Dr.

Ali Asgher Shuja

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 PROTEIN
SYNTHESIS

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 Protein Synthesis
 The production (synthesis) of
polypeptide chains (proteins)
 Two phases:
Transcription & Translation
 mRNA must be processed before it
leaves the nucleus of eukaryotic cells

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DNA  RNA  Protein
DNA

Transcription

mRNA

Ribosome

Translation

Protein

Prokaryotic Cell 4
 DNA  RNA  Protein
Nuclear
DNA membrane

Transcription
Pre-mRNA
Eukaryotic RNA Processing
Cell mRNA

Ribosome

Translation

Protein
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Pathway to Making a
Protein
DNA

mRNA

tRNA (ribosomes)

Protein
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Nucleic Acids

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 DNA or Protein?
 Walter Sutton discovered
chromosomes were made of
DNA and Protein
 However, scientists were
NOT sure which one (protein
or DNA) was the actual
genetic material of the cell
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 DNA!
 Frederick Griffith
in 1928 showed the
DNA was the cell’s
genetic material
 Watson & Crick in
the 1950’s built the
1st model of DNA
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 Structure of DNA
 DNA is made of subunits called
nucleotides
 DNA nucleotides are composed of
a phosphate, deoxyribose sugar,
and a nitrogen-containing base
 The 4 bases in DNA are: adenine
(A), thymine (T), guanine (G),
and cytosine (C)
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DNA Nucleotide
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 Base Pairing Rule
• Watson and Crick showed that DNA
is a double helix
• A (adenine) pairs with T (thymine)
• C (cytosine) pairs with G (guanine)

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 Nitrogen Rings
• Purines have single rings of
carbon-nitrogen (G, A)
• Pyrimidines have double carbon-
nitrogen rings (C, T)
• This is called complementary
base pairing because a purine is
always paired with a pyrimidine

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 5’ to 3’ Sugars
.
When the DNA double helix
unwinds, it resembles a ladder
The sides of the ladder are the
sugar-phosphate backbones
The rungs of the ladder are
the complementary paired bases
The two DNA strands are anti-
parallel (they run in opposite
directions)

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 Anti-
Parallel
Strands
of DNA

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 DNA
Replication

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Steps in DNA Replication
Occurs when chromosomes duplicate
(make copies)
An exact copy of the DNA is produced
with the aid of the enzyme DNA
polymerase
Hydrogen bonds between bases break
and enzymes “unzip” the molecule
Each old strand of nucleotides serves
as a template for each new strand
New nucleotides move into
complementary positions are joined by
DNA polymerase
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 Two New,
Identical
DNA
Strands
Result
from
Replication

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Another View of Replication

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RNA

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RNA Differs from DNA
1. RNA has a sugar ribose
DNA has a sugar deoxyribose
2. RNA contains the base uracil
(U)
DNA has thymine (T)
3. RNA molecule is single-
stranded
DNA is double-stranded

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Structure of RNA

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.
Three Types of RNA
• Messenger RNA (mRNA) carries
genetic information to the
ribosomes
• Ribosomal RNA (rRNA), along
with protein, makes up the
ribosomes
• Transfer RNA (tRNA) transfers
amino acids to the ribosomes
where proteins are synthesized
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Making a
Protein

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 Genes & Proteins
 Proteins are made of amino
acids linked together by peptide
bonds
 20 different amino acids exist
 Amino acids chains are called
polypeptides
 Segment of DNA that codes for
the amino acid sequence in a
protein are called genes 25
 Two Parts of Protein
Synthesis
 Transcription makes an RNA
molecule complementary to a
portion of DNA
 Translation occurs when the
sequence of bases of mRNA
DIRECTS the sequence of amino
acids in a polypeptide
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 Genetic Code
 DNA contains a triplet code
 Every three bases on DNA stands
for ONE amino acid
 Each three-letter unit on mRNA is
called a codon
 Most amino acids have more than
one codon!
 There are 20 amino acids with a
possible 64 different triplets
 The code is nearly universal among
living organisms
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Transcription

Translation

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Overview of Transcription
 During transcription in the
nucleus, a segment of DNA
unwinds and unzips, and the DNA
serves as a template for mRNA
formation
 RNA polymerase joins the RNA
nucleotides so that the codons in
mRNA are complementary to the
triplet code in DNA
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 Steps in Transcription
 The transfer of information in the nucleus
from a DNA molecule to an RNA molecule
 Only 1 DNA strand serves as the
template
 Starts at promoter DNA (TATA box)
 Ends at terminator DNA (stop)
 When complete, pre-RNA molecule is
released

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Transcription

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What is the enzyme
responsible for the
production of the
mRNA molecule?

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 RNA Polymerase
 Enzyme found in the nucleus
 Separates the two DNA
strands by breaking the
hydrogen bonds between the
bases
 Then moves along one of the
DNA strands and links RNA
nucleotides together
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DNA

RNA Polymerase

pre-mRNA

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 Question:
 What would be the
complementary RNA strand
for the following DNA
sequence?

DNA 5’-GCGTATG-3’
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 Answer:

• DNA 5’-GCGTATG-3’
• RNA 3’-CGCAUAC-5’

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 Processing Pre-mRNA
• Also occurs in the nucleus
• Pre-mRNA made up of segments
called introns & exons
• Exons code for proteins, while
introns do NOT!
• Introns spliced out by splicesome-
enzyme and exons re-join
• End product is a mature RNA
molecule that leaves the nucleus to
the cytoplasm
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 RNA Processing
pre-RNA molecule
exon intro exon intron exon
n

intron intron

exon exon exon

splicesome splicesome

exon exon exon

Mature RNA molecule 40

 Messenger RNA (mRNA)
• Carries the information for a
specific protein
• Made up of 500 to 1000
nucleotides long
• Sequence of 3 bases called codon
• AUG – methionine or start codon
• UAA, UAG, or UGA – stop codons

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 Messenger RNA (mRNA)
start
codon

mRNA A U G G G C U C C A U C G G C G C A U A A

codon 1 codon 2 codon 3 codon 4 codon 5 codon 6 codon 7

protein methionine glycine serine isoleucine glycine alanine stop

codon

Primary structure of a protein

aa1 aa2 aa3 aa4 aa5 aa6

peptide bonds 42
 Transfer RNA (tRNA)
• Made up of 75 to 80 nucleotides long
• Picks up the appropriate amino acid
floating in the cytoplasm
• Transports amino acids to the mRNA
• Have anticodons that are
complementary to mRNA codons
• Recognizes the appropriate codons on
the mRNA and bonds to them with
H-bonds
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Transfer RNA (tRNA)
amino acid
attachment site methionine amino acid

U A C
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anticodon
 Ribosomal RNA (rRNA)
• Made up of rRNA
is 100 to 3000
nucleotides long
• Made inside the
nucleus of a cell
• Associates with
proteins to form
ribosomes
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 Ribosomes
• Made of a large and small
subunit
• Composed of rRNA (40%) and
proteins (60%)
• Have two sites for tRNA
attachment --- P and A

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 Ribosomes

Large
subunit
P A
Site Site

mRNA
A U G C U A C U U C G
Small
subunit 47
 Translation
• Synthesis of proteins in the
cytoplasm

• Involves the following:

1. mRNA (codons)
2. tRNA (anticodons)
3. ribosomes
4. amino acids
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 Translation
• Three steps:
1. initiation: start codon (AUG)
2. elongation: amino acids linked
3. termination: stop codon
(UAG, UAA, or UGA).

Let’s Make a Protein !

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 mRNA Codons Join the
Ribosome

Large
subunit
P A
Site Site

mRNA
A U G C U A C U U C G
Small subunit 50
 Initiation
aa2
aa1

2-tRNA
1-tRNA
G A U
anticodon U A C
hydrogen A U G C U A C U U C G A
bonds codon mRNA
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 Elongation
peptide bond
aa3
aa1 aa2

3-tRNA

1-tRNA 2-tRNA G A A
anticodon U A C G A U
hydrogen A U G C U A C U U C G A
bonds codon mRNA
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 aa1 peptide bond
aa3
aa2

1-tRNA

U A C 3-tRNA
(leaves)
2-tRNA G A A

G A U
A U G C U A C U U C G A
mRNA
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Ribosomes move over one codon
 peptide bonds
aa1 aa4

aa2 aa3

4-tRNA

2-tRNA 3-tRNA G C U

G A U G A A
A U G C U A C U U C G A A C U
mRNA
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 peptide bonds
aa1 aa4
aa2

aa3

2-tRNA
4-tRNA
G A U
(leaves) 3-tRNA G C U

G A A
A U G C U A C U U C G A A C U
mRNA
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Ribosomes move over one codon
 peptide bonds aa5
aa1
aa2
aa4
aa3

5-tRNA

U G A
3-tRNA 4-tRNA

G A A G C U
G C U A C U U C G A A C U
mRNA
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 aa1 peptide bonds aa5
aa2
aa3
aa4

5-tRNA

3-tRNA U G A
G A A 4-tRNA

G C U
G C U A C U U C G A A C U
mRNA
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Ribosomes move over one codon
 aa5
aa4 aa199
Termination
aa3 primary aa200
structure
aa2 of a protein

aa1
terminator
200-tRNA
or stop
codon
A C U C A U G U U U A G
mRNA
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 End Product –The Protein!
• The end products of protein
synthesis is a primary structure of
a protein
• A sequence of amino acid bonded
together by peptide bonds

aa5
aa3 aa4
aa2 aa199

aa1 aa200
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