukleotida ke C 3 nukleotida yang lain. Karena fungsinya
memanjangkan dalam sintesis untai DNA, maka enzim ini memerlukan
RNA primer sebagai awalan.

Ligase adalah enzim yang berfungsi menyambungkan fragmen-

fragmen
DNA menjadi rantai yang lebih panjang.
 REPLIKASI DNA

= DUPLIKASI DNA

Jalannya replikasi DNA dari 5 ke 3 (5:3).

 Fragmen Okazaki adalah DNA pendek yang terbentuk di lagging strand
saat proses replikasi DNA. Selanjutnya akan dihubungkan oleh enzim
ligase sehingga untai menjadi DNA kontinyu.
Direction of Replication
 As the DNA unwinds, the template strand that is exposed in the 3:5 direction allows
the new strand to be synthesized continuously, in the 5:3 direction. This new strand,
which undergoes continuous replication, is called the leading strand.

DNA synthesis must start anew at the replication fork and proceed in the direction
opposite that of the movement of the fork until it runs into the previously replicated
segment of DNA. This process is repeated again and again, so synthesis of this strand
is in short, discontinuous bursts.

The newly made strand that undergoes discontinuous replication is called the
lagging strand. The short lengths of DNA produced by discontinuous replication of
the lagging strand are called Okazaki fragments, after Reiji Okazaki, who discovered
them.
 In bacterial cells, each Okazaki fragment ranges in length from about
1000 to 2000 nucleotides
in eukaryotic cells, they are about 100 to 200 nucleotides long.
Okazaki fragments on the lagging strand are linked together to
create a continuous new DNA molecule.
MUTASI GEN
Mutation and DNA
Mutation = change(s) in the nucleotide/base sequence of DNA; may occur
due to errors in DNA replication or due to the impacts of chemicals or
radiation to the DNA molecule

Mutation may result in coding sequences for new amino acids in proteins or
not!
Mutation(Permanent, heritable DNA changes)

Point mutation (base substitutions)

Missense mutation
Nonsense mutation (premature stop)
Silent mutation

Insertions/deletions
Frameshift mutation
1. Point mutations affect single sites on DNA
Substitution of 1 base for another
Deletion/addition of a single base
Deletion/addition of a small number of bases

If purine (A/G) or pyrimidine (T/C) substitutes for itself = transition substitution

If purine substitutes for pyrimidine or vice versa = transversion substitution
Results of point mutations

Silent mutations = due to redundancy of the Genetic

Code, most point mutations are silent do not code
for a different amino acid

Missense mutations = produces change in amino acid

in protein but does not change the function of the
protein

Nonsense mutations = produces a STOP codon in the

midst of the mRNA transcript; can produce a non-
functional protein
Sample outcome of DNA code

Methionine, proline, threonine, arginine, stop

Silent mutation

Due to redundancy of Genetic Code, no change in amino acid sequence

is produced!!
Missense mutation

Missense mutation produces a change in amino acid sequence in protein product

(Histidine in for Arginine); may change function of protein or may not!

Contoh lain: sickle cell disease GAG (Glu) GTG (Val)

Nonsense mutation

Bad news! nonsense mutation produces a STOP codon within the mRNA transcript leading
to a truncated protein. How short the protein product depends on where the STOP codon
was produced within the mRNA transcript.

Contoh lain: cystic fibrosis Glu (GAG) STOP (TAG/UAG)

 2. Chromosomal mutations change the
structure of whole chromosomes
Chromosomal mutations are more extensive, altering the entire
chromosomal structure

These kinds of mutations occur through:

Deletions
Duplications

Inversions
Translocations

Deletions to chromosome

If too much information is lost, it may be fatal

to the organism and may result in early death
(e.g., Cri-du-chat syndrome large deletion
from chromosome #5)
Duplications within chromosome

Effect of base duplications depend on location within the

chromosome whether or not duplication resides in coding or
non-coding region of DNA
Inversions within chromosome
Translocations within chromosome

Can be caused due to abnormal synapsis event at Meiosis I by

incorrect chromosomes coming together.
Associated with 2 forms of leukemia oncogenes translocated
to incorrect regions within chromosomes of leukocytes (white
blood cells)
Translokasi kromosom menghasilkan protein bcr-abl pada leukimia
Cause of Mutation (mutagen)

Spontaneous mutation: Occurs in DNA replication

Chemical mutagens
Base pair changers (nitrous acid)
Base analogues (e.g.. 5 bromouracil)
Frameshift mutagens (aflatoxin, benz-(a)-pyrene)
Radiation
X rays, gamma rays break DNA, bases
UV light causes knots in DNA strand
Mutation is important for evolution

If no changes to genomes occur over time, there

would be no evolution
Too much change in the DNA is harmful
Too little does nothing
A balance exists between the amount of new variation and
the overall health (adaptiveness) of the new variant
individual

Differences between closely related organisms show

closely matched DNA sequences that diverged at
some past time and that was adaptive for a given
environment
Our understanding of genes changes with new
information
G. Mendel followed traits caused by single gene pairs
T.H.Morgan mapped locations of genes on chromosomes
Jacob and Monod modelled 1 gene, 1 polypeptide
Later, it was found some DNA coded for only RNA (e.g., rRNA and tRNA)

Some DNA expresses regulatory genes in some people and the same DNA may not
express in others

To understand the nature of genes, we must consider their molecular structure

(genotype) as well as their phenotypic expression