Download (+) DNA

GEN
&
EKSPRESI GEN
PURNOMO SURYOHUDOYO
DEPT. OF BIOCHEMISTRY
SCHOOL OF MEDICINE
AIRLANGGA UNIVERSITY
TONGGAK SEJARAH
PERKEMBANGAN BIOLOGI MOLEKULER
1865
1903
1906
1910 -1915
1942
1953*
1958
: Mendel
: Sutton
: Faktor pewarisan
: Penurunan sifat ada hubungan dengan
pembelahan kromosom
: Johansen
: Istilah Gen
: Morgan dkk. : Gen terdapat dalam kromosom
: Avery, Mc Leod, Mc Carthy
: Gen = DNA
: Watson &Crick : Struktur Heliks Kembar DNA
: Crick
: Dogma Sentral : DNA  RNA  Protein
* = Awal Kelahiran Biologi Molekuler
GENE DATE LINE
1865
1910 – 1915
Mendel : Hereditary factor later ( 1906) called “gene”
by Johansen
Morgan : Genes resides in chromosomes
1942
Avery, Mc Leod & Mc. Carthy
Gene = DNA
1953
Watson & Crick : Structure of DNA :
- Double Helix
- Anti Paralel
Strands
- Complementary
1958
Crick proposed the central Dogma :
DNA  RNA  Protein
A theory later confirmed by the discovery of mRNA by
Brenner & Jacob (1961)
STRUKTUR POLINUKLEOTIDA
Ujung 5’
Ikatan
3’, 5’ fosfodiester
Ujung 3’
DNA
RNA
Ujung
5’ 3’
3’ 5’
5’
Rantai
Kembar
Tunggal
Gula
Basa
Deoksiribosa
(dR)
3’
Ribosa
(R)
G
C
G
C
T
A
U
A
Perbedaan DNA - RNA
Rantai : Kembar – Tunggal
Gula : Deoksiribosa – Ribosa
Basa : T - U
T: Timin, U : Urasil,
G : Guanin
A : Adenin
C : Sitosin
STRUKTUR NUKLEOTIDA
BESERTA KOMPONEN
PEMBENTUKNYA
PASANGAN WATSON - CRICK
G
A
Atom N
Guamin
Adenin
C Sitosin
T Timin
dR Deoksiribosa
ikatan hidrogen
DNA
Rantai ganda
Anti-paralel
Komplementer

Atom N
dR Deoksiribosa
REPLIKASI SEMIKONSERVATIF DNA
A
T
G
C
T
A
C
G
A
T
A
T
G
C
G
C
T
A
T
A
C
G
C
G
BERKAS BARU
BERKAS LAMA
CHROMOSOMES
CHROMOSOMES
AUTOSOMAL
SEX
: 1  22
: X, Y
SOMATIC CELLS (2n)
♂
♀
: 44, XY
: 44, XX
GERM CELLS (n)
OVA
: 22, X
SPERMS
: 22,X or 22,Y
BENTUK KROMOSOM
METASENTRIK
Sentromer
Sentromer
Kromosom 1, 3, 19, 20
(Lengan Pendek)
SUBMETASENTRIK
(Lengan Panjang)
Satelit
Penyempitan kedua (C2NO Construction)
AKROSENTRIK
Kromosom 13, 14, 15, y)
TELOSENTRIK
TELOSENTRIK
tak ada pada manusia
PEWARNAAN
KROMOSOM
PEWARNAAN
KROMOSOM
PEWARNAAN GIEMSA
KROMOSOM 13
ORGANISATION OF DNA WITHIN CHROMOSOMES ( 1 )
APPEARANCE OF EXTENDED 10 Nm
CHROMATIN FIBRE IN E.M
NUCLEOSOME
SPACER
DNA (200 KB)
HISTONE OCTOMER
2 H2 A
2 H2 B
2 H3
2 H4
10 nM
STRUCTURE OF
NUCLEOSOME
DNA
DOUBLE HELIX
NUCLEOSOME
STRUCTURE OF
30 nM FIBRE
SPACER DNA
1990. BM 1.4
ORGANISATION OF DNA WITHIN CHROMOSOMES ( 2 )
30 nM fiber
DNA loops
DNA back bone
DNA Loops
Coiled DNA back bone
700 nM
NON – HISTONE
PROTEIN
SCAFFOLD
1. Coiled DNA backbone
2. DNA loops
3. Protein scaffold
4. Connecting scaffold
at Centro mere
1990. BM 1.5
EKSPRESI GEN
MELIBATKAN :
DNA
POLINUKLEOTIDA
RNA
(m RNA)
(r RNA)
(t RNA)
PROTEIN
=
POLIPEPTIDA
MEKANISME KERJA GEN
1909: GARROD
Mengemukakan pendapat bahwa penyakit alkaptonuria, suatu penyakit herediter
(keturunan) disebabkan oleh ketiadaan suatu enzim
1921: MULLER
Menemukan bahwa penyinaran dengan sinar – X menyebabkan mutasi
1942: BEADLE & TATUM
Penyinaran sinar – X terhadap ragi Neurospora crassa menyebabkan hilangnya
enzim-enzim tertentu
Gen  Enzim
1949: PAULING
Penyakit sel sabit (Sickle cell disease) suatu penyakit herediter, disebabkan oleh
adanya suatu Hb abnormal : Hb S
Gen  Protein
1958 : CRICK
Mengemukakan teori bahwa struktur DNA menentukan struktur RNA dan struktur
RNA menentukan struktur protein
1961: BRENNER & JACOBS
Menemukan mRNA
DOGMA SENTRAL
Urutan Nukleotida DNA  Urutan nukleotida mRNA  urutan As. Amino Protein
Asam Amino
Struktur Umum :
O
NH2
CH
R
O
NH2
CH
C
C
OH
O
NH
CH
C
O
NH
R2
R1
CH
Rn
Ikatan Peptida
Ujung N
STRUKTUR UMUM PROTEIN (POLIPEPTIDA)
Ujung C
C
OH
O
Asam Amino
Struktur Umum :
NH2
CH
C
OH
R
Jenis Asam Amino
Alanin
Arginin
Aspartat
Asparagin
ALA
ARG
ASP
ASN
A
R
D
N
Fenilalanin
Glisin
Glutamat
Glutamin
PHE
GLY
GLU
GLN
F
G
E
Q
Histidin
Isoleusin
Leusin
Lisin
HIS
ILE
LEU
LYS
H
I
L
K
Melionin
Prolin
Serin
Sistein
MET
PRO
SER
CYS
M
P
S
C
Treonin
Tirosin
Triptofan
Valin
THR
TYR
TRP
VAL
T
Y
W
V
H
H
H
H
C
H
C
H
C
C
N
H
H
C
O
OH
PROLIN
CENTRAL DOGMA
DNA
GENE
RNA
PROTEIN
PROTEIN
CELL PHENOTYPE
CENTRAL DOGMA
REPLICATION :
Old Strand
5’
A
T
T
G
C
C
T
G
C
3’
New Strand
3’
T
A
A
C
G
G
A
C
G
5’
3’
5’
Direction of Replication
TRANSCRIPTION
DNA
Sense 
5’
A
T
Anti – Sense 
T
3’
Template
RNA
5’
A
T
G
C
C
T
A
A
C
G
G
A
U
U
G
C
C
U
G
C
C
G
G
3’
5’
C
3’
5’
3’
Direction of Transcription
TRANSLATION
(m) RNA
5’
PROTEIN
N
A
U
ILE
N
U
G
C
ALA
C
Direction of Translation
C
U
G
CYS
C
C
DIRECTION OF
GENE TRANSCRIPTION
DIRECTION OF
TRANSCRIPTION
5’
3’
DNA
3’
GENE X
DIRECTION OF
TRANSCRIPTION
GENE Y
5’
SANDI ASAM AMINO
UUU
UUC
UUA
UUG
phe
phe
leu
leu
UCU
UCC
UCA
UCG
ser
ser
ser
ser
UAU
UAC
UAA
UAG
tyr
tyr
term
term
UGU
UGC
UGA
UGG
cys
cys
term
trp
CUU
CUC
CUA
CUG
leu
leu
leu
leu
CCU
CCC
CCA
CCG
pro
pro
pro
pro
CAU
CAC
CAA
CAG
his
his
gln
gln
CGU
CGC
CGA
CCG
arg
arg
arg
arg
AUU
AUC
AUA
AUG
ile
ile
ile
met (I)
ACU
ACC
ACA
ACG
thr
thr
thr
thr
AAU
AAC
AAA
AAG
asn
asn
lys
lys
AGU
AGC
AGA
AGG
ser
ser
arg
arg
GUU
GUC
GUA
GUG
val
val
val
val (I)
GCU
GCC
GCA
GCG
ala
ala
ala
ala
GAU
GAC
GAA
GAG
asp
asp
glu
glu
GGU
GGC
GGA
GGG
gly
gly
gly
gly
I : Sandi Inisiasi ;
Term : Sandi Terminasi
SANDI GENETIK
1. 3 Nukleotida menyandi 1 asam amino (Sandi Triplet)
2. 1 asam amino dapat disandi oleh lebih dari 1 triplet nukleotida
(Sandi Degeneratif)
6 sandi : leu, arg, ser
4 sandi : val, pro, thr, gly
3 sandi : ile
2 sandi : hyr, his, gln, asn, Lys, asp, glu, cys
1 sandi : met, trp
4. UAA, UAG, UGA : merupakan tanda pengakhiran sintesis protein
Sandi Terminasi
3 sandi ini tidak sesuai dengan asam amino manapun  Sandi tanpa
anti (Nonsense Code)
3. AUG, GUG merupakan tanda awal sintesis protein. Sandi Inisiasi
(AUG  met, GUG  val)
5. Sandi Genetik (hampir) Universal
(ada beberapa perkecualian khususnya pada MT-DNA)
VIRIONS
1. Viruses Cannot Replicate outside its host - cell
2. Outside its host cell, it forms Virions
3. A Virions consists of :
- Genetic Material (DNA or RNA)
- Nucleocapsid (Protein)  COAT
4. Some Virions have an extra coat called :
Envelope : Lipid bilayer originating from cell membrane
of host cell, studded with viral protein
5. Virusoids are viruses which cannot form virions without
the help of another virus called helper virus
Example : HDV (Hepatitis D Virus)
Helper Virus : HBV
6. VIROIDS are viruses (usually RNA) which never exist outside
their host cell
Example : certain plant viruses
CLASSIFICATION OF VIRUSES
DNA VIRUS
Class 1 : Doble Stranded DNA
Example : Adenovirus
Class 2 : Single Stranded DNA (+)
Example : Parvovirus
Class 3 : Double stranded DNA but replicate through RNA Intermediate
Example : Hepadna Virus
RNA VIRUS
Class 1 : Single Stranded RNA (+)
Example : Picorna Virus
Class 2 : Single Stranded RNA (-)
Example : Rhabdovirus
Class 3 : Double stranded RNA
Example : Reovirus
Class 4 : Single Stranded RNA (+) but Replicate Through DNA Intermediate
Retrovirus
Example : HIV
(+)
(-)
CLASS 1
DNA
Replication
(+)
(-)
(+)
(-)
Virions
CLASS 2
Example : Adeno Virus
(+) DNA
(+) DNA as template
(-) DNA
(-) DNA as template
(+)
(+) DNA
(+)
Virions
Example : Parvovirus
CLASS 3 :
(+)
(-) DNA
(-) DNA as template
(+) RNA
(+) RNA as template
(+) DNA
(-) DNA as template
(+) DNA
Double Stranded DNA
(+) DNA
(-)
Virions
Example : HBV (Hepatitis B Virus)
(+) RNA act as mRNA
CLASS 1
(+) : RNA
(+) : RNA as template
(-) : RNA
(-) : RNA as template
(+)
(+) RNA
(+)
Virions
CLASS 2
Example : Picorna Virus
(-) : RNA
(-) : RNA as template
(-) : RNA
(+) : RNA as template
(-)
(-) RNA
(-)
Virions
Example :
Rhabdovirus
CLASS 3
Replication
Virions
CLASS 4
(+)
(-)
RNA
(+)
(-)
(+)
(-)
RNA
Example : Reovirus
(+) : RNA
Reverse transcription
(-) : DNA
(+) : DNA as template
(+)
(-) Double stranded DNA
Integrate into host genome
(-) DNA as template
(+) RNA
(+)
(+)
Virions
Example : Retroviruses
(e.g. Hiv