Lecture 22 - Molecular Basis of Genetics PDF
Lecture 22 - Molecular Basis of Genetics PDF
Lecture Presentation by
Nicole Tunbridge and
Kathleen Fitzpatrick
§ DNA is copied during DNA replication, and cells § Early in the 20th century, the identification of the
can repair their DNA molecules of inheritance loomed as a major
challenge to biologists
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The Search for the Genetic Material: Scientific Evidence That DNA Can Transform Bacteria
Inquiry
§ When T. H. Morgan’s group showed that genes § The discovery of the genetic role of DNA began
are located on chromosomes, the two components with research by Frederick Griffith in 1928
of chromosomes—DNA and protein—became
candidates for the genetic material § Griffith worked with two strains of a bacterium, one
pathogenic and one harmless
§ The role of DNA in heredity was first discovered
by studying bacteria and the viruses that
infect them
Figure 16.2
Experiment
Living S cells Living R cells Heat-killed S cells Mixture of heat-
(pathogenic (nonpathogenic (nonpathogenic killed S cells and
§ When he mixed heat-killed remains of the control) control) control) living R cells
Living S cells
§ In 1944, Oswald Avery, Maclyn McCarty, and § More evidence for DNA as the genetic material
Colin MacLeod announced that the transforming came from studies of viruses that infect bacteria
substance was DNA
§ Such viruses, called bacteriophages (or phages),
§ Many biologists remained skeptical, mainly are widely used in molecular genetics research
because little was known about DNA
§ A virus is DNA (sometimes RNA) enclosed by a
protective coat, often simply protein
2
Figure 16.3
Phage
head
DNA
Tail
sheath
Tail fiber
Genetic
material
Bacterial 100 nm
cell
Figure 16.4
Experiment
Batch 1: Radioactive sulfur (35S) in phage protein
1 Labeled phages 2 Agitation frees outside 3 Centrifuged cells
infect cells. phage parts from cells. form a pellet.
§ In 1952, Alfred Hershey and Martha Chase Radioactive 4 Radioactivity
(phage protein)
protein
showed that DNA is the genetic material of a found in liquid
phage known as T2
§ They designed an experiment showing that only
Centrifuge
one of the two components of T2 (DNA or protein)
Pellet
enters an E. coli cell during infection Batch 2: Radioactive phosphorus (32P) in phage DNA
Radioactive
DNA
§ They concluded that the injected DNA of the
phage provides the genetic information
Centrifuge
3
Figure 16.5
Sugar– Nitrogenous
phosphate
5′ end
bases
Animation: DNA and RNA Structure
backbone
Thymine (T)
Adenine (A)
Cytosine (C)
Phosphate
Guanine (G)
3′ end
Sugar
DNA (deoxyribose)
Nitrogenous
nucleotide base
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§ In any species the number of A and T bases are § Maurice Wilkins and Rosalind Franklin were using
equal and the number of G and C bases are equal a technique called X-ray crystallography to study
molecular structure
§ The basis for these rules was not understood until
the discovery of the double helix § Franklin produced a picture of the DNA molecule
using this technique
Figure 16.6
4
Figure 16.7
5′ end
C G
C G Hydrogen bond 3′ end
G C § Watson and Crick built models of a double helix to
G C T A
3.4 nm
conform to the X-rays and chemistry of DNA
T A
G
C
C
G
G C
§ Franklin had concluded that there were two outer
A T
sugar-phosphate backbones, with the nitrogenous
1 nm
T A
C G
bases paired in the molecule’s interior
C G
G C
C G A T § Watson built a model in which the backbones were
A T 3′ end antiparallel (their subunits run in opposite
T
A
A
T
0.34 nm
5′ end directions)
(a) Key features of (b) Partial chemical structure (c) Space-filling
DNA structure model
Figure 16.UN02
Figure 16.8
§ They determined that adenine (A) paired only with Sugar
thymine (T), and guanine (G) paired only with Adenine (A) Thymine (T)
cytosine (C)
§ The Watson-Crick model explains Chargaff’s
rules: in any organism the amount of A = T, and
the amount of G = C Sugar
Sugar
5
Concept 16.2: Many proteins work together in The Basic Principle: Base Pairing to a Template
DNA replication and repair Strand
§ The relationship between structure and function is § Since the two strands of DNA are complementary,
manifest in the double helix each strand acts as a template for building a new
strand in replication
§ Watson and Crick noted that the specific base
pairing suggested a possible copying mechanism § In DNA replication, the parent molecule unwinds,
for genetic material and two new daughter strands are built based on
base-pairing rules
A T A T A T
C G C G C G
T A T A T A
A T A T A T
G C G C G C
Figure 16.9-3
6
Figure 16.10
First Second
Parent cell replication replication
(a) Conservative
model
§ Experiments by Matthew Meselson and Franklin
Stahl supported the semiconservative model
§ They labeled the nucleotides of the old strands
(b) Semiconserva- with a heavy isotope of nitrogen, while any new
tive model
nucleotides were labeled with a lighter isotope
(c) Dispersive
model
Figure 16.11
Experiment
1 Bacteria cultured 2 Bacteria transferred
in medium with 15N to medium with 14N
(heavy isotope) (lighter isotope)
Conservative
model
Semiconservative
model
Dispersive
model
§ The copying of DNA is remarkable in its speed and § Replication begins at particular sites called origins
accuracy of replication, where the two DNA strands are
separated, opening up a replication “bubble”
§ More than a dozen enzymes and other proteins
participate in DNA replication § A eukaryotic chromosome may have hundreds or
even thousands of origins of replication
§ Replication proceeds in both directions from each
origin, until the entire molecule is copied
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Figure 16.12
(a) Origin of replication in an E. coli cell (b) Origins of replication in a eukaryotic cell
Origin of
Animation: Origins of Replication
Origin of Parental (template)
replication strand replication Eukaryotic chromosome
Daughter
(new) strand Parental (template)
Double-stranded strand
Replication DNA molecule Daughter (new)
Bacterial
fork strand
chromosome
Double-
Replication
stranded
bubble Replication
DNA molecule Bubble
fork
Two daughter
DNA molecules
0.25 µm
0.5 µm
Figure 16.13
Primase
§ DNA polymerases cannot initiate synthesis of a § An enzyme called primase can start an RNA
polynucleotide; they can only add nucleotides to chain from scratch and adds RNA nucleotides one
an existing 3′ end at a time using the parental DNA as a template
§ The initial nucleotide strand is a short RNA primer § The primer is short (5–10 nucleotides long), and
the 3′ end serves as the starting point for the new
DNA strand
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Synthesizing a New DNA Strand
§ Enzymes called DNA polymerases catalyze the § Each nucleotide that is added to a growing DNA
elongation of new DNA at a replication fork strand is a nucleoside triphosphate
§ Most DNA polymerases require a primer and a § dATP supplies adenine to DNA and is similar to
DNA template strand the ATP of energy metabolism
§ The rate of elongation is about 500 nucleotides per § The difference is in their sugars: dATP has
second in bacteria and 50 per second in human deoxyribose while ATP has ribose
cells
§ As each monomer of dATP joins the DNA strand, it
loses two phosphate groups as a molecule of
pyrophosphate
Figure 16.14
New strand Template strand
5′ 5′
Antiparallel Elongation
3′ 3′
Sugar A T A T
§ The antiparallel structure of the double helix
Phosphate Base affects replication
C G C G § DNA polymerases add nucleotides only to the free
3′ end of a growing strand; therefore, a new DNA
G C DNA G C
poly- strand can elongate only in the 5′ to 3′ direction
OH merase
3′ A T A
T OH
P P Pi
P C 3′ C
P Pyro-
OH phosphate
Nucleotide
5′ 5′
2 Pi
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.
Figure 16.15
Overview
Leading strand
Origin of replication
Lagging strand
Animation: Leading Strand
Primer
Leading strand
Lagging strand
Overall
directions
1 DNA pol III starts to of replication Origin of replication
synthesize leading
3′
strand.
5′
RNA primer
5′ 3′ Sliding clamp
3′
DNA pol III
Parental DNA 5′
3′
5′
5′ 2 Continuous
3′ 3′
elongation in the
5′ to 3′ direction
5′
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9
§ Along one template strand of DNA, the DNA § To elongate the other new strand, called the
polymerase synthesizes a leading strand lagging strand, DNA polymerase must work in
continuously, moving toward the replication fork the direction away from the replication fork
§ The lagging strand is synthesized as a series of
segments called Okazaki fragments, which are
joined together by DNA ligase
Figure 16.16
Overview
Lagging Origin of replication
strand
Animation: Lagging Strand
Leading Lagging
strand strand
2
1
Leading
strand RNA primer
Overall directions for fragment 2
of replication
5′ Okazaki 4 DNA pol III
1 Primase makes 3′ fragment 2 makes Okazaki
3′
RNA primer. 2 fragment 2.
Origin of
replication 3′
5′ 3′ 1
Template 5′ 3′ 5′
5′
strand 5′
2 DNA pol III 3′ 5 DNA pol I
RNA primer makes Okazaki replaces RNA
3′
for fragment 1 fragment 1. with DNA.
2
1 3′
5′
1 3′ 5′ 3′ 5′
5′
6 DNA ligase
5′ forms bonds
3 DNA pol III 3′ between DNA
3′ detaches. fragments.
Okazaki 2
fragment 1 1 3′
5′ 5′
1 3′
5′ Overall direction of replication
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Figure 16.17
Lagging strand 5′
template
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Table 16.1
Figure 16.18
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Figure 16.19-1 Figure 16.19-2
5′ 3′ 5′ 3′
3′ 5′ 3′ 5′
Nuclease Nuclease
5′ 3′ 5′ 3′
3′ 5′ 3′ 5′
DNA
polymerase
5′ 3′
3′ 5′
Figure 16.19-3
5′ 3′
Evolutionary Significance of Altered DNA
3′ 5′ Nucleotides
Nuclease
§ Error rate after proofreading repair is low but
not zero
5′ 3′
5′ 3′
3′ 5′
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Figure 16.20
5′
§ The usual replication machinery provides no way 3′
to complete the 5′ ends, so repeated rounds of Parental strand Removal of primers and
replacement with DNA
replication produce shorter DNA molecules with where a 3′ end is available
5′
uneven ends 3′
Second round
of replication
§ This is not a problem for prokaryotes, most of 5′
which have circular chromosomes New leading strand 3′
New lagging strand 5′
3′
Further rounds
of replication
Shorter and shorter daughter molecules
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12
Figure 16.21
§ If chromosomes of germ cells became shorter in § The shortening of telomeres might protect cells
every cell cycle, essential genes would eventually from cancerous growth by limiting the number of
be missing from the gametes they produce cell divisions
§ An enzyme called telomerase catalyzes the § There is evidence of telomerase activity in cancer
lengthening of telomeres in germ cells cells, which may allow cancer cells to persist
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Figure 16.22 Figure 16.22a
Chromatid
(700 nm)
Nucleosome
DNA
(10 nm in diameter)
30-nm fiber
Nucleosome
double helix (10 nm in diameter)
(2 nm in diameter)
DNA
H1 Loops Scaffold double helix
Histones
Histone tail 300-nm (2 nm in diameter)
fiber
DNA, the Histones Nucleosomes,
double helix or “beads on 30-nm fiber H1
Replicated
a string” Looped Histone tail
(10-nm fiber) domains
chromosome Histones
(1,400 nm)
(300-nm fiber)
Metaphase
chromosome DNA, the Histones Nucleosomes, or “beads on
double helix a string” (10-nm fiber)
Figure 16.23
5 µm
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14
§ Most chromatin is loosely packed in the nucleus § Histones can undergo chemical modifications that
during interphase and condenses prior to mitosis result in changes in chromatin organization
§ Loosely packed chromatin is called euchromatin
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