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Transcript
DNA
Structure
&
Replication
1
2
DNA
• DNA is often
called the
blueprint of life.
• In simple terms,
DNA contains the
instructions for
making proteins
within the cell.
3
Why do we study DNA?
We study DNA for
many reasons,
e.g.,
• its central
importance to all
life on Earth,
• medical benefits
such as cures for
diseases,
• better food crops.
4
Chromosomes and
DNA
• Our genes are on
our
chromosomes.
• Chromosomes
are made up of a
chemical called
DNA.
5
The Shape of the Molecule
• DNA is a very long
polymer.
• The basic shape is
like a twisted ladder
or zipper.
• This is called a
double helix.
6
The Double Helix
Molecule
• Discovered by
James Watson &
Francis Crick
• The DNA double
helix has two
strands twisted
together.
7
One Strand of DNA
• The backbone
of the molecule
is alternating
phosphates and
deoxyribose
sugar
• The teeth are
nitrogenous
bases.
phosphate
deoxyribose
bases
8
Nucleotides
O
O -P O
O
O
O -P O
O
One deoxyribose together
with its phosphate and base
make a nucleotide.
O
O -P O
O
Nitrogenous
base
O
Phosphate
C
C
C
O Deoxyribose
9
One Strand of DNA
nucleotide
• One strand of
DNA is a polymer
of nucleotides.
• One strand of
DNA has many
millions of
nucleotides.
10
Four nitrogenous
bases
DNA has four different bases:
C
• Thymine T
• Adenine A
• Guanine G
• Cytosine
11
Two Stranded DNA
• Remember,
DNA has two
strands that fit
together
something like
a zipper.
• The teeth are
the nitrogenous
bases but why
do they stick
together?
12
C
N
N
C
N
N
C
C
C
O
• The bases attract each
other because of
hydrogen bonds.
• Hydrogen bonds are
weak but there are
millions and millions of
them in a single
molecule of DNA.
• The bonds between
cytosine and guanine
are shown here with
dotted lines
N
Hydrogen Bonds
N
C
N
C
C
O
13
C
N
Hydrogen Bonds,
cont.
• When making
hydrogen bonds,
cytosine always
pairs up with
guanine
• Adenine always
pairs up with
thymine
• Adenine is
bonded to
thymine here
N
O
O
C
C
C C
N
C
14
Chargaff’s Rule:
• Adenine and Thymine
always join together
A
T
• Cytosine and Guanine
always join together
C
G
15
DNA by the Numbers
• Each cell has about 2
m of DNA.
• The average human
has 75 trillion cells.
• The average human
has enough DNA to go
from the earth to the
sun more than 400
times.
• DNA has a diameter of
only 0.000000002 m.
The earth is 150 billion m
or 93 million miles from
the sun.
16
DNA
Replication
17
Replication Facts
• DNA has to be copied
before a cell divides
• DNA is copied during the S
or synthesis phase of
interphase
• New cells will need identical
DNA strands
18
Synthesis Phase (S phase)
• S phase during interphase of the
cell cycle
• Nucleus of eukaryotes
S
DNA replication takes
place in the S phase.
phase
G1
interphase
G2
Mitosis
-prophase
-metaphase
-anaphase
-telophase
19
DNA Replication
• Begins at Origins of Replication
• Two strands open forming Replication
Forks (Y-shaped region)
• New strands grow at the forks
5’ Parental DNA Molecule
3’
3’
Replication
Fork
20
5’
DNA Replication
• As the 2 DNA strands open at
the origin, Replication Bubbles
form
• Prokaryotes (bacteria) have a
single bubble
• Eukaryotic chromosomes have
MANY bubbles
Bubbles
Bubbles
21
DNA Replication
• Enzyme Helicase unwinds
and separates the 2 DNA
strands by breaking the
weak hydrogen bonds
22
Question:
• What would be the
complementary DNA
strand for the following
DNA sequence?
DNA 5’-CGTATG-3’
23
Answer:
DNA 5’-CGTATG-3’
DNA 3’-GCATAC-5’
24
25
PROTEIN
SYNTHESIS
26
DNA
and
Genes
DNA
• DNA contains genes,
sequences of nucleotide
bases
• These Genes code for
polypeptides (proteins)
• Proteins are used to build
cells and do much of the
work inside cells
Genes & Proteins
 Proteins are made of
amino acids linked
together by peptide
bonds
 20 different amino acids
exist
29
Amino Acid Structure
30
Polypeptides
• Amino acid
chains are
called
polypeptides
31
DNA Begins the Process
• DNA is found inside the
nucleus
• Proteins, however, are made
in the cytoplasm of cells by
organelles called ribosomes
• Ribosomes may be free in the
cytosol or attached to the
surface of rough ER
32
Starting with DNA
• DNA ‘s code must be copied
and taken to the cytosol
• In the cytoplasm, this code
must be read so amino acids
can be assembled to make
polypeptides (proteins)
• This process is called
PROTEIN SYNTHESIS
33
RNA
Roles of RNA and DNA
• DNA is the MASTER
PLAN
• RNA is the
BLUEPRINT of the
Master Plan
35
RNA Differs from DNA
• RNA has a sugar ribose
DNA has a sugar deoxyribose
36
Other Differences
•
•
RNA contains the
base uracil (U)
DNA has thymine
(T)
RNA molecule is
single-stranded
DNA is doublestranded
DNA
37
Structure of RNA
38
.
Three Types of RNA
• Messenger RNA (mRNA) copies
DNA’s code & carries the
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
39
Messenger RNA
• Long Straight chain
of Nucleotides
• Made in the Nucleus
• Copies DNA & leaves
through nuclear
pores
• Contains the
Nitrogen Bases A, G,
C, U ( no T )
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
41
codons
Ribosomal RNA (rRNA)
• rRNA is a single
strand 100 to 3000
nucleotides long
• Globular in shape
• Made inside the
nucleus of a cell
• Associates with
proteins to form
ribosomes
• Site of protein
Synthesis
42
The Genetic Code
• A codon designates an amino
acid
• An amino acid may have more
than one codon
• There are 20 amino acids,
but 64 possible codons
• Some codons tell the
ribosome to stop translating
43
The Genetic Code
•Use the
code by
reading from
the center to
the outside
•Example:
AUG codes
for
Methionine
44
Name the Amino Acids
•
•
•
•
•
GGG?
UCA?
CAU?
GCA?
AAA?
45
Remember the
Complementary Bases
On DNA:
A-T
C-G
On RNA:
A-U
C-G
46
Transfer RNA (tRNA)
• Clover-leaf shape
• Single stranded molecule with
attachment site at one end
for an amino acid
• Opposite end has three
nucleotide bases called the
anticodon
47
Transfer RNA
amino acid
attachment site
U A C
anticodon
48
Codons and Anticodons
• The 3 bases of an
anticodon are
complementary to
the 3 bases of a
codon
• Example: Codon ACU
Anticodon UGA
UGA
ACU
49
Transcription
and
Translation
Pathway to Making a
Protein
DNA
mRNA
tRNA (ribosomes)
Protein
51
Protein Synthesis
 The production or
synthesis of polypeptide
chains (proteins)
 Two phases:
Transcription & Translation
 mRNA must be processed
before it leaves the nucleus
of eukaryotic cells
52
DNA  RNA  Protein
Nuclear
membrane
DNA
Transcription
Eukaryotic
Cell
Pre-mRNA
RNA Processing
mRNA
Ribosome
Translation
Protein
53
54
Transcription
• The process of copying
the sequence of one
strand of DNA, the
template strand
• mRNA copies the template
strand
• Requires the enzyme RNA
Polymerase
55
Template Strand
56
Question:
 What would be the
complementary RNA strand
for the following DNA
sequence?
DNA 5’-GCGTATG-3’
57
Answer:
• DNA 5’-GCGTATG-3’
• RNA 3’-CGCAUAC-5’
58
Transcription
• During transcription, RNA
polymerase binds to DNA and
separates the DNA strands
• RNA Polymerase then uses
one strand of DNA as a
template to assemble
nucleotides into RNA
59
Transcription
• Promoters are regions on
DNA that show where RNA
Polymerase must bind to
begin the Transcription of
RNA
• Called the TATA box
• Specific base sequences act
as signals to stop
• Called the termination signal
60
RNA Polymerase
61
mRNA Processing
• After the DNA is
transcribed into RNA,
editing must be done to
the nucleotide chain to
make the RNA functional
• Introns, non-functional
segments of DNA are
snipped out of the chain
62
mRNA Editing
• Exons, segments of DNA that
code for proteins, are then
rejoined by the enzyme ligase
• A guanine triphosphate cap is
added to the 5’ end of the
newly copied mRNA
• A poly A tail is added to the 3’
end of the RNA
• The newly processed mRNA can
then leave the nucleus
63
Result of Transcription
CAP
New Transcript
Tail
64
mRNA Transcript
•mRNA leaves the nucleus
through its pores and goes to
the ribosomes
65
66
67
Translation
• Translation is the process
of decoding the mRNA
into a polypeptide chain
• Ribosomes read mRNA
three bases or 1 codon at
a time and construct the
proteins
68
Transcription
Translation
69
Ribosomes
• Made of a large and small
subunit
• Composed of rRNA (40%)
and proteins (60%)
• Have two sites for tRNA
attachment --- P and A
70
Step 1- Initiation
• mRNA transcript
start codon AUG
attaches to the
small ribosomal
subunit
• Small subunit
attaches to large
ribosomal subunit
mRNA transcript
71
Ribosomes
Large
subunit
P
Site
A
Site
mRNA
Small
subunit
A U G
C U A C U U C G
72
Step 2 - Elongation
• As ribosome moves, two tRNA with
their amino acids move into site A and
P of the ribosome
• Peptide bonds join the amino acids
Initiation
aa1
aa2
2-tRNA
1-tRNA
anticodon
hydrogen
bonds
U A C
A U G
codon
G A U
C U A C U U C G A
mRNA
74
Elongation
peptide bond
aa1
aa3
aa2
3-tRNA
1-tRNA
anticodon
hydrogen
bonds
U A C
A U G
codon
2-tRNA
G A A
G A U
C U A C U U C G A
mRNA
75
aa1
peptide bond
aa3
aa2
1-tRNA
3-tRNA
U A C
(leaves)
2-tRNA
A U G
G A A
G A U
C U A C U U C G A
mRNA
Ribosomes move over one codon
76
aa1
peptide bonds
aa4
aa2
aa3
4-tRNA
2-tRNA
A U G
3-tRNA
G C U
G A U G A A
C U A C U U C G A A C U
mRNA
77
aa1
peptide bonds
aa4
aa2
aa3
2-tRNA
4-tRNA
G A U
(leaves)
3-tRNA
A U G
G C U
G A A
C U A C U U C G A A C U
mRNA
Ribosomes move over one codon
78
aa1
peptide bonds
aa5
aa2
aa3
aa4
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
79
peptide bonds
aa1
aa5
aa2
aa3
aa4
5-tRNA
U G A
3-tRNA
G A A
4-tRNA
G C U
G C U A C U U C G A A C U
mRNA
Ribosomes move over one codon
80
aa4
aa5
Termination
aa199
aa3 primary
structure
aa2 of a protein
aa200
aa1
200-tRNA
A C U
terminator
or stop
codon
C A U G U U U A G
mRNA
81
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
aa2
aa1
aa3
aa4
aa5
aa199
aa200
82
83
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
protein
codon 2
methionine
glycine
codon 3
serine
codon 4
isoleucine
codon 5
glycine
codon 6
alanine
codon 7
stop
codon
Primary structure of a protein
aa1
aa2
aa3
peptide bonds
aa4
aa5
aa6
84
Protein Synthesis
85