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Lecture 22 - Molecular Basis of Genetics PDF

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Lecture 22 - Molecular Basis of Genetics PDF

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Rosemond Fabien
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CAMPBELL

Life’s Operating Instructions


BIOLOGY TENTH
EDITION

Reece • Urry • Cain • Wasserman • Minorsky • Jackson


§  In 1953, James Watson and Francis Crick
introduced an elegant double-helical model for the

16 structure of deoxyribonucleic acid, or DNA


§  Hereditary information is encoded in DNA and
The Molecular reproduced in all cells of the body
Basis of §  This DNA program directs the development of
Inheritance biochemical, anatomical, physiological, and
(to some extent) behavioral traits

Lecture Presentation by
Nicole Tunbridge and
Kathleen Fitzpatrick

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.1 Figure 16.1a

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Concept 16.1: DNA is the genetic material

§  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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

1
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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

pathogenic strain with living cells of the harmless


strain, some living cells became pathogenic
§  He called this phenomenon transformation, now Results
defined as a change in genotype and phenotype
Mouse dies Mouse healthy Mouse healthy Mouse dies
due to assimilation of foreign DNA

Living S cells

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Evidence That Viral DNA Can Program 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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

2
Figure 16.3

Animation: Phage T2 Reproductive Cycle

Phage
head

DNA

Tail
sheath
Tail fiber
Genetic
material

Bacterial 100 nm
cell

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

Pellet 4 Radioactivity (phage


© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.
DNA) found in pellet

Animation: Hershey-Chase Experiment 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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Building a Structural Model of DNA: Scientific


Inquiry
§  Two findings became known as Chargaff’s rules §  After DNA was accepted as the genetic material,
the challenge was to determine how its structure
§  The base composition of DNA varies between
accounts for its role in heredity
species

§  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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.6

§  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
(a) Rosalind Franklin (b) Franklin’s X-ray diffraction double helix
photograph of DNA

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.UN02

§  At first, Watson and Crick thought the bases


paired like with like (A with A, and so on), but such Purine + purine: too wide
pairings did not result in a uniform width
§  Instead, pairing a purine with a pyrimidine resulted
Pyrimidine + pyrimidine: too narrow
in a uniform width consistent with the X-ray data

Purine + pyrimidine: width


consistent with X-ray data

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.8

§  Watson and Crick reasoned that the pairing was


more specific, dictated by the base structures Sugar

§  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

Guanine (G) Cytosine (C)


© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.9-1 Figure 16.9-2

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

(a) Parental (a) Parental (b) Separation of parental


molecule molecule strands into templates

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.9-3

§  Watson and Crick’s semiconservative model of


A T A T A T A T
replication predicts that when a double helix
C G C G C G C G
replicates, each daughter molecule will have one
T A T A T A T A old strand (derived or “conserved” from the parent
A T A T A T A T molecule) and one newly made strand
G C G C G C G C
§  Competing models were the conservative model
(a) Parental (b) Separation of parental (c) Formation of new strands (the two parent strands rejoin) and the dispersive
molecule strands into templates complementary to template model (each strand is a mix of old and new)
strands

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.11
Experiment
1 Bacteria cultured 2 Bacteria transferred
in medium with 15N to medium with 14N
(heavy isotope) (lighter isotope)

§  The first replication produced a band of hybrid


DNA, eliminating the conservative model Results
3 DNA sample 4 DNA sample Less dense
centrifuged centrifuged
§  A second replication produced both light and after first
replication
after second
replication More dense
hybrid DNA, eliminating the dispersive model and Conclusion
supporting the semiconservative model Predictions: First replication Second replication

Conservative
model

Semiconservative
model

Dispersive
model

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

DNA Replication: A Closer Look Getting Started

§  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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

7
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

Two daughter DNA molecules

0.25 µm
0.5 µm

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.13

Primase

§  At the end of each replication bubble is a Topoisomerase


replication fork, a Y-shaped region where 3′
new DNA strands are elongating 5′
RNA
3′ primer
§  Helicases are enzymes that untwist the double 5′
Replication
helix at the replication forks 3′ fork

§  Single-strand binding proteins bind to and


stabilize single-stranded DNA 5′
Helicase
§  Topoisomerase corrects “overwinding” ahead of Single-strand binding
replication forks by breaking, swiveling, and proteins
rejoining DNA strands
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

§  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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

8
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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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′
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.17

Animation: DNA Replication Overview


Overview
Leading Origin of
strand replication
Lagging
Leading strand strand
template 3′
Leading
Single-strand Lagging strand
strand 5′
binding proteins
Leading strand Overall directions
Helicase of replication
5′ DNA pol III

3′ Primer
3′ 5′ Primase
3′
Parental 5 DNA pol III
Lagging
DNA 5′ DNA pol I

4 3′ strand DNA ligase
5′
3 2 1
3′

Lagging strand 5′
template

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

10
Table 16.1

Animation: DNA Replication Review

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.18

The DNA Replication Complex Leading strand template

DNA pol III



§  The proteins that participate in DNA replication Leading strand
Parental DNA
form a large complex, a “DNA replication 5′
5′ 3′ 3′
machine”
3′ 3′
5′ 5′
§  The DNA replication machine may be stationary Connecting protein
during the replication process DNA pol III

Helicase

§  Recent studies support a model in which DNA Lagging 3′ 5′


polymerase molecules “reel in” parental DNA and strand
5′ 3′ Lagging strand
template
“extrude” newly made daughter DNA molecules

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

BioFlix: DNA Replication 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

§  In nucleotide excision repair, a nuclease cuts


out and replaces damaged stretches of DNA
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

11
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′

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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′

3′ 5′ §  Sequence changes may become permanent and


DNA can be passed on to the next generation
polymerase
§  These changes (mutations) are the source of the
5′ 3′
genetic variation upon which natural selection
3′ 5′ operates
DNA
ligase

5′ 3′

3′ 5′
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.20

Replicating the Ends of DNA Molecules Ends of parental


5′
Leading strand
DNA strands Lagging strand
3′

§  Limitations of DNA polymerase create problems Last fragment


Next-to-last
fragment
for the linear DNA of eukaryotic chromosomes Lagging strand RNA primer

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
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

12
Figure 16.21

§  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 1 µm

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

§  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

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Concept 16.3: A chromosome consists of a DNA


molecule packed together with proteins
§  The bacterial chromosome is a double-stranded, §  In the eukaryotic cell, DNA is precisely combined
circular DNA molecule associated with a small with proteins in a complex called chromatin
amount of protein
§  Chromosomes fit into the nucleus through an
§  Eukaryotic chromosomes have linear DNA elaborate, multilevel system of packing
molecules associated with a large amount
of protein

§  In a bacterium, the DNA is “supercoiled” and


found in a region of the cell called the nucleoid

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

13
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)

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Animation: DNA Packing Video: Cartoon and Stick Model of a


Nucleosomal Particle

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

Figure 16.23

§  Chromatin undergoes changes in packing during


the cell cycle
§  At interphase, some chromatin is organized into a
10-nm fiber, but much is compacted into a 30-nm
fiber, through folding and looping
§  Interphase chromosomes occupy specific
restricted regions in the nucleus and the fibers of
different chromosomes do not become entangled

5 µm
© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

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

§  During interphase a few regions of chromatin


(centromeres and telomeres) are highly
condensed into heterochromatin

§  Dense packing of the heterochromatin makes it


difficult for the cell to express genetic information
coded in these regions

© 2014 Pearson Education, Inc. © 2014 Pearson Education, Inc.

15

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