From Gene

to Protein

How Genes
Work

AP Biology 2007-2008
 What do genes code for?
 How does DNA code for cells & bodies?
 how are cells and bodies made from the
instructions in DNA

DNA proteins cells bodies

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 The “Central Dogma”
 Flow of genetic information in a cell
 How do we move information from DNA to proteins?

p ti on ti on
c ri l a
trans trans
DNA RNA protein trait

replication

AP Biology
 1941 | 1958
Beadle & Tatum
one gene : one enzyme hypothesis

George Beadle

Edward Tatum
"for their discovery that genes act by
AP Biology regulating definite chemical events"
 a
From gene to protein a a
a
nucleus cytoplasm a
a
a
a
a
a
a
transcription translation a a
DNA mRNA protein
a
a
a

a
a a
a a
a
ribosome

trait
AP Biology
 Transcription
from
DNA language
to
RNA language

AP Biology 2007-2008
 RNA
 ribose sugar
 N-bases
uracil instead of thymine
U : A

C : G

 single stranded
 lots of RNAs
 mRNA, tRNA, rRNA

transcription
DNA
AP Biology RNA
 Transcription
 Making mRNA
 transcribed DNA strand = template strand
 enzyme
 RNA polymerase

coding strand

A G C A T C G T A
3
5 G
A
T
G
C
A
T C
T
A
T
C A T C A G
T
DNA G T A 3 A
C T
G C A U C G U T
G 5
C unwinding
3 G T A G C A
rewinding

mRNA 5 RNA polymerase template strand

AP Biology
build RNA 53
 Initiation
 Promoter region
 binding site before beginning of gene
 TATA box binding site

 binding site for RNA polymerase

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 Elongation
A
 Match RNA bases to DNA C U
G
bases on one of the DNA G A

strands U G
C
U U
C G
A
A C
U
A
AG
C
U
5' RNA
A 3'
A C C polymerase G

T G G T A C A G C T A G T C A T CG T A C CG T

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 Termination
 Eventually the RNA transcript is
released and the polymerase detaches
(complete mechanism still not fully
known)

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 Eukaryotic genes have junk!
 Eukaryotic genes are not continuous
 exons = the real gene introns
 expressed / coding DNA come out!
 introns = the junk
 inbetween sequence

intron = noncoding (inbetween) sequence

eukaryotic DNA
exon = coding (expressed) sequence

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 mRNA splicing
 Post-transcriptional processing
 eukaryotic mRNA needs work after transcription
 primary transcript = pre-mRNA
 mRNA splicing
 edit out introns

 make mature mRNA transcript

intron = noncoding (inbetween) sequence
~10,000 base
eukaryotic DNA
exon = coding (expressed) sequence
pre-mRNA
primary mRNA
transcript
~1,000 base
AP Biology mature mRNA spliced mRNA
transcript
 Splicing must be accurate
 No room for mistakes!
 a single base added or lost throws off the
reading frame

AUGCGGCTATGGGUCCGAUAAGGGCCAU
AUGCGGUCCGAUAAGGGCCAU
AUG|CGG|UCC|GAU|AAG|GGC|CAU
Met|Arg|Ser|Asp|Lys|Gly|His
AUGCGGCTATGGGUCCGAUAAGGGCCAU
AUGCGGGUCCGAUAAGGGCCAU
AUG|CGG|GUC|CGA|UAA|GGG|CCA|U
AP Biology Met|Arg|Val|Arg|STOP|
 RNA splicing enzymes
snRNPs
snRNA
exon intron exon
5' 3'

spliceosome
5' 3'

lariat

5' 3'

exon exon
mature mRNA excised
5' 3' intron
AP Biology
 Alternative splicing
 Alternative mRNAs produced from same gene
 when is an intron not an intron…
 different segments treated as exons

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 More post-transcriptional processing
 Need to protect mRNA on its trip from
nucleus to cytoplasm
 enzymes in cytoplasm attack mRNA
 protect the ends of the molecule
 add 5 GTP cap
 add poly-A tail
 longer tail, mRNA lasts longer: produces more protein

A tai l
-
' p oly 3'
3 A
A
A
A A
mRNA A’s
cap 250
50-
5' P P
5' G P

AP Biology
 a
From gene to protein a a
a
nucleus cytoplasm a
a
a
a
a
a
a
transcription translation a a
DNA mRNA protein
a
a
a

a
a a
a a
a
ribosome

trait
AP Biology
 Translation
from
nucleic acid language
to
amino acid language

AP Biology 2007-2008
 How does mRNA code for proteins?
DNA TACGCACATTTACGTACGCGG
4 ATCG

mRNAAUGCGUGUAAAUGCAUGCGCC
4 AUCG
?
protein Met Arg Val Asn Ala Cys Ala
20
How can you code for 20 amino acids
AP Biology with only 4 nucleotide bases (A,U,G,C)?
 mRNA codes for proteins in triplets

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 1960 | 1968
Cracking the code Nirenberg & Khorana

 Crick
 determined 3-letter (triplet) codon system
WHYDIDTHEREDBATEATTHEFATRAT
 Nirenberg (47) & Khorana (17)
 determined mRNA–amino acid match
 added fabricated mRNA to test tube of

ribosomes, tRNA & amino acids

 created artificial UUUUU… mRNA
 found that UUU coded for phenylalanine
AP Biology
 The code
 Code for ALL life!
 strongest support
for a common origin
for all life
 Code is redundant
 several codons for
each amino acid
 3rd base “wobble”

 Start codon
 AUG
 methionine
 Stop codons
 UGA, UAA, UAG
AP Biology
 How are the codons matched to
amino acids?
3 5
DNA TACGCACATTTACGTACGCGG

5 3
AUGCGUGUAAAUGCAUGCGCC
mRNA
3 5 codon
UAC
tRNA GCA
Met
CAU anti-codon
amino Arg
acid Val
AP Biology
 a
From gene to protein a a
a
nucleus cytoplasm a
a
a
a
a
a
a
transcription translation a a
DNA mRNA protein
a
a
a

a
a a
a a
a
ribosome
aa

trait
AP Biology
 Transfer RNA structure
 “Clover leaf” structure
 anticodon on “clover leaf” end
 amino acid attached on 3 end

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 Loading tRNA
 Aminoacyl tRNA synthetase
 enzyme which bonds amino acid to tRNA
 bond requires energy
 ATP  AMP
 bond is unstable
 so it can release amino acid at ribosome easily

Trp C=O Trp C=O Trp

OH H2O
OH O

C
=O
O
activating
enzyme

tRNATrp AC C
anticodon
UGG mRNA
tryptophan attached
AP Biology to tRNATrp tRNATrp binds to UGG
condon of mRNA
 Ribosomes
 Facilitate coupling of
tRNA anticodon to
mRNA codon
 organelle or enzyme?
 Structure
 ribosomal RNA (rRNA) & proteins
 2 subunits

 large
 small E P A

AP Biology
 Ribosomes
 A site (aminoacyl-tRNA site)
 holds tRNA carrying next amino acid to
be added to chain
 P site (peptidyl-tRNA site)
 holds tRNA carrying growing
polypeptide chain Met

 E site (exit site)

 empty tRNA
leaves ribosome 5' U A C
A U G
from exit site 3'
E P A
AP Biology
 Building a polypeptide
 Initiation
 brings together mRNA, ribosome
subunits, initiator tRNA
 Elongation
 adding amino acids based on
codon sequence
 Termination
 end codon 3 2 1
Leu Val release
Ser factor
Met Met
Met Met Leu Leu Leu Ala
Trp
tRNA
C
A

U AC UAC GAC A U A C GA C U AC G A C
5' CUGAA U 5'
A U G CU G AU 5' A U G C UG AAU 5'
AU G C U G
AA U 3'
mRNA A U G 3' 3' 3' A CC
U GG U A A
E P A 3'
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 Destinations:
Protein targeting 
secretion

nucleus
 Signal peptide  mitochondria

chloroplasts
 address label 
cell membrane

cytoplasm
start of a secretory pathway

etc…

AP Biology
 RNA polymerase
DNA

Can you tell amino

acids
the story? exon intron
tRNA
pre-mRNA 5' GTP cap

mature mRNA
aminoacyl tRNA
poly-A tail synthetase

large ribosomal subunit 3'

polypeptide

5'
tRNA
small ribosomal subunit
E P A

AP Biology ribosome
 The Transcriptional unit (gene?)
enhancer
1000+b exons
20-30b

3' transcriptional unit (gene) 5'

RNA
TATA
polymerase DNA

DNA
introns
promoter

5' 3'
pre-mRNA

5' 3'
AP Biology GTP mature mRNA AAAAAAAA