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MUTASI GEN

Oktavia Rahayu A, S.Farm., M.Biomed - Program Studi Sarjana Farmasi 2017


Outline

Implikasi positif
Klasifikasi jenis dan negatif
Prinsip mutasi mutasi yang terjadinya
dapat terjadi mutasi dalam
bidang farmasi
Mutasi
• MUTASI  perubahan permanen sekuens nukleotida pada suatu molekul DNA
• Mutasi terjadi pada level gene
• Perubahan DNA
• Perubahan mRNA
• Kemungkinan perubahan protein
• Kemungkinan perubahan karakter
• Mutasi dapat diklasifikasikan berdasarkan
• Penyebab
• Mekanisme
Efek Mutasi
Positif
• Sejumlah kecil mutasi dapat meningkatkan fungsi protein tertentu 
terjadi dalam proses evolusi

Negatif
• Jika asam amino yang berubah merupakan asam amino pada sisi aktif
protein  mengganggu atau menekan fungsi protein
• Perubahan pada bagian tertentu protein dapat mengubah spesifisitas
protein  tidak mengenali target
Mutations occasionally produce a polypeptide with
an enhanced ability to function  May result in an
organism with a greater likelihood to survive and
reproduce
Start codon?
Stop codon? GENETIC CODE
TYPE OF MUTATION Silent mutation

Point mutation: base Mis-sense mutation


substitution Non-sense mutation
Sense mutation

Mutation
Point mutation: Insertion
Type of Frameshifts and
Reversions Deletion
Splicing mutations
mRNA Problems Polyadenylation site mutations
Trinucleotide Repeats
Gene rearrangement
Other mutation Gene duplication
Gene deletion
Point mutation: base substitution

Silent mutation
5’ AUG UUU ACA AAA CUG UAA 3’
met- phe- thr- lys- leu- COOH

5’ AUG UUU ACC AAA CUG UAA 3’


met- phe- thr- lys- leu- COOH
Mis-sense mutation
5’ AUG UUU ACA AAA UAA 3’
met-phe-thr- lys-COOH

5’ AUG UUU AUA AAA UAA 3’


met-phe-Ile- lys-COOH
Example: HbS, sickle cell hemoglobin, is a change in the beta-globin
gene, where a GAG codon is converted to GUG. GAG codes for
glutamic acid, which is a hydrophilic amino acid that carries a -1
charge, and GUG codes for valine, a hydrophobic amino acid. This
amino acid is on the surface of the globin molecule, exposed to water.
Under low oxygen conditions, valine’s affinity for hydrophobic
environments causes the hemoglobin to crystallize out of solution.
Non-sense mutation
5’ AUG UUU AUA AAA CUC UAA 3’
met- phe- Ile- lys- leu- COOH

5’ AUG UUU AUA UAA CUC UAA 3’


met- phe- Ile-COOH (stop codon)

Example: Hb-β McKees Rock. Normal beta-globin is 146 amino acids long.
In this mutation, codon 145 UAU (codes for tyrosine) is mutated to UAA
(stop). The final protein is thus 143 amino acids long. The clinical effect is
to cause overproduction of red blood cells, resulting in thick blood
subject to abnormal clotting and bleeding.
Sense mutation
Opposite of nonsense mutations  a stop codon is converted into an
amino acid codon. Since DNA outside of protein-coding regions
contains an average of 3 stop codons per 64, the translation process
usually stops after producing a slightly longer protein.

Example: Hb-α Constant Spring. alpha-globin is normally 141 amino acids


long. In this mutation, the stop codon UAA is converted to CAA
(glutamine). The resulting protein gains 31 additional amino acids
before it reaches the next stop codon. This results in thalassemia, a
severe form of anemia.
Frameshifts and Reversions
Frameshifts
Translation occurs codon by codon, examining nucleotides in groups
of 3. If a nucleotide or two is added or removed, the groupings of
the codons is altered. This is a “frameshift” mutation, where the
reading frame of the ribosome is altered.

Insertion Deletion
5’ AUG UUU CUC AUC ACC 3’ 5’ AUG UUU CUC AUC ACC 3’
met- phe- leu- Ile- thr met- phe- leu- Ile- thr

5’ AUG AUU UCU CAU CAC 3’ 5’ AUG UUC UCA UCA CC 3’


met- Ile- Ser- His- His met- phe- ser- ser-
Insertion/deletion?
example Hb-α Wayne. The final codons of the alpha globin chain are
usually AAA UAC CGU UAA, which code for lysine-tyrosine-arginine-
stop. In the mutant, one of the A’s in the first codon is deleted,
resulting in altered codons: AAU ACC GUU AAG, for asparagine-
threonine-valine-lysine. There are also 5 more new amino acids added
to this, until the next stop codon is reached.
Reversions
Reversion is second mutation that reverse the effects of an initial
mutation, bringing the phenotype back to wild type (or almost)
Frameshift mutations sometimes have “second site reversions”, where a
second frameshift downstream from the first frameshift reverses the effect.

Example: consider Hb Wayne above. If another mutation occurred that


added a G between the 2 C’s in the second codon, the resulting codons
would be: AAU ACG CGU UAA, or asparagine-threonine-arginine-stop.
Note that the last 2 codons are back to the original. Two amino acids are
still altered, but the main mutational effect has been reverted to wild type.
CYSTIC FIBROSIS
• Broken salt channel in cells
• strikes 1 in 2500 white births
• gene codes for a protein channel
that allows salt to flow across cell membrane
• broken protein doesn’t work as channel
• doesn’t allow salt out of cell, so water doesn’t flow out either
• thicker & stickier mucus coating around cells
• mucus build-ups in lungs & causes bacterial infections
• destroys lung function
• without treatment children die before 5; with treatment can live past their late 20s
Salt channel
EFFECT ON LUNGS transports salt through protein chann
out of cell
normal lungs airway Osmosis problems!
salt
normal mucus
H2 O salt channel
cells lining lungs

cystic fibrosis

salt


thick mucus
H2 O

mucus & bacteria build up


= lung infections & damage
mRNA Problems
Although many mutations affect the protein sequence directly, it is
possible to affect the protein without altering the codons
Splicing mutation
Intron removal requires several specific sequences. Most importantly,
introns are expected to start with GT and end in AG. Several beta
globin mutations alter one of these bases. The result is that one of the
2 introns is not spliced out of the mRNA. The polypeptide translated
from these mRNAs is very different from normal globin, resulting in
severe anemia.
Polyadenylation site mutations
The primary RNA transcript
of a gene is cleaved at the
poly-A addition site, and
100-200 A’s are added to the
3’ end of the RNA. If this site
is altered, an abnormally
long and unstable mRNA
results. Several beta globin
mutations alter this site: one
example is AATAAA ->
AACAAA. Moderate anemia
was the result.
Trinucleotide Repeats
• A fairly new type of mutation has been described, in which a
particular codon is repeated.
• During replication, DNA polymerase can “stutter” when it replicates
several tandem copies of a short sequence. For example,
CAGCAGCAGCAG, 4 copies of CAG, will occasionally be converted to 3
copies or 5 copies by DNA polymerase stuttering.
• Outside of genes, this effect produces useful genetic markers called
SSR (simple sequence repeats).
• Within a gene, this effect can cause certain amino acids to be
repeated many times within the protein. In some cases this causes
disease
• For example, Huntington’s disease is a neurological disease that generally
strikes in middle age, producing paranoia, uncontrolled limb movements,
psychosis, and death. Woody Guthrie, a folk singer from the 1930’s, had this
disease.
• The Huntington’s disease gene normally has between 11 and 33 copies of
CAG (codon for glutamine) in a row. The number occasionally changes.
People with HD have 37 or more copies, up to 200). The rate of copy
number change is much higher in HD people--too many copies makes the
repeated sequence more subject to DNA polymerase stuttering during
meiosis.
• Interestingly, the age of onset of the disease is related to the number of CAG
repeats present: the more repeats, the earlier the onset.
Types of Mutations according to Cause:

• Disebabkan oleh mutagen


Induced • Mutagen  senyawa kimia atau fisika
mutation yang menyebabkan perubahan sekuens
basa pada molekul DNA
• Penyebabnya tidak jelas
Spontaneous
mutation • Rate: 1 / 106 dalam suatu gen yang
bereplikasi
Penyebab mutasi spontan
• Abnormalitas pada saat crossing over
• Abnormalitas pemisahan kromosom saat meiosis
• Terjadi kesalahan oleh DNA polimerase saat
replikasi
• Perubahan DNA yang dipicu oleh senyawa kimia
hasil metabolisme endogen
• Integrasi elemen transposable

32
CAUSES OF INDUCED MUTATION
A. Deamination and methylation
• Contoh?
• heat
Physical B. Depurination
• ultraviolet
mutagens radiation • Contoh?

C. Alkylation of guanine
• Nitrosoguanine

Chemical • nitrous acid D. Base analog


• benzpyrenes
mutagens • 5 bromouracil, Acrydine orange
• aflatoxin
E. UV-light-induced thymine dimers
• base analogues
Factor induced mutation
A. Deamination and methylation
• Contoh?

B. Depurination
• Contoh?

C. Alkylation of guanine
• Nitrosoguanine

D. Base analog
• 5 bromouracil, Acrydine orange

E. UV-light-induced thymine dimers


• Ultraviolet radiation 
pembentukan dimer timin (atau
dimer Cytosines atau Thymine –
Cytosine, lebih jarang)
• Dimer  blok replikasi DNA
• Mekanisme repair DNA 
pemotongan daerah dimer dan
penyambungan kembali DNA
• Mutasi terjadi jika mekanisme
repair tidak berhasil
DNA REPAIR
A. Excition pair

B. Mismatch pair

C. Replication repair of UV-damaged DNA

D. Double strand repair by homologous


recombination
PENYEBAB MUTASI DAN MEKANISME PERBAIKANNYA
CHROMOSOME MUTATIONS

• May Involve:
• Changing the structure of
a chromosome
• The loss or gain of part of
a chromosome
TYPES OF MUTATIONS

Chromosome mutations

• Changes in chromosome structure

Genome mutations

• Changes in chromosome number

Single-gene mutations

• Relatively small changes in DNA


structure
• Occur within a particular gene

47
CHROMOSOME MUTATIONS
• Five types exist:
Deletion
Inversion
Translocation
Nondisjunction
Duplication
DELETION
• Terjadi pemotongan bagian kromosom sehingga
ada bagian yang hilang dari suatu kromosom
INVERSION
• Terjadi saat ada bagian kromosom yanga terpotong
kemudian menyambung kembali dengan
lokasi/urutan yang berkebalikan dari kondisi
asalnya
DUPLICATION
• Terjadi saat sebagian dari kromosom mengalami penggandaan
TRANSLOCATION
• Melibatkan dua
kromosom non
homolog
• Terjadi transfer suatu
bagian kromosom ke
kromosom lain
NONDISJUNCTION
• Terjadi saat ada kegagalan pemisahan
kromosom dalam meiosis
• Mengakibatkan sel gamet memiliki terlalu
sedikit/terlalu banyak kromosom
• Kelainan:
• Down Syndrome – three 21st chromosomes
• Turner Syndrome – single X chromosome
• Klinefelter’s Syndrome – XXY chromosomes
NON DIS-JUNCTION OF SEX CHROMOSOMES
TURNER’S SYNDROME

• Gamete with no sex chromosomes fuses with


normal X gamete
• Zygote has chromosome complement 2n = 45

• Individuals are female and short in stature

• Infertile because ovaries haven’t developed


normally
KLINEFELTER’S SYNDROME

• XX egg fertilised by normal Y sperm


or
• Normal X egg is fertilised by an XY sperm

• Zygote has chromosome complement 2n = 47 (44


+ XXY)
• Always male
• Normally
infertile
• Cannot
produce sperm