GENE TRANSCRIPTION

DNA RNA
RNA: always single stranded
Only 1 strand of DNA act as a template for
transcription
RNA synthesis: 5 3
Steps: Initiation, Elongation, Termination
Required components:
- Template ssDNA
- RNA polymerase
- Promoter element
- Upstream DNA sequences, Enhancer element
- Transcription factors, activator

GENE TRANSCRIPTION IN
PROKARYOTES
1 type of RNA polymerase in E.coli:
holoenzyme
(core enzyme+ subunit)
Core enzyme:
2
,

, , () subunits
subunit: Initiation factor
Promoter: 10 dan 35 region

GENE TRANSCRIPTION IN
PROKARYOTES
Separation of double helix DNA (17 base
pairs: ~2 helix turns) Open promoter
complex
Synthesis of RNA chain starts with pppA
or pppG
subunit dissociates after addition of the
10
th
nucleotide (position +10)
Elongation: NusA protein
DNA-RNA hybrid: ~ 12 nucleotides
Termination: (rho) factor, oligo (dA-rU)







GENE TRANSCRIPTION
IN PROKARYOTES
Elongation: NusA protein
DNA-RNA hybrid: ~12 nucleotides
Separated double helix DNA ~17
nucleotides
Termination: - (rho) factor dependent
- factor independent

Upstream DNA sequences
Activator and Repressor

GENE TRANSCRIPTION
IN EUKARYOTES
RNA polymerase enzyme: monomer
3 types: RNA pol. I, II, III
RNA pol I: 18S, 5,8S dan 28S rRNA
RNA pol II: mRNA dan UsnRNA except
U6snRNA
RNA pol III: tRNA, 5S rRNA, U6snRNA
Transcription factor
Enhancer element
Transcription Factor
Protein
2 types:
- DNA binding protein
-non DNA binding protein
DNA binding protein has 2 domains:
- DNA binding domain
- Transcription activation domain

TRANSCRIPTION FACTOR
Stimulation of Gene Transcription:
helps RNA polymerase binding to
promoter or
affects reaction rate of transcription
initiation catalyzed by RNA
polymerase which bound to
promoter
TRANSCRIPTION FACTOR
Identification of DNA-binding protein:
- Mobility-shift assay
- DNase-footprinting assay
4 structural types of transcription factor:
- Helix-turn-helix (Homeodomain)
- Zincfinger
- Leucine zipper
- Helix-loop-helix

ENHANCER ELEMENT
Far from start point of transcription,
at 5 region (upstream)
Does not act as a promoter, but
enhances the promoter function by
interaction with DNA binding protein
Only active in the specific cell type
Regulated by several inducing
signals