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to Protein
How Genes
Work
AP Biology
2007-2008
AP Biology
large
subunit
Ribosomes
Function
small
subunit
protein production
Structure
0.08m
Ribosomes
Rough
ER
Smooth
ER
AP Biology
Types of Ribosomes
Free ribosomes
suspended in cytosol
synthesize proteins that
function in cytosol
Bound ribosomes
AP Biology
attached to endoplasmic
reticulum
synthesize proteins
for export or
for membranes
membrane proteins
TO:
endoplasmic
reticulum
nucleus
protein
on its way!
DNA
RNA
vesicle
TO:
TO:
TO:
vesicle
ribosomes
TO:
Making Proteins
AP Biology
finished
protein
protein
Golgi
apparatus
lack of an enzyme
Tay sachs
PKU (phenylketonuria)
albinism
metabolic pathway
disease
AP Biologyenzyme 1
Am I just the
sum of my proteins?
disease
enzyme 2
disease
enzyme 3
disease
enzyme 4
AP Biology
Wild-type
Neurospora
create mutations
asexual
spores
Minimal
medium
spores
Growth on
complete
medium
positive control
Select one of
the spores
Test on minimal
medium to confirm
presence of mutation
negative control
Grow on
complete medium
de
i
on
i
t
a
ed
i
f
i
nt
t
Minimal media supplemented only with
mu
experimentals
Choline
Pyridoxine
Minimal
Nucleic
Arginine Riboflavin
amino acid
Niacin
control
Inositol acid
Folic
p-Amino
supplements
AP Biology
Thiamine
acid
benzoic acid
chromosome
gene
cluster 1
gene
cluster 2
arg-E
encoded
enzyme
arg-G arg-H
arg-F
enzyme E
glutamate
gene
cluster 3
enzyme F
ornithine
substrate in
AP biochemical
Biology
pathway
enzyme G
citruline
enzyme H
arginine
arginosuccinate
gene that
was damaged
1941 | 1958
George Beadle
Edward Tatum
AP Biology
DNA
n
o
i
t
p
i
r
c
s
n
tra
replication
AP Biology
RNA
tr
on
i
t
a
l
ans
protein
DNA gets
all the glory,
but proteins do
all the work!
trait
RNA
ribose sugar
N-bases
single stranded
lots of RNAs
DNA
AP Biology
transcription
RNA
Transcription
from
DNA nucleic acid language
to
RNA nucleic acid language
AP Biology
2007-2008
Transcription
Making mRNA
enzyme
coding strand
RNA polymerase
5
DNA
C
G
3
AP Biology
build
RNA 53
A
G
T
A T C
T A
rewinding
mRNA 5
G
C
A G C
C G T
3
G C A U C G U
C
G T A G C A
RNA polymerase
T
A
A
C
T
A G
C T
G
A
T
3
5
unwinding
template strand
Bacterial chromosome
Transcription in
Prokaryotes
Transcription
mRNA
Psssst
no nucleus!
Cell
membrane
Cell wall
AP Biology
2007-2008
Transcription in Prokaryotes
Initiation
Role of promoter
Starting point
where to start reading
start of gene
Template strand
which strand to read
Direction on DNA
always read DNA 35!
build RNA 53
APBiology
Transcription in Prokaryotes
Promoter sequences
enzyme
subunit
RNA polymerase
read DNA 35
bacterial DNA
Promoter
TTGACA TATAAT
35 sequence
10 sequence
RNA polymerase
molecules bound to
bacterial DNA
RNA polymerase
AP Biology
Transcription in Prokaryotes
Elongation
Simple proofreading
1 error/105 bases
make many mRNAs
mRNA has short life
not worth editing!
AP Biology
reads DNA 35
Transcription in Prokaryotes
Termination
RNA GC
hairpin turn
AP Biology
Transcription in Eukaryotes
Transcription
Psssst
DNA cant
leave nucleus!
RNA Processing
Translation
Protein
AP Biology
2007-2008
Prokaryotes
DNA in cytoplasm
circular
chromosome
naked DNA
!
no introns
Eukaryotes
DNA in nucleus
linear
chromosomes
DNA wound on
histone proteins
introns vs. exons
introns
come out!
Transcription in Eukaryotes
RNA polymerase 1
RNA polymerase 2!
AP Biology
RNA polymerase 3!
Transcription in Eukaryotes
Initiation complex
transcription factors
trigger the binding of RNA
polymerase to DNA
AP Biology
Post-transcriptional processing
ail
t
A
ly3'
o
p
3'
AA
AAA
5
50-2
s
0 A
ryotic DNA
exon = coding (expressed) sequence
ary mRNA
transcript
mature
mRNA
AP Biology
transcrip
pre-mRNA
~1,000 bases
spliced mRNA
AP Biology
Whoa! I think
we just broke
a biological rule!
Splicing enzymes
snRNPs
Spliceosome
AP Biology
intron
exon
5'
several snRNPs
recognize splice
site sequence
snRNPs
snRNA
mature mRNA
3'
spliceosome
5'
3'
lariat
5'
exon
5'
3'
exon
3'
excised
Ribozyme
1982 | 1989
RNA as ribozyme
Sidney Altman
AP Biology
Yale
Thomas Cech
U of Colorado
Translation
from
nucleic acid language
to
amino acid language
AP Biology
2007-2008
Bacterial chromosome
Translation in
Prokaryotes
Transcription
mRNA
Translation
Psssst
no nucleus!
protein
Cell
membrane
Cell wall
AP Biology
2007-2008
Translation in Prokaryotes
DNA is in
cytoplasm
no mRNA
editing
ribosomes
read mRNA
as it is being
transcribed
AP Biology
RNA processing
AP Biology
Translation in Eukaryotes
AP Biology
2007-2008
DNA
transcription
mRNA
mRNA leaves
nucleus through
nuclear pores
nucleus
AP Biology
aa
translation
protein
aa
aa
aa
aa
ribosome
cytoplasm
proteins synthesized
by ribosomes using
instructions on mRNA
mRNA
4 AUCG
protein
TACGCACATTTACGTACGCGG
AUGCGUGUAAAUGCAUGCGCC
?
Met Arg Val Asn Ala Cys Ala
20
AP Biology
DNA
TACGCACATTTACGTACGCGG
codon
mRNA
AUGCGUGUAAAUGCAUGCGCC
?
protein
AP Biology
1960 | 1968
Nirenberg & Khorana
Crick
WHYDIDTHEREDBATEATTHEFATRAT
AP Biology
The code
Code is redundant
Why is the
wobble good?
Start codon
AP Biology
AUG
methionine
Stop codons
DNA
transcription
mRNA
aa
translation
protein
aa
aa
aa
aa
ribosome
aa
nucleus
AP Biology
cytoplasm
AP Biology
Ribosomes
Facilitate coupling of
tRNA anticodon to
mRNA codon
organelle or enzyme?
Structure
large
small
E P A
AP Biology
Ribosomes
Met
AP Biology
empty tRNA
leaves ribosome
from exit site
U A C
A U G
5'
3'
Building a polypeptide
Initiation
brings together mRNA, ribosome
subunits, initiator tRNA
Elongation
adding amino acids based on
codon sequence
Termination
3 2 1
end codon
Leu
Val
Met
Met
Met
Met Leu
Leu
Ala
Leu
release
factor
Ser
Trp
tRNA
U AC
5'
C U GA A U
mRNA A U G
3'
E P A
AP Biology
5'
U A C G A C AA
U
A U G CU G
5'
3'
U A C GA C
A U G C UG AAU
3'
5'
U A C G A C AA U
AU G C U G
3'
A CC
U GG U A A
3'
Protein targeting
Destinations:
Signal peptide
address label
AP Biology
secretion
nucleus
mitochondria
chloroplasts
cell membrane
cytoplasm
etc
RNA polymerase
DNA
amino
acids
exon
intron
tRNA
pre-mRNA
5' cap
mature mRNA
polyA tail
aminoacyl tRNA
synthetase
3'
polypeptide
5'
small ribosomal subunit
AP Biology
tRNA
E P A
ribosome
Any Questions??
AP Biology
2007-2008