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![Page 1: Chapter 16- molecular biology presentation](https://reader033.fdocuments.in/reader033/viewer/2022052514/577cc9bc1a28aba711a478c9/html5/thumbnails/1.jpg)
LECTURE PRESENTATIONSFor CAMPBELL BIOLOGY, NINTH EDITION
Jane B. Reece, Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Robert B. Jackson
© 2011 Pearson Education, Inc.
Lectures byErin Barley
Kathleen Fitzpatrick
The Molecular Basis of Inheritance
Chapter 16
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Figure 16.1
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Concept 16.1: DNA is the genetic material
• Early in the 20th century, the identification of the molecules of inheritance loomed as a major challenge to biologists
© 2011 Pearson Education, Inc.
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The Search for the Genetic Material: Scientific Inquiry
• When T. H. Morgan’s group showed that genes are located on chromosomes, the two components of chromosomes—DNA and protein—became candidates for the genetic material
• The key factor in determining the genetic material was choosing appropriate experimental organisms
• The role of DNA in heredity was first discovered by studying bacteria and the viruses that infect them
© 2011 Pearson Education, Inc.
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Living S cells(control)
Living R cells(control)
Heat-killedS cells(control)
Mixture ofheat-killedS cells andliving R cells
Mouse dies Mouse diesMouse healthy Mouse healthy
Living S cells
EXPERIMENT
RESULTS
Figure 16.2
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Figure 16.3
Phagehead
Tailsheath
Tail fiber
DNA
Bacterialcell
100
nm
Evidence That Viral DNA Can Program Cells
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Figure 16.4-1
Bacterial cell
Phage
Batch 1:Radioactivesulfur(35S)
DNA
Batch 2:Radioactivephosphorus(32P)
RadioactiveDNA
EXPERIMENTRadioactiveprotein
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Figure 16.4-2
Bacterial cell
Phage
Batch 1:Radioactivesulfur(35S)
Radioactiveprotein
DNA
Batch 2:Radioactivephosphorus(32P)
RadioactiveDNA
Emptyproteinshell
PhageDNA
EXPERIMENT
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Figure 16.4-3
Bacterial cell
Phage
Batch 1:Radioactivesulfur(35S)
Radioactiveprotein
DNA
Batch 2:Radioactivephosphorus(32P)
RadioactiveDNA
Emptyproteinshell
PhageDNA
Centrifuge
Centrifuge
Radioactivity(phage protein)in liquid
Pellet (bacterialcells and contents)
PelletRadioactivity(phage DNA)in pellet
EXPERIMENT
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© 2011 Pearson Education, Inc.
Animation: Hershey-Chase ExperimentRight-click slide / select “Play”
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Additional Evidence That DNA Is the Genetic Material
• It was known that DNA is a polymer of nucleotides, each consisting of a nitrogenous base, a sugar, and a phosphate group
• In 1950, Erwin Chargaff reported that DNA composition varies from one species to the next
• This evidence of diversity made DNA a more credible candidate for the genetic material
© 2011 Pearson Education, Inc.
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© 2011 Pearson Education, Inc.
Animation: DNA and RNA Structure Right-click slide / select “Play”
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• Two findings became known as Chargaff’s rules– The base composition of DNA varies between
species– In any species the number of A and T bases are
equal and the number of G and C bases are equal• The basis for these rules was not understood until
the discovery of the double helix
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Figure 16.5 Sugar–phosphatebackbone
Nitrogenous bases
Thymine (T)
Adenine (A)
Cytosine (C)
Guanine (G)
Nitrogenous base
Phosphate
DNA nucleotide
Sugar(deoxyribose)
3 end
5 end
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Building a Structural Model of DNA: Scientific Inquiry
• After DNA was accepted as the genetic material, the challenge was to determine how its structure accounts for its role in heredity
• Maurice Wilkins and Rosalind Franklin were using a technique called X-ray crystallography to study molecular structure
• Franklin produced a picture of the DNA molecule using this technique
© 2011 Pearson Education, Inc.
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Figure 16.6
(a) Rosalind Franklin (b) Franklin’s X-ray diffractionphotograph of DNA
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• Franklin’s X-ray crystallographic images of DNA enabled Watson to deduce that DNA was helical
• The X-ray images also enabled Watson to deduce the width of the helix and the spacing of the nitrogenous bases
• The pattern in the photo suggested that the DNA molecule was made up of two strands, forming a double helix
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© 2011 Pearson Education, Inc.
Animation: DNA Double Helix Right-click slide / select “Play”
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• Watson and Crick built models of a double helix to conform to the X-rays and chemistry of DNA
• Franklin had concluded that there were two outer sugar-phosphate backbones, with the nitrogenous bases paired in the molecule’s interior
• Watson built a model in which the backbones were antiparallel (their subunits run in opposite directions)
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3.4 nm
1 nm
0.34 nm
Hydrogen bond
(a) Key features ofDNA structure
(b) Partial chemical structure
3 end
5 end
3 end
5 end
T
T
A
A
G
G
C
C
CC
CC
C
C
CC
C
GG
GG
G
G
GG
G
T
T
T
T
TT
A
A
A
A
AA
Figure 16.7a
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• At first, Watson and Crick thought the bases paired like with like (A with A, and so on), but such pairings did not result in a uniform width
• Instead, pairing a purine with a pyrimidine resulted in a uniform width consistent with the X-ray data
© 2011 Pearson Education, Inc.
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Figure 16.UN01
Purine purine: too wide
Pyrimidine pyrimidine: too narrow
Purine pyrimidine: widthconsistent with X-ray data
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• Watson and Crick reasoned that the pairing was more specific, dictated by the base structures
• They determined that adenine (A) paired only with thymine (T), and guanine (G) paired only with cytosine (C)
• The Watson-Crick model explains Chargaff’s rules: in any organism the amount of A = T, and the amount of G = C
© 2011 Pearson Education, Inc.
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Figure 16.8
Sugar
Sugar
Sugar
Sugar
Adenine (A) Thymine (T)
Guanine (G) Cytosine (C)
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Concept 16.2: Many proteins work together in DNA replication and repair
• The relationship between structure and function is manifest in the double helix
• Watson and Crick noted that the specific base pairing suggested a possible copying mechanism for genetic material
© 2011 Pearson Education, Inc.
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The Basic Principle: Base Pairing to a Template Strand
• Since the two strands of DNA are complementary, each strand acts as a template for building a new strand in replication
• In DNA replication, the parent molecule unwinds, and two new daughter strands are built based on base-pairing rules
© 2011 Pearson Education, Inc.
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© 2011 Pearson Education, Inc.
Animation: DNA Replication OverviewRight-click slide / select “Play”
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Figure 16.9-1
(a) Parent molecule
A
A
A
T
T
T
C
C
G
G
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Figure 16.9-2
(a) Parent molecule (b) Separation ofstrands
A
A
A
A
A
A
T
T
T
T
T
T
C
C
C
C
G
G
G
G
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Figure 16.9-3
(a) Parent molecule (b) Separation ofstrands
(c)“Daughter” DNA molecules,each consisting of oneparental strand and onenew strand
A
A
A
A
A
A
A
A
A
A
A
A
T
T
T
T
T
T
T
T
T
T
T
T
C
C
C
C
C
C
C
C
G
G
G
G
G
G
G
G
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Figure 16.10
(a) Conservativemodel
(b) Semiconservativemodel
(c) Dispersive model
Parentcell
Firstreplication
Secondreplication
Two parent strands rejoin
each strand is a mix of old and new
One old and one new strand
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Figure 16.11a
Bacteriacultured inmedium with15N (heavyisotope)
Bacteriatransferred tomedium with14N (lighterisotope)
DNA samplecentrifugedafter firstreplication
DNA samplecentrifugedafter secondreplication
Less dense
More dense
21
3 4
EXPERIMENT
RESULTS
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Figure 16.11b
Predictions: First replication Second replication
Conservativemodel
Semiconservativemodel
Dispersivemodel
CONCLUSION
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DNA Replication: A Closer Look
• The copying of DNA is remarkable in its speed and accuracy
• More than a dozen enzymes and other proteins participate in DNA replication
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© 2011 Pearson Education, Inc.
Animation: Origins of ReplicationRight-click slide / select “Play”
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Figure 16.12b
(b) Origins of replication in a eukaryotic cell
Origin of replicationDouble-strandedDNA molecule
Parental (template)strand
Daughter (new)strand
Bubble Replication fork
Two daughter DNA molecules0.25 m
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Figure 16.13
Topoisomerase
Primase
RNAprimer
Helicase
Single-strand bindingproteins
5
3
5
53
3
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• At the end of each replication bubble is a replication fork, a Y-shaped region where new DNA strands are elongating
• Helicases are enzymes that untwist the double helix at the replication forks
• Single-strand binding proteins bind to and stabilize single-stranded DNA
• Topoisomerase corrects “overwinding” ahead of replication forks by breaking, swiveling, and rejoining DNA strands
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Synthesizing a New DNA Strand
• Enzymes called DNA polymerases catalyze the elongation of new DNA at a replication fork
• Most DNA polymerases require a primer and a DNA template strand
• The rate of elongation is about 500 nucleotides per second in bacteria and 50 per second in human cells
© 2011 Pearson Education, Inc.
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Figure 16.14
New strand Template strand
Sugar
Phosphate Base
Nucleosidetriphosphate
DNApolymerase
Pyrophosphate
5
5
5
5
3
3
3
3
OH
OH
OH
P P i
2 P i
PP
P
A
A
A
A
T T
TT
C
C
C
C
C
C
G
G
G
G
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Antiparallel Elongation
• The antiparallel structure of the double helix affects replication
• DNA polymerases add nucleotides only to the free 3end of a growing strand; therefore, a new DNA strand can elongate only in the 5to3direction
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Figure 16.15
Leadingstrand
Laggingstrand
Overview
Origin of replication Laggingstrand
Leadingstrand
Primer
Overall directionsof replication
Origin of replication
RNA primer
Sliding clamp
DNA pol IIIParental DNA
35
5
33
5
3
5
3
5
3
5
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© 2011 Pearson Education, Inc.
Animation: Leading Strand Right-click slide / select “Play”
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Figure 16.15a
Leadingstrand
Laggingstrand
Overview
Origin of replication Laggingstrand
Leadingstrand
Primer
Overall directionsof replication
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© 2011 Pearson Education, Inc.
Animation: Lagging Strand Right-click slide / select “Play”
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Origin of replication
Overview
Leadingstrand
Leadingstrand
Laggingstrand
Lagging strand
Overall directionsof replication
Templatestrand
RNA primerfor fragment 1
Okazakifragment 1
RNA primerfor fragment 2
Okazakifragment 2
Overall direction of replication
3
3
3
3
3
3
3
3
3
3
3
3
5
5
5
5
5
55
55
55
5
2
2
21
1
1
1
12
1
Figure 16.16
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Figure 16.17
OverviewLeadingstrand
Origin of replication Lagging
strand
LeadingstrandLagging
strand Overall directionsof replicationLeading strand
DNA pol III
DNA pol III Lagging strandDNA pol I DNA ligase
PrimerPrimase
ParentalDNA
5
5
5
5
5
33
3
333 2 1
4
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The DNA Replication Complex
• The proteins that participate in DNA replication form a large complex, a “DNA replication machine”
• The DNA replication machine may be stationary during the replication process
• Recent studies support a model in which DNA polymerase molecules “reel in” parental DNA and “extrude” newly made daughter DNA molecules
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Animation: DNA Replication ReviewRight-click slide / select “Play”
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Figure 16.18
Parental DNADNA pol III
Leading strand
Connectingprotein
Helicase
Lagging strandDNA pol III
Laggingstrandtemplate
5
5
5
5
5
5
3 3
33
3
3
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Proofreading and Repairing DNA
• DNA polymerases proofread newly made DNA, replacing any incorrect nucleotides
• In mismatch repair of DNA, repair enzymes correct errors in base pairing
• DNA can be damaged by exposure to harmful chemical or physical agents such as cigarette smoke and X-rays; it can also undergo spontaneous changes
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Figure 16.19
Nuclease
DNA polymerase
DNA ligase
5
5
5
5
5
5
5
5
3
3
3
3
3
3
3
3
In nucleotide excision repair, a nuclease cuts out and replaces damaged stretches of DNA
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Evolutionary Significance of Altered DNA Nucleotides
• Error rate after proofreading repair is low but not zero
• Sequence changes may become permanent and can be passed on to the next generation
• These changes (mutations) are the source of the genetic variation upon which natural selection operates
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Replicating the Ends of DNA Molecules
• Limitations of DNA polymerase create problems for the linear DNA of eukaryotic chromosomes
• The usual replication machinery provides no way to complete the 5 ends, so repeated rounds of replication produce shorter DNA molecules with uneven ends
• This is not a problem for prokaryotes, most of which have circular chromosomes
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Figure 16.20
Ends of parentalDNA strands
Leading strandLagging strand
Last fragment Next-to-last fragment
Lagging strand RNA primer
Parental strand Removal of primers andreplacement with DNAwhere a 3 end is available
Second roundof replication
Further roundsof replication
New leading strandNew lagging strand
Shorter and shorter daughter molecules
3
3
3
3
3
5
5
5
5
5
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• If chromosomes of germ cells became shorter in every cell cycle, essential genes would eventually be missing from the gametes they produce
• An enzyme called telomerase catalyzes the lengthening of telomeres in germ cells
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• Eukaryotic chromosomal DNA molecules have special nucleotide sequences at their ends called telomeres
• Telomeres do not prevent the shortening of DNA molecules, but they do postpone the erosion of genes near the ends of DNA molecules
• It has been proposed that the shortening of telomeres is connected to aging
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• The shortening of telomeres might protect cells from cancerous growth by limiting the number of cell divisions
• There is evidence of telomerase activity in cancer cells, which may allow cancer cells to persist
© 2011 Pearson Education, Inc.