Transcription

and Translation

Chapter 14
 Central Dogma
• DNA mRNA Protein

• DNA DNA – Replication –

nucleus
• DNA mRNA –
Transcription – Nucleus
• mRNA Protein –
Translation – Ribosome in the
cytoplasm
Central Dogma
RNA vs. DNA
 DNA vs. RNA
DNA RNA
• Nucleus only • Nucleus and cytoplasm
• Double Stranded (ribosome)
• ATCG • Single Stranded
• Deoxyribose Sugar • AUCG
• Ribose sugar
• 3 main types
mRNA - messenger
tRNA - transfer
rRNA - ribosomal
 Triplet Code: Codons
Universal code: Strong evidence for evolution
3 Nucleotides = 1 amino acid Codon Chart pg. 273

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 Transcription
14_08TranscriptionIntro_A.swf

• DNA  mRNA
• Copying of DNA’s
message to mRNA
• Occurs in the nucleus
• Pre-mRNA is
processed into mRNA
and then leaves the
nucleus for the
cytoplasm (ribosome)
 Detailed Transcription
• Initiation: RNA polymerase binds to DNA at promoter
region.
– Promoter is before the gene that is to be transcribed
(eukaryotes need transcription factors to help RNA
polymerase bind to the TATA box)
– Determines which strand of DNA to use
• Elongation - adds nucleotides to mRNA strand based on
DNA strand in a 5’ 3’ direction (adding only to the
3’ end).
• Termination – RNA polymerase “falls off” the DNA
strand when the termination sequence (terminator) is
reached.
– AAUAAA in eukaryotes, this is now pre-mRNA
 Transcription
Transcription Initiation Eukaryotes Transcription Process
 RNA Processing
• Pre-mRNA RNA
• 5’ cap – guanine and phosphate cap on the 5’ end of
mRNA
• 3’ poly-A tail – 50 to 250 Adenines are added to
the 3’ end of the mRNA
• Both the 5’ cap and 3’ poly-A tail facilitate the
export of mRNA from the nucleus
• Both protect the mRNA from degradation by
hydrolytic enzymes in the cytoplasm
• Both help ribosomes attach to the 5’ ends of the
mRNA strand
RNA processing
 Alternative RNA splicing
• Introns – non-coding sections of mRNA
– Don’t leave the nucleus – only on pre-mRNA
• Exons – coding sections (expressed) of mRNA
– Exit the nucleus – final mRNA
• Keeps the exons gets rid of the introns
• snRNPs – cut the introns
– Ribozyme – RNA that functions like an enzyme
• Spliceosomes – join remaining exons together to
form final mRNA
 Translation
14_18TranslationIntro_A.swf

• mRNA  protein
• Process of mRNA
converting to a protein
• Occurs in the cytoplasm –
ribosome

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 tRNA
• Translator of mRNA’s message is tRNA –
transfer RNA
– 80 nucleotides long
– Hairpin shape – L shaped
• One end contains an anticodon which pairs
with the codon on the mRNA
– Codons determine which amino acid is coded for
by the DNA
• The other end contains an amino acid
attachment site
– Aminoacyl-tRNA synthetase attaches the correct
amino acid to the tRNA 15
tRNA

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 Ribosomes
• Pair codons on mRNA with anticodons on
tRNA to form polypeptides
• Made of large and small subunits
– rRNA – ribosomal RNA
– Made in the nucleolus
• Contain multiple binding sites
– mRNA binding site
– P site – peptidyl – tRNA site
– A site – aminoacyl – tRNA site
– E site – exit site

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Ribosomes

18
 Making a protein
• Initiation
– Small subunit binds to mRNA
– Start codon AUG – methionine at P site
• Elongation
– A site recognizes codon and pairs with correct tRNA
– Peptide bond forms between the carboxyl end of the polypeptide at the
P site and amino acid at the A site
– Amino acid in the A site translocates to the P site
• Termination
– Stop codon is reached at the A site
• UAA, UAG, UGA
– Release factors free the polypeptide from the ribosome

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 Making a Protein
ProteinSynthesis.mpg
TRANSCRIPTION DNA

mRNA
Ribosome
TRANSLATION
Polypeptide

Amino
Polypeptide acids

tRNA with
amino acid
Ribosome attached
Tr p
Phe Gly

tRNA
C
C
GC G
A
Anticodon
A A A
U G G U U U G G C

5 Codons 3
mRNA
Figure 17.13
 Proteins
• Fold spontaneously into primary,
secondary, and tertiary structures.
• Chaperone proteins assist in
folding.
• Some polypeptides become
quaternary with multiple subunits
• Signal peptide – directs proteins
through the endomembrane
system

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Targeting Polypeptides to locations

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 Prokaryotes vs. Eukaryotes
Eukaryotes
Prokaryotes • Locations:
• No nucleus – Free cytosol Ribosomes: Stay in the cell
somewhere (free floating, mitochondria, etc)
– Tanscription and translation – Rough ER Ribosomes: SRP sends ribosome to go
same location to ER, becomes protein in endomembrane
system, or cell membrane protein or is secreted
• Smaller ribosomes from cell (all in vesicles)
• Nucleus
– Pre-mRNA  mRNA
– transcription
• Larger ribosomes
– translation

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 Mutations
• Point Mutations – one base
altered
– Base-pair substitution
• Silent mutation – no
effect
• Missense mutation –
changes an amino acid
• Nonsense mutation –
creates a stop codon
– Insertion – extra base
– Deletion – removal of a base
• Frameshift mutations –
nonfunctional proteins

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