Transcription:

1. Initiation
a. RNA polymerase: an enzyme that catalyzes assembly of RNA nucleotides into
RNA strand for initiating transcription of gene
b. Promoter:
i. For prokaryotes: Site to which RNA polymerase binds
ii. For eukaryotes: Site to which general transcription factors bind and
recruit RNA polymerase for initiating transcription of gene
c. Promoter sequences: DNA sequences that define where transcription
of gene by RNA polymerase begins. 
d. TATA box
i. Usually the initiator sequence identified
ii. TATA box determines where transcription will initiate
iii. Type of promoter sequence
e. Transcription initiation complex: Combinations of TFs and RNA polymerase II
f. THE PROCESS
i. Transcription factors bind to promoter in an area within promotor called
TATA box, a special initiator sequence
ii. Other TFs bind and recruit RNA polymerase II
iii. RNA polymerase binds in an orientation that allows for the initiating
transcription to happen at correct place
iv. RNA polymerase II unzips or unwinds DNA
v. DNA unwound at front of RNA polymerase II to expose template strand
vi. Enzyme begins RNA synthesis at transcription start point
vii. Transcription factors released
2. Elongation
a. RNA polymerase II moves along DNA
b. Unwinds it
c. Adds new RNA molecules to transcript in 5’3’ direction
d. Behind enzyme, DNA strands reform into double helix
3. Termination
a. Complete RNA molecule released from template DNA
b. RNA polymerase II leaves DNA
c. DNA double helix reforms
d. Transcript of protein coding gene is pre-mRNA molecule
i. Processed to produce translatable mRNA
Translation Initiation:
1. Met-tRNA is initiator tRNA
a. Initiator tRNA has anticodon 3’-UAC-5’ complementary to AUG start codon
b. Initiator Met-tRNA + GTP binds to small ribosomal subunit on P site and forms
complex
2. Met-tRNA + GTP + small ribosomal subunit complex binds to 5’ cap of mRNA and moves
along mRNAPROCESS IS CALLED SCANNING
i. Until it reaches AUG start codon in P-site
 1. Base pairing occurs between codon and anticodon of initiator
Met-tRNA
3. Large ribosomal subunit binds and GTP is hydrolyzed, completing initiation
a. Ribosome is ready for next stage of translation, elongation
*each step aided by protein called initiation factor (Ifs released when initiation is complete in
step three)
Translation elongation
1. Aminoacyl-tRNA binds to codon in site A, GTP hydrolyzed here
a. Elongation factor facilitates binding and is bound to aminoacyl-tRNA
i. Elongation factor is released once tRNA binds to codon
2. Peptide bond is formed between C-terminal end of growing polypeptide on P site tRNA
and amino acid on A site tRNA
a. Peptadyl transferase cleaves amino acid (methionine, the initator) from tRNA in
P site, and forms peptide bond between the initiator and amino acid on the tRNA
in A site
b. Empty tRNA in P site, while the A site tRNA has two amino acids attached to it
i. The polypeptide chain grown on the A site
c. Peptidyl-tRNA: A tRNA with growing polypeptide chain of two or more amino
acids
3. Ribosome translocates to next codon along mRNA, while the tRNAs remain bound to
mRNA in their same positions (ribosome moves from 5’ to three prime ie left to right)
a. An elongation factor used for translocation, then released
b. Nrg from GDPGDP + P hydrolysis used
c. After translocation, tRNA that was in P site is now in E site
i. Peptadyl-tRNA that was in A site is now in the P site
4. Empty tRNA is released from E site
a. Ribosome is ready to begin next round of elongation cycle
*Elongation highly similar in eukaryotes and bacteria, no significant differences
Translation termination
1. Ribosome reaches a stop codon, UAG, UAA, or UGA in the A site
2. No more tRNAs have a anticodon that can pair with stop codon, so…
a. Release factor binds to stop codon in A site
i. Shape of release factor mimics that of tRNA
ii. Release factor: A protein that recognizes stop codons in A site of
ribosome translates an mRNA and terminates translation
3. Peptidyl transferase cleaves completed polypeptide chain from P site of the ribosome
a. Peptidyl transferase is stimulated by the release factor to cleave
4. Polypeptide released
5. Empty tRNA and release factor are released
a. Large and small ribosomal subunit separate and leave the mRNA