GEN &

EKSPRESI GEN

PURNOMO SURYOHUDOYO

DEPT. OF BIOCHEMISTRY

SCHOOL OF MEDICINE

AIRLANGGA UNIVERSITY

TONGGAK SEJARAH PERKEMBANGAN BIOLOGI MOLEKULER

1865 : Mendel : Faktor pewarisan 1903 : Sutton : Penurunan sifat ada hubungan dengan pembelahan kromosom 1906 : Johansen : Istilah Gen 1910 -1915 : Morgan dkk. : Gen terdapat dalam kromosom 1942 : Avery, Mc Leod, Mc Carthy : Gen = DNA1953* : Watson &Crick : Struktur Heliks Kembar DNA 1958 : Crick : Dogma Sentral : DNA RNA Protein

* = Awal Kelahiran Biologi Molekuler

GENE DATE LINE1865 Mendel : Hereditary factor later ( 1906) called “gene”

by Johansen

1910 – 1915 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’

Ikatan3’, 5’ fosfodiester

Ujung 3’

DNADNA RNARNA

Ujung 5’ 3’ 5’ 3’Ujung 5’ 3’ 5’ 3’ 3’ 5’3’ 5’

Rantai Kembar TunggalRantai Kembar Tunggal

Gula Deoksiribosa RibosaGula Deoksiribosa Ribosa (dR) (R)(dR) (R)

Basa G C G CBasa G C G C

T A U AT A U A

Perbedaan DNA - RNAPerbedaan DNA - RNARantai : Kembar – TunggalRantai : Kembar – TunggalGula : Deoksiribosa – RibosaGula : Deoksiribosa – RibosaBasa : T - U Basa : T - U

T: Timin, U : Urasil, T: Timin, U : Urasil,

G : Guanin G : Guanin A : Adenin A : Adenin C : Sitosin C : Sitosin

STRUKTUR NUKLEOTIDA

BESERTA KOMPONEN

PEMBENTUKNYA

Atom N

G Guamin C Sitosin

A Adenin T Timin

dR Deoksiribosa

ikatan hidrogen

PASANGAN WATSON - CRICK

DNA

Rantai ganda

Anti-paralel

Komplementer

Atom NdR Deoksiribosa

REPLIKASI SEMIKONSERVATIF DNA

A

A

T

T

G

G

C

C

BERKAS BARU

BERKAS LAMA

A

A

T

T

G

G

C

C

A

A

T

T

G

G

C

C

CHROMOSOMES

CHROMOSOMES

AUTOSOMAL : 1 22

SEX : X, Y

SOMATIC CELLS (2n)

♂ : 44, XY

♀ : 44, XX

GERM CELLS (n)

OVA : 22, X

SPERMS : 22,X or 22,Y

BENTUK KROMOSOM

TELOSENTRIK

SentromerSentromer

METASENTRIK Kromosom 1, 3, 19, 20

(Lengan Pendek)

SUBMETASENTRIK(Lengan Panjang)

AKROSENTRIK Kromosom 13, 14, 15, y)

Satelit Penyempitan kedua (C2NO Construction)

tak ada pada manusia TELOSENTRIK

PEWARNAAN KROMOSOM

PEWARNAAN KROMOSOM

PEWARNAAN GIEMSA

KROMOSOM 13

1990. BM 1.4

ORGANISATION OF DNA WITHIN CHROMOSOMES ( 1 )

NUCLEOSOMESPACERDNA (200 KB)

APPEARANCE OF EXTENDED 10 NmCHROMATIN FIBRE IN E.M

10 nM

STRUCTURE OF NUCLEOSOME

STRUCTURE OF 30 nM FIBRE

HISTONE OCTOMER2 H2 A2 H2 B2 H3

2 H4

DNADOUBLE HELIX

NUCLEOSOMESPACER

DNA

ORGANISATION OF DNA WITHIN CHROMOSOMES ( 2 )

1990. BM 1.5

30 nM fiber

DNA loops

DNA back bone

700 nM

NON – HISTONEPROTEIN SCAFFOLD

DNA Loops

Coiled DNA back bone

1. Coiled DNA

backbone

2. DNA loops

3. Protein scaffold

4. Connecting scaffold

at Centro mere

EKSPRESI GEN

DNA

RNA

(m RNA) (r RNA) (t RNA)

PROTEIN = POLIPEPTIDA

POLINUKLEOTIDA

MELIBATKAN :

MEKANISME KERJA GEN

1909: GARRODMengemukakan 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 AminoAsam AminoStruktur Umum : NHStruktur Umum : NH22 CH C CH C

RR OHOH

OO

STRUKTUR UMUM PROTEIN (POLIPEPTIDA)STRUKTUR UMUM PROTEIN (POLIPEPTIDA)

Ujung N Ujung C

Ikatan Peptida

NH2 CH C NH CH C NH CH C

R1RnR 2

O

OH

OO

Asam AminoAsam AminoStruktur Umum : NHStruktur Umum : NH22 CH C CH C

RR OHOH

OO

Alanin ALA AArginin ARG RAspartat ASP DAsparagin ASN N

Fenilalanin PHE FGlisin GLY GGlutamat GLU EGlutamin GLN Q

Histidin HIS HIsoleusin ILE ILeusin LEU LLisin LYS K

Melionin MET MProlin PRO PSerin SER SSistein CYS C

Treonin THR TTirosin TYR YTriptofan TRP WValin VAL V

Jenis Asam Amino

H

H

H

H

H

H

H

H

OH

O

C C

C C

N

C PROLIN

CENTRAL DOGMA

DNA RNA PROTEIN

GENE PROTEIN

CELL PHENOTYPE

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

CENTRAL DOGMA

TRANSCRIPTION

Sense A T T G C C T G C

Anti – Sense T A A C G G A C GTemplate

A U U G C C U G C 5’

5’

5’ 3’

5’

3’

3’

5’

3’

Direction of Transcription

TRANSLATION

A U U G C C U G C

ILE ALA CYS C

Direction of TranslationN C

N

(m) RNA

PROTEIN

DNA

RNA

DIRECTION OF GENE TRANSCRIPTION

DIRECTION OF TRANSCRIPTION

DIRECTION OF TRANSCRIPTION

5’ 3’

5’3’DNA

GENE YGENE X

SANDI ASAM AMINO

I : Sandi Inisiasi ; Term : Sandi Terminasi

UUU UUU phephe

UUC UUC phe phe

UUA UUA leuleu

UUG UUG leuleu

UCU UCU serser

UCC UCC serser

UCA UCA serser

UCG UCG serser

UAU UAU tyrtyr

UAC UAC tyrtyr

UAA UAA termterm

UAG UAG termterm

UGU UGU cyscys

UGC UGC cyscys

UGA UGA termterm

UGG UGG trptrp

CUU CUU leuleu

CUC CUC leuleu

CUA CUA leuleu

CUG CUG leuleu

CCU CCU propro

CCC CCC propro

CCA CCA propro

CCG CCG propro

CAU CAU hishis

CAC CAC hishis

CAA CAA glngln

CAG CAG glngln

CGU CGU argarg

CGC CGC argarg

CGA CGA argarg

CCG CCG argarg

AUU AUU ileile

AUC AUC ileile

AUA AUA ileile

AUG met (I)AUG met (I)

ACU ACU thrthr

ACC ACC thrthr

ACA ACA thrthr

ACG ACG thrthr

AAU AAU asnasn

AAC AAC asnasn

AAA AAA lyslys

AAG AAG lyslys

AGU AGU serser

AGC AGC serser

AGA AGA argarg

AGG AGG argarg

GUU GUU val val

GUC GUC valval

GUA GUA valval

GUG val (I)GUG val (I)

GCU GCU ala ala

GCC GCC alaala

GCA GCA alaala

GCG GCG alaala

GAU GAU aspasp

GAC GAC aspasp

GAA GAA gluglu

GAG GAG gluglu

GGU GGU glygly

GGC GGC glygly

GGA GGA glygly

GGG GGG glygly

SANDI GENETIK1. 3 Nukleotida menyandi 1 asam amino (Sandi Triplet)1. 3 Nukleotida menyandi 1 asam amino (Sandi Triplet)

2. 1 asam amino dapat disandi oleh lebih dari 1 triplet nukleotida 2. 1 asam amino dapat disandi oleh lebih dari 1 triplet nukleotida

(Sandi Degeneratif)(Sandi Degeneratif)

6 sandi : leu, arg, ser6 sandi : leu, arg, ser

4 sandi : val, pro, thr, gly4 sandi : val, pro, thr, gly

3 sandi : ile3 sandi : ile

2 sandi : hyr, his, gln, asn, Lys, asp, glu, cys2 sandi : hyr, his, gln, asn, Lys, asp, glu, cys

1 sandi : met, trp 1 sandi : met, trp

4. 4. UAA, UAG, UGAUAA, UAG, UGA : merupakan tanda pengakhiran sintesis protein : merupakan tanda pengakhiran sintesis protein Sandi TerminasiSandi Terminasi

3 sandi ini tidak sesuai dengan asam amino manapun 3 sandi ini tidak sesuai dengan asam amino manapun Sandi tanpa Sandi tanpa anti (Nonsense Code)anti (Nonsense Code)

3. 3. AUG, GUG merupakan tanda awal sintesis protein. AUG, GUG merupakan tanda awal sintesis protein. Sandi InisiasiSandi Inisiasi

(AUG (AUG met, GUG met, GUG val) val)

5. Sandi Genetik (hampir) Universal5. Sandi Genetik (hampir) Universal

(ada beberapa perkecualian khususnya pada MT-DNA)(ada beberapa perkecualian khususnya pada MT-DNA)

V I R I O N S 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 : RhabdovirusClass 3 : Double stranded RNA

Example : ReovirusClass 4 : Single Stranded RNA (+) but Replicate Through DNA Intermediate Retrovirus

Example : HIV

CLASS 1

CLASS 2 (+) DNA(+) DNA as template

(-) DNA(-) DNA as template

(+)(+)(+)

DNA

Virions Example : Parvovirus

(+)(-)

Replication

(+)(-)

(+)

Virions Example : Adeno Virus

DNA

(-)

CLASS 3 :

Example : HBV (Hepatitis B Virus)(+) RNA act as mRNA

(-) (-) DNA as template

(+)(-)

DNA

Virions

(+)

(-) DNA as template

(+)

(+)

DNA

Double Stranded DNA

DNA

(+) RNA as template

DNA

RNA(+)

CLASS 1

CLASS 2

(+) : RNA(+) : RNA as template

(-) : RNA(-) : RNA as template

(+)(+)(+)

RNA

Virions Example : Picorna Virus

(-) : RNA(-) : RNA as template

(-) : RNA(+) : RNA as template

(-)(-)(-)

RNA

Virions Example : Rhabdovirus

CLASS 3

CLASS 4

(+) : RNAReverse transcription

(-) : DNA(+) : DNA as template

(+)(+)

RNA

Virions Example : Retroviruses

(+)

Replication(-)

(+)(-)(+)

RNA

Virions Example : Reovirus

RNA

(-)

(+)

Integrate into host genome (-) DNA as template

(+)(-)Double stranded DNA

(e.g. Hiv