14

From DNA to Protein: Gene

Expression
14 From DNA to Protein: Gene Expression

•14.2 How Does Information Flow from Genes to Proteins?

•14.3 How Is the Information Content in DNA Transcribed to Produce
RNA?
•14.4 How Is Eukaryotic DNA Transcribed and RNA Processed?
•14.5 How Is RNA Translated into Proteins?
•14.6 What Happens to Polypeptides after Translation?
14.2 How Does Information Flow from Genes to Proteins?

→ RNA → protein
Gene expression to form a specific polypeptide occurs in two steps:

DNA
1. Transcription— 2. Translation—
copies information from a converts RNA sequence
DNA sequence (a gene) to amino acid sequence
to a complementary RNA of a polypeptide
sequence
14.2 How Does Information Flow from Genes to Proteins?
RNA (ribonucleic acid) differs
from DNA:
• Usually one polynucleotide
strand
• The sugar is ribose
• Contains uracil (U) instead of
thymine (T)
14.2 How Does Information Flow from Genes to Proteins?

DNA → RNA → protein

1. Transcription 2. Translation

1. Transcription
 DNA-directed synthesis of RNA
 Only template strand of DNA is used
 U (uracil) in DNA is replaced by T
(thymine) in RNA
 mRNA (transcript) is used to direct
synthesis of polypeptides
 mRNA travels from nucleus to
cytoplasm carrying information as
codons.
 mRNA forms as a complementary
copy of one DNA strand in a gene.
14.2 How Does Information Flow from Genes to Proteins?

DNA → RNA → protein

1. Transcription 2. Translation

2. Translation
 Synthesis of polypeptides
 Takes place at ribosome
Three kinds of RNA in protein synthesis
• Messenger RNA (mRNA)—carries
copy of a DNA sequence to site of
protein synthesis at the ribosome
• Transfer RNA (tRNA)—carries amino
acids for polypeptide assembly
• Ribosomal RNA (rRNA)—RNA
component of ribosome;catalyzes
peptide bonds
14.2 How Does Information Flow from Genes to Proteins?

tRNA: an adapter molecule that can bind amino acids, and recognize a
nucleotide sequence.
tRNA: contains anticodons complementary to the codons in mRNA.
tRNA molecules carrying amino acids line up on mRNA in proper
sequence for the polypeptide chain—translation.
14.3 How Is the Information Content in DNA Transcribed to
Produce RNA?

Transcription occurs in three phases:

• Initiation
• Elongation
• Termination
Figure 14.4 DNA Is Transcribed to Form RNA (A)

Initiation requires a promoter—a special

sequence of DNA.
RNA polymerase binds to the promoter.
Promoter tells RNA polymerase where to
start and which strand of DNA to transcribe.
Part of each promoter is the initiation site.
Figure 14.4 DNA Is Transcribed to Form RNA (B)

Elongation: RNA
polymerase unwinds
DNA about ten base
pairs at a time; reads
template in 3′ to 5′
direction.
The RNA transcript is
antiparallel to the DNA
template strand, and
adds nucleotides to its 3′
end.
Errors can occur 1/100000 –
RNA is short lived

Termination: Is specified by
a specific DNA base
sequence.
Mechanisms of termination
are complex and varied.
14.3 How Is the Information Content in DNA Transcribed to Produce RNA?

The genetic code: Specifies which amino acids

will be used to build a protein
Codon: A sequence of three bases—each codon
specifies a particular amino acid.
Start codon: AUG—initiation signal for translation.
Stop codons: UAA, UAG, UGA—stop translation
and polypeptide is released.
The genetic code is nearly universal: The codons
that specify amino acids are the same in all
organisms.
14.4 How Is Eukaryotic DNA Transcribed and the RNA Processed?

Prokaryotes and eukaryotes differ in gene

structure—in the organization of
nucleotide sequences.

In eukaryotes a
nucleus separates
transcription and
translation
14.4 How Is Eukaryotic DNA Transcribed and the RNA Processed?

Each eukaryote gene has one promoter

to which RNA polymerase binds. At the
other end of the gene there is a
terminator sequence to signal end of
transcription.
14.4 How Is Eukaryotic DNA Transcribed and the RNA Processed?

Eukaryotic genes may have noncoding sequences

—introns (intervening regions).
The coding sequences are exons (expressed
regions).
Introns and exons appear in the primary mRNA
transcript—pre-mRNA; introns are removed from
the final mRNA.
14.4 How Is Eukaryotic DNA Transcribed and the RNA Processed?

In the nucleus, pre-mRNA is modified at both ends –

two processes:
1. G cap is added at the 5′ end (modified guanosine
triphosphate)—facilitates mRNA binding to
ribosome.
G cap protects mRNA from being digested by
ribonucleases.
14.4 How Is Eukaryotic DNA Transcribed and the RNA Processed?

2. Poly A tail added at 3′ end.

AAUAAA sequence after last codon is a signal for
an enzyme to cut the pre-mRNA; then another
enzyme adds 100 to 300 adenines—the “tail.”
May assist in export from nucleus; important for
stability of mRNA.
14.5 How Is RNA Translated into Proteins?

After transcription, mature mRNA carries

the codons required to build proteins

mRNA leaves the nucleus

to begin translation

Translation requires
transfer RNA (tRNA) to
read mRNA codons
correctly
14.5 How Is RNA Translated into Proteins?

tRNA (transfer RNA) translates the

information from mRNA codons into
protein
Two key elements must happen:
1. tRNAs must read mRNA codons
correctly
2. tRNAs must deliver amino acids
corresponding to each codon
14.5 How Is RNA Translated into Proteins?

Three functions of tRNA:

1.It binds to an amino acid, and is then
“charged”
2.It associates with mRNA molecules
3.It interacts with ribosomes
14.5 How Is RNA Translated into Proteins?

Charging tRNA with the correct amino

acids requires activating enzyme:
aminoacyl-tRNA synthetase
Each enzyme is highly specific for one
amino acid and its corresponding tRNA

The enzymes have three-part active

sites: They bind a specific amino acid,
a specific tRNA, and ATP.
14.5 How Is RNA Translated into Proteins?

Ribosome: holds Ribosomes have two

mRNA and subunits, large and
charged tRNAs in small. Ribosomal
the correct subunits are held
positions to allow together by ionic and
assembly of hydrophobic forces
polypeptide chain. (not covalent bonds).
Ribosomes are not
When not active in
specific, they can
translation, the
make any type of
subunits exist
protein.
separately.

small

big
14.5 How Is RNA Translated into Proteins?

Large subunit has three tRNA binding sites:

• A (amino acid) site binds

with anticodon of charged
tRNA
• P (polypeptide) site is where
tRNA adds its amino acid to
the growing chain
• E (exit) site is where tRNA
sits before being released
from the ribosome.
14.5 How Is RNA Translated into Proteins?

Like transcription, translation also occurs

in three steps:
• Initiation
• Elongation
• Termination
14.5 How Is RNA Translated into Proteins?

mRNA start codon is AUG

Initiation:
Large subunit joins the
An initiation complex
complex; the charged tRNA
forms—a charged is now in the P site of the
tRNA and small large subunit
ribosomal subunit,
both bound to
mRNA.

In eukaryotes the
small subunit binds
to the 5′ cap on the
mRNA and moves
until it reaches the
start codon.
14.5 How Is RNA Translated into Proteins?

Elongation: The second

charged tRNA enters the
A site.
Large subunit catalyzes two
reactions:
• It breaks bond between
tRNA in P site and its
amino acid
• Peptide bond forms
between that amino acid
and the amino acid on
tRNA in the A site
Figure 14.16 The Elongation of Translation (Part 2)
Figure 14.17 The Termination of Translation (Part 1)

Termination: Translation
ends when a stop
codon enters the A site.
Stop codon binds a
protein release factor—
allows hydrolysis of
bond between
polypeptide chain and
tRNA on the P site.
Polypeptide chain
separates from the
ribosome
Figure 14.17 The Termination of Translation (Part 2)
14.6 What Happens to Polypeptides after Translation?

Posttranslational aspects of protein synthesis:

Polypeptide emerges from the ribosome and
folds into its 3-D shape.
Amino acid sequence gives a set of
instructions to the polypeptide:
“Finish translation and send to an organelle,
or remain in the cytosol”

.
14.6 What Happens to Polypeptides after Translation?

Protein modification after translation: Post-translational modification:

• Proteolysis: cutting of polypeptide chain

• Glycosylation: addition of sugars to form glycoproteins
• Phosphorylation: addition of phosphate groups
14.6 What Happens to Polypeptides after Translation?

Protein modification after translation