The Central Dogma of Molecular Biology replication transcription translation.
Post on 14-Dec-2015
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Why RNA?
• Not all genes need to be turned on at once. –We can make an RNA transcript of just
ONE GENE– Now we can make the right protein at
the right time in the right location
Why RNA?
In EUKARYOTES…
• DNA cannot leave the nucleus• BUT proteins are built by the
ribosomes in the cytosol!• We need a messenger to transfer the
genetic code to the ribosomes
Initiation• RNA polymerase (RNAP) binds to the
double stranded DNA molecule at a promoter sequence (with the help of initiation factors)
• It is able to locally unzip DNA with its own built in helicase activity as it constructs an RNA transcript of the DNA
RNA Polymerase II
Promoters• DNA sequence
upstream of the gene being transcribed
• Determines where RNAP binds and where transcription begins
• Usually rich in Thymine and Adenine (“TATA” box)
Elongation
• One strand of the unzipped DNA acts as a template for RNA synthesis
SG
P SA
P ST
P SC
P SG
P SG
P SA
P SC
P SC
P3´ 5´
SG
PSG
PST
PSC
PSC
PSG
PSA
PST
PSC
P5´ 3´
Template Strand
Elongation
SU
P
SG
PSG
PST
PSC
PSC
PSG
PSA
PST
PSC
P5´ 3´
Template StrandSC
PSG
P SA
P ST
P SC
P SG
P SG
P SA
P SC
P SC
P3´ 5´
SA
PSG
PSC
PSC
PSU
PSG
PSG
P
5´ 3´
Coding Strand
Elongation
• mRNA is transcribed in the 5' to 3' direction
• DNA unwinds only in the region of transcription
• After transcription DNA recoils
• Several RNAPs can work on a single gene at once
Termination• A terminator sequence on the coding
strand tells RNAP when to stop transcribing the mRNA
• RNAP is released and reused and mRNA is released
Processing
• In Eukaryotic cells the RNA transcript is called pre-mRNA (or primary RNA) because it must still be modified before it leaves the nucleus
• Why processing?– Remove introns– Protects from degradation in the
cytoplasm
Splicing
• The Spliceosome is a complex of multiple proteins (snRNPs) and small nuclear RNA (snRNA)
• snRNA binds a specific mRNA sequences at the beginning and end of an intron forming a loop
• The loop is removed and exons are linked
Cap and Tail
S
G
PS
G
PS
U
PS
C
PS
C
PS
G
PS
A
PS
U
PS
C
P
5´ 3´
S
G
PS
G
PS
G
PS
G
P S
A
PS
A
PS
A
PS
A
P S
A
PS
A
PS
A
PS
A
P
cap poly A tail
mRNA transcrip
t
Cap and Tail
• To protect RNA from restriction endonucleases in the cytosol, a poly-A tail is added to the 3‘ end of the pre-mRNA
• As an attachment site for the ribosome, a 5‘ cap of modified Gs is added
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