SINTESIS PROTEIN PADA EUKARIOT

Evi Umayah Ulfa

Three eukaryotic polymerases

Type Location Substrate a-amanitin

RNA Pol I
RNA Pol II

Nucleoli
Nucleoplasm Nucleoplasm

Most rRNAs gene

All protein-coding genes and some snRNA genes tRNAs, 5S rRNA, U6 snRNA and other small RNAs

Insensitive
Very sensitive Moderately sensitive

RNA Pol III

PROMOTER
Promoters usually have 4 Elements: 1. TATA Box (25) 2. Initiator Element (Inr) 3. Downstream Core Promoter Element 4. CAAT box and some contain GC Box (-40 s/d -150)

TRANSCRIPTION INITIATION COMPLEX FORMATION

5 CAP

From GTP

Initiating nucleotide

The structure of the 5cap of eukaryotic mRNAs.

The formation of the 5cap of eukaryotic mRNAs.

SPLICING

 Mechanisms Transesterification

TRANSLASI PADA EUKARIOT

Translational control
Structure of eukaryotic and prokaryotic mRNAs:

Faktor-faktor yang Terlibat dalam Translasi

Inisiasi :
Kodon inisiasi : AUG Aminoasil tRNA : Metionin Eukaryot initiation factors (eIF) : eIF 1-6

Pemanjangan
Elongation Factor (EF)-Tu Enzim peptidil transferase EF-G

Terminasi
Kodon terminasi (UAA, UGA, UAG) eRF 1 dan 3

Model of eukaryotic ribosome

rRNAs are believed to play a catalytic role in protein synthesis. After removal of 95% of the ribosomal proteins, the 60S subunit can catalyze formation of peptide bonds. Ribosomal proteins are now believed to help fold the rRNAs properly and to position the tRNAs.

5 end (cap) dependent initiation:

The first step is the recognition of the 5 cap by eIF4F, which consists of three proteins, eIF4E, eIF4G and eIF4A.

Cap binding protein, eIF4E, binds to cap

The N-terminus of eIF4G binds eIF4E and the C-terminus binds eIF4A The 40S subunit binds to eIF4G via eIF3

Cap-Dependent Initiation of Protein Synthesis in Eukaryotes

An initiation complex forms at the cap with the 40S ribosomal subunit and other translation initiation factors.
The 40S complex then scans down the 5 untranslated region to the first AUG codon. A GTP hydrolysis step by eIF5 triggers GDP binding of eIF2 and release of initiation proteins. The 60S subunit joins the complex and the 80S ribosome initiates translate the ORF.

Elongation
Ribosome selects aminoacylated tRNA

eEF1a GTP

eEF1a and GTP are bound to aminoacylated tRNA

Ribosome catalyzes formation of a peptide bond Translocation is dependent on eEF2 and GTP hydrolysis Many ribosomes may translate mRNAs simultaneously on the same strand.
eEF2 GTP

P A

Termination Translation is terminated at one of three stop codons (UAA, UAG & UGA).

Termination codon at the A site is recognized by the release factor instead of a tRNA
The release factor binds the termination codon The peptide chain is then released followed by dissociation of the tRNA and the ribosome