Gene Expression
How proteins are made.
W
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K
• what monomers make up proteins?
• what monomers make up nucleic acids
(DNA and RNA)?
T
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E
T
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Nucleic Acids
Proteins
Primarily DNA, RNA
Thousands of different
proteins
Made up of four
different bases
Made up of more than
than 20 amino acids
So, how does relatively simple-sounding DNA
contain the information for building thousands of
different proteins?
Codons
A “codon” is a sequence of three bases in DNA
and RNA. Each codon codes for a different
amino acid.
This mRNA strand:
codes for
these amino
acids:
met cys
glu
leu
trp
The Genetic Code
All 20 amino acids are coded for. Redundancy of codes is
one protection against mutations.
The Gene Concept
• A “gene” is a segment of DNA that
codes for a specific protein.
• Only one side of the DNA double-helix
(the “sense” or “coding” strand) contains
the actual gene.
• Genes are defined by promotor and
terminator sequences in the DNA.
Eukaryotic gene structure
exons
DNA
promoter
introns
A typical eukaryotic gene consists of sequences of DNA called
exons, which code for the amino acids of a protein (medium
blue),
and intervening sequences called introns (dark blue), which do
not. The promoter (light blue) determines where RNA
polymerase
will begin transcription.
A small protein is 30 amino acids long. How
many nucleotides are needed to code for it?
25%
1.
2.
3.
4.
25%
25%
2
3
25%
30
60
90
Depends on which
amino acids.
1
4
The same protein that is 30 amino acids long
needs how many codons to code for it?
68%
1.
2.
3.
4.
30
60
90
Depends on the
amino acids it is made
of.
19%
11%
2%
1
2
3
4
Transcription
• DNA stays in the nucleus.
• To get information out of one gene on a
strand of DNA, the gene must be
transcribed.
• An mRNA copy of a gene leaves the
nucleus, so the original information
(DNA) remains intact in the nucleus.
RNA
• RNA is a single-stranded nucleic acid.
• RNA contains the bases adenine, uracil,
guanine, and cytosine.
• RNA contains the sugar ribose in its
sugar-phosphate backbone.
• Which of these is TRUE about RNA:
• RNA has uracil instead of thymine.
• RNA is a protein.
• RNA is a single strand instead of a
double-helix.
• RNA never leaves the nucleus.
W
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T
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E
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gene
DNA
(nucleus)
(a) Transcription
messenger
RNA
ribosome
protein
(cytoplasm)
Transcription of the
gene produces an
mRNA with a
nucleotide sequence
complementary to one
of the DNA strands.
Notice that
transcription takes
place in the
nucleus.
(a) Initiation
DNA
gene 1
gene 2
gene 3
RNA
polymerase
DNA
promoter
RNA polymerase binds to the promoter region of DNA near the
beginning of a gene, separating the double helix near the
promoter.
(b) Elongation
RNA
DNA template strand
RNA polymerase travels along the DNA template strand (blue),
catalyzing the addition of ribose nucleotides into an RNA
molecule (pink). The nucleotides in the RNA are complementary
to the template strand of the DNA.
(c) Termination
termination signal
At the end of a gene, RNA polymerase encounters a DNA
sequence called a termination signal. RNA polymerase detaches
from the DNA and releases the mRNA molecule.
(d) Conclusion of transcription
mRNA
After termination, the DNA completely rewinds into a double helix.
The RNA molecule is free to move from the nucleus to the
cytoplasm for translation, and RNA polymerase may move to
another gene and begin transcription once again.
RNA synthesis and processing in eukaryotes
DNA
transcription
initial
RNA transcript
add RNA cap and tail
cap
tail
RNA splicing
completed
mRNA
introns
cut out
and
broken
down
to cytoplasm for translation
RNA polymerase transcribes both the exons and introns, producing a long
RNA molecule. Enzymes in the nucleus then add further nucleotides at the
beginning (cap) and end (tail) of the RNA transcript. Other enzymes cut out
the RNA introns and splice together the exons to form the true mRNA, which
moves out of the nucleus and is translated on the ribosomes.
gene
RNA
molecules
DNA
Transcription begins when:
1. RNA polymerase
finds a start
codon
2. RNA polymerase
finds a promoter
sequence
3. RNA polymerase
finds a ribosome
33%
1
33%
2
33%
3
Base-pair matching, DNA
mRNA
DNA (ns)
DNA (sense)
mRNA
A
T
A
G
C
G
T
A
U
A
T
A
C
G
C
G
C
G
T
A
U
• The enzyme that assembles RNA bases
to make mRNA is: _________________
RNA Polymerase
• This enzyme begins reading DNA at the
promoter
____________
sequence of a gene and
ends at the ___________
sequence.
terminator
• True or False: The entire DNA strand
must be “unzipped” for transcription to
take place.
W
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T
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G
E
T
H
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Translation
• Once the gene has been transcribed into
mRNA, the message must be translated
to build a protein.
• Ribosomes (made of rRNA) “read” the
mRNA message and use the information
to assemble amino acids.
gene
DNA
(nucleus)
(cytoplasm)
Notice that
translation takes
place outside the
nucleus, at the
ribosomes.
messenger
RNA
(b) Translation
ribosome
protein
Translation of the mRNA
produces a protein
molecule with an amino
acid sequence determined
by the nucleotide
sequence in the mRNA.
The players:
• mRNA: Carries the encoded instructions
for building a protein.
• Ribosome (rRNA & protein structures):
these act like enzymes to catalyze
protein assembly.
• tRNA: Transport RNA molecules that
carry amino acids from the cytoplasm to
the ribosome.
To what class of molecules does tRNA
belong?
100%
1.
2.
3.
4.
5.
Proteins
Carbohydrates
Lipids
Nucleic acids
Depends on which
amino acid it
carries.
0%
1
0%
2
0%
3
0%
4
5
W
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K
• What other molecule have we
encountered has active sites and acts as
a catalyst? How is a ribosome like this
molecule? How is it different?
T
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Initiation:
amino acid
met
initiation
complex
methionine
tRNA
small
ribosomal
subunit
A tRNA with an attached methionine amino acid binds to a
small ribosomal subunit, forming an initiation complex.
Initiation:
met
tRNA
mRNA
The initiation complex binds to an mRNA molecule.
The methionine (met) tRNA anticodon (UAC) base-pairs
with the start codon (AUG) of the mRNA.
Initiation:
catalytic site
first tRNA
binding
site
second tRNA binding site
large
ribosomal
subunit
The large ribosomal subunit binds to the small subunit.
The methionine tRNA binds to the first tRNA site on
the large subunit.
Elongation:
catalytic site
The second codon of mRNA (GUU) base-pairs with the
anticodon (CAA) of a second tRNA carrying the amino acid
valine (val). This tRNA binds to the second tRNA site on the
large subunit.
Elongation:
peptide
bond
The catalytic site on the large subunit catalyzes the
formation of a peptide bond linking the amino acids methionine
and valine. The two amino acids are now attached to the tRNA
in the second binding position.
Is this hydrolysis or dehydration synthesis?
Elongation:
initiator
tRNA detaches
catalytic site
ribosome moves one codon to right
The “empty” tRNA is released and the ribosome moves down
the mRNA, one codon to the right. The tRNA that is attached to
the two amino acids is now in the first tRNA binding site and
the second tRNA binding site is empty.
Elongation:
catalytic site
The third codon of mRNA (CAU) base-pairs with the
anticodon (GUA) of a tRNA carrying the amino acid histidine
(his). This tRNA enters the second tRNA binding site on the
large subunit.
Elongation:
The catalytic site forms a new peptide bond between valine
and histidine. A three-amino-acid chain is now attached to
the tRNA in the second binding site. The tRNA in the first site
leaves, and the ribosome moves one codon over on the mRNA.
Termination:
completed
peptide
stop codon
This process repeats until a stop codon is reached; the mRNA
and the completed peptide are released from the ribosome,
and the subunits separate.
The ribosome has just bonded a series of
amino acids into a chain. What has it built?
80%
1.
2.
3.
4.
An amino acid.
A protein.
A nucleic acid.
Impossible to tell at
this point.
20%
0%
1
0%
2
3
4
• When a tRNA leaves the ribosome, it
goes off and finds another amino acid in
the cell. Where do amino acids in human
cells originally come from? Where do
they come from in plant cells?
W
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T
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gene
(a) DNA
complementary
DNA strand
etc.
template DNA
strand
etc.
codons
(b) mRNA
etc.
anticodons
(c) tRNA
etc.
amino acids
(d) protein
methionine
glycine
valine
etc.
direction of transcription
RNA
polymerase
DNA
mRNA
protein
ribosome
DNA to mRNA to Protein
DNA (Sense)
mRNA
T
A
C
G
G
T
A
G
A
A
U
G
C
C
A
U
C
U
Amino Acids
methionine
(start)
proline
serine
DNA to mRNA to Protein
DNA (Sense) mRNA (sense) Amino Acids
C
A
A
T
G
A
A
C
T
G
U
U
A
C
valine
threonine
U
U
G
A
stop codon
Practice Transcription and Translation:
http://learn.genetics.utah.edu/content/begin/dna/transcribe/
Translation begins when:
1. The ribosome finds
a promoter
sequence.
2. The ribosome finds
a start codon.
3. The ribosome
breaks apart.
60%
40%
0%
1
2
3
The role of the ribosome is:
1. Interpret mRNA
and build proteins.
2. Construct mRNA.
3. Replicate DNA.
4. Facilitate cell
division.
80%
20%
0%
1
2
3
0%
4
The role of tRNA is:
1. Transcribe DNA and
move mRNA out of
the nucleus.
2. Bind to the ribosome
and mRNA chain
together.
3. Carry amino acids to
the ribosome.
4. Replace T with U
when transcribing
mRNA.
80%
20%
0%
1
2
0%
3
4
If genes code for proteins, what
codes for enzymes?
1.
2.
3.
4.
Genes
Non-coding DNA
Other proteins
Nothing. They’re
manufactured in
the smooth ER.
60%
40%
0%
1
2
0%
3
4
• Write out the mRNA strand that would be
formed from this DNA segment:
CATATGGGCTTATAC
• If the segment doesn’t include the start or
stop codon, how many amino acids does it
code for?
W
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T
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E
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W
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• Suppose a segment of DNA contains the K
triplet ACG, and a mutation changes it to
ACT. Would that cause a change in the
resulting amino acid chain? Use your
knowledge of transcription and
translation to find the answer.
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Gene Regulation
• All cells in the human body have the
same DNA and the same set of genes,
yet different cells look different and do
different jobs.
• Cells have systems to regulate which
genes are “turned on” (transcribed) and
which are not.
DNA
1 transcription
rRNA
pre-mRNA
tRNA
+ proteins
2 mRNA
processing
ribosomes
If the active
protein is an
enzyme, it
will catalyze
a chemical
reaction in
the cell.
mRNA
tRNA
amino
acids
3 translation
inactive
protein
4 modification
substrate
active
protein
product
5 degradation
amino
acids
Cells can control
the frequency of
transcription.
Different mRNAs
may be produced
from a single gene.
Cells can control the
stability and rate of
translation of
particular mRNAs.
Cells can regulate
a protein’s activity
by degrading it.
Cells can regulate
a protein’s activity
by modifying it.
(a) Structure of the lactose operon
codes for
repressor protein
R
P O
promoter: RNA
polymerase
binds here
operator: repressor
protein binds here
gene 1
gene 2
gene 3
structural genes that code for
enzymes of lactose metabolism
The lactose operon consists of a regulatory gene, a promoter, an
operator, and three structural genes that code for enzymes
Involved in lactose metabolism. The regulatory gene codes for a
protein, called a repressor, which can bind to the operator site
under certain circumstances.
(b) Lactose absent
RNA
polymerase
transcription blocked
R
P
gene 1
gene 2
gene 3
repressor protein
bound to operator,
overlaps promoter
free repressor
proteins
When lactose is not present, repressor proteins bind to the
operator of the lactose operon. When RNA polymerase binds to
the promoter, the repressor protein blocks access to the
structural
genes, which therefore cannot be transcribed.
(c) Lactose present
RNA polymerase binds
to promoter, transcribes
structural genes
R
O gene 1
gene 2 gene 3
lactosemetabolizing
enzymes
synthesized
lactose bound
to repressor proteins
When lactose is present, it binds to the repressor protein. The
lactose-repressor complex cannot bind to the operator, so
RNA polymerase has free access to the promoter. The RNA
polymerase transcribes the three structural genes coding for
the lactose-metabolizing enzymes.
Recap
• Transcription moves coded information
from DNA to the ribosome by creating
an mRNA copy of a gene.
• In translation, a ribosome “reads” the
mRNA code and uses the information to
assemble a chain of amino acids to
make a protein.