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Page 1: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

LECTURE 1

Introduction to DNA and RNA

(Chapter 09)

1

Colored background: Review on your own

Page 2: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

A BRIEF INTRODUCTION TO LIFE

• Genetics: Study of the structure, function, and transmission of genes

• Only living organisms have genes• To understand genetics, we will start with the

question: “What is Life?”– Characteristics shared by all living forms but not

by non-living forms

2

Group Exercise

Page 3: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

What is the role of genes in life?

• DNA carries information to create order• Life “got going” as a negative entropy machine• Why? Because it did! • It’s a marvel!

3

HIGH ENTROPY (DISORDER)

Arrow of time

LOW ENTROPY(ORDER)

Page 4: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

4

Time

22 deg C(surrounding

air)

100º C 22º C

22.5 deg C(surrounding

air)

ENERGY

Page 5: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

5

HIGH ENTROPY (DISORDER)

Arrow of time

LOW ENTROPY(ORDER)

Can’t turn back

time so use

ENERGY instead!

Page 6: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Where does the energy come from?

6

High potential energy; Low entropy

Short mean λ

Longer mean λ

Life!

Page 7: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• Fusion reactions in sun’s core– Hydrogen to helium with release of photons

– Mass converted to energy (E = mc2)– 600 million tons hydrogen fused to 596 million tons helium

per second– 4 million tons hydrogen converted to 3.9 x 1026 Joules

7

4 H 1 He

Page 8: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Fusion fueled by GRAVITY

8

It’s why the sun shines!

Page 9: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• The source of all life’s “problems” is ENTROPY• The “solutions” are stored as genes in DNA

9

DnA iS DiGiTal Information

A,C,G,T

Page 10: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

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• DNA is a highly ORDERED molecule that…• Fights entropy to perpetuate itself• By…Building its own survival machines

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12

• And…passing them on• Geneticists view this as…

• a “blind” undirected process

Page 13: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

13

Life = Problems

“I had trouble in getting to Solla Sollew where they never have troubles at least very few…” –Dr. Suess

Page 14: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

14

There is no happy retirement from life. Forget it!

I hope we can get little

Davey off meth…

I hate my thighs…

Page 15: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

15

Practice Problem:(Identify all correct answers)

From a geneticist’s point-of-view, which of the following statements is true?a. Life is trying to evolveb. A bee is a survival machine for its genesc. DNA is a digital information moleculed. Life requires an energy source to fight entropye. When you graduate from college, all your problems

will disappear

Page 16: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

MOLECULAR GENETICS

• The study of the structure and function of DNA at the molecular level– Highly ordered digital information– Survives to perpetuate itself

• “Selfish gene”

16

Page 17: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

To do this DNA must…

• 1. Carry the information necessary to encode its survival machine– Reverse entropy– Perpetuate itself

• 2. Be able to transmit this information to daughter cells– Reproduction

• 3. Be capable of change– Subject to mutation

17

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18

Could life exist if DNA did not have these three properties?

What would happen to this primitive cell if its DNA did not encode its structures/functions, it could not reproduce, and/or it’s DNA was not capable of change?

Page 19: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

IDENTIFICATION OF DNA AS THE GENETIC MATERIAL

• Early humans noticed that traits were inherited between parents and offspring

19

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• The substance that held this “heritable information” was mysterious

• Mendel’s work in mid-1800s revealed patterns (“laws”) that govern its inheritance

• In 1901, Hugo de Vries and Carlos Correns linked these laws to chromosome behavior during meiosis

20

We’ll study Mendel’s Laws and their connection to meiosis later on this semester

Page 21: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• Early biochemists studied the chemical components in the nucleus to try to identify the heritable information molecule

• Nucleus contains proteins and nucleic acids– Proteins are more chemically diverse than nucleic

acids– Known to do almost everything in the cell– Most biologists hypothesized that the heritable

material was a protein

• Starting in 1928, three seminal experiments shattered this hypothesis– Revealed DNA as the heritable molecule

21

Page 22: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

1. Frederick Griffith Experiments with Streptococcus pneumonia (1928)

• Griffith studied a bacterium (pneumococci) now known as Streptococcus pneumoniae

• S. pneumoniae comes in two strains– S Smooth

• Secrete a polysaccharide capsule– Protects bacterium from the immune system

of animals

• Produce smooth colonies on solid media– R Rough

• Unable to secrete a capsule• Produce colonies with a rough

appearance22

Page 23: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• Experimental tools:– R cells– S cells– Mice– Agar plating

23

Page 24: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.1

Living type S bacteria wereinjected into a mouse.

Mouse died

Deadtype S

Livetype R

Mouse died Mouse survived Mouse survived

Living type R bacteria wereinjected into a mouse.

Heat-killed type S bacteriawere injected into a mouse.

Living type R and heat-killedtype S bacteria were injectedinto a mouse.

Type S bacteria were isolatedfrom the dead mouse.

No living bacteria were isolatedfrom the mouse.

No living bacteria were isolatedfrom the mouse.

Type S bacteria were isolatedfrom the dead mouse.

(a) Live type S (b) Live type R (c) Dead type S (d) Live type R + dead type S

Afterseveraldays

Afterseveraldays

Afterseveraldays

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Afterseveraldays

24

Page 25: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Griffith concluded that something from the dead type S bacteria was transforming type R bacteria into type S Change was stable and heritable He called this process transformation

Provided an experimental model for studying the molecule that carries heritable information Griffith did not know what it was His experiment suggested that it was resistant to heat

First experimental indication that the heritable molecule was not a protein

Most proteins are sensitive to heat

25

We will be transforming bacteria in lab

Page 26: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

As an aside… We now know that the formation of the

capsule is guided by the bacteria’s DNA Transformed R bacteria acquired a gene from

the dead lysed S bacteria that directs the cell how to make a polysachharide capsule

Variation exists in ability to make capsule (R cells carry a non-functional allele for the gene)

The gene required to create a capsule is replicated and transmitted from mother to daughter cells

Once R cells acquired it they passed the trait to their “offspring”

26

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27

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28

Practice Problem(Only one correct answer):

If Griffith had mixed heat-killed R cells with living S cells and injected the mixture into his mice, what would have happened?a. Mice alive; only smooth colonies on plateb. Mice dead; only smooth colonies on platec. Mice alive; only rough colonies on plated. Mice dead; only rough colonies on platee. Mice dead; Both smooth and rough colonies on plate

Page 29: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

2. The Experiments of Avery, MacLeod and McCarty (1944)

• Avery, MacLeod and McCarty realized that Griffith’s observations could be used to identify the genetic material

• They carried out their experiments in the 1940s– At that time, it was known that DNA, RNA, proteins and

carbohydrates are the major constituents of living cells

• They prepared cell extracts from type S cells and purified each type of macromolecule

• Then repeated Griffith’s experiment without the mice!

29

X

Page 30: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• Replaced the mice with an antibody that binds to R cells

• Served the same purpose as the mouse’s immune system– Rather than killing the cells it allowed them to pull them out of

the cell mixtures prior to plating

• Experimental tools:– R cells– S cells– Antibody to R cells– DNase– Rnase– Proteinase– Agar plating

30

Page 31: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.2 Avery et al. conducted the following experiments

To further verify that DNA, and not a contaminant (RNA or protein), is the genetic material

Mix Mix Mix Mix

Type Rcells

Type Rcells

Type SDNA

extract

Type Rcells

Type SDNA

extract+

DNase

Type Rcells

Type SDNA

extract+

RNase

Type Rcells

Type SDNA

extract+

protease

Transformed Transformed Transformed

1 2 3 4 5

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Allow sufficient time for the DNA to be taken up by the type R bacteria. Only a small percentage of the type R bacteria will be transformed to type S.

Add an antibody that aggregates type R bacteria (that have not been transformed). The aggregated bacteria are removed by gentle centrifugation.

Plate the remaining bacteria on petri plates. Incubate overnight.

31

Page 32: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

32

Practice Problem(Circle all correct answers):

Why did Avery, McLeod, and McCarty use an antibody to R cells in their experiment?a. To protect their mice from dyingb. So they could detect smooth colonies if they were presentc. So they could repeat the essential features of Griffith’s

experiment without having to sacrifice animalsd. Because only R cells have a polysaccharide capsulee. Because only S cells have a polysaccharide capsule

Page 33: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

3. Hershey and Chase Experiment with Bacteriophage T2 (1952)

• Alfred Hershey and Marsha Chase provided further evidence that DNA is the genetic material

33

Page 34: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• Experimental tools:

– Bacteriophage T2– S35 (labels proteins)– P32 (labels DNA)– E. coli– A Waring brand kitchen blender

Head

Tail fiber

Base plate

Sheath

DNA(inside thecapsid head)

Made up of protein

Inside the

capsid

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

34

Infects E. coli cells by binding to the outside and injecting their DNA

Page 35: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

35

Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.

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Page 38: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

80%Blending removes 80%of 35S from E. coli cells.

35%Most of the 32P (65%)remains with intactE. coli cells.

Tota

l iso

top

e in

su

per

nat

ant

(%)

Agitation time in blender (min)

100

80

60

40

20

00 1 2 3 4 5 6 7 8

Data from A. D. Hershey and Martha Chase (1952) Independent Functions of Viral Protein and Nucleic Acid in Growth ofBacteriophage. Journal of General Physiology 36, 39–56.

Extracellular 35S

Extracellular 32P

The Data Most of the 35S was found in the supernatant

But only a small percentage of 32P

These results suggest that DNA is injected into the bacterial cytoplasm during infection

Data is not conclusive since less than 100% of the DNA or protein ended up in the cell or supernatant

Data is consistent with the hypothesis that DNA is the genetic material38

Page 39: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

39

Practice Problem(Circle all correct answers):

Which of the following were used in the Hershey-Chase experiment?a. E. colib. Bacteriophage T2c. S cellsd. A kitchen blendere. 35S

Page 40: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

In 1956, A. Gierer and G. Schramm isolated RNA from the tobacco mosaic virus (TMV), a plant virus Purified RNA caused the same lesions as intact TMV

viruses Therefore, the viral genome is composed of RNA

Since that time, many RNA viruses have been found Refer to Table 9.1

RNA Functions as the Genetic Material in Some Viruses

40

Page 41: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

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Page 42: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

9.2 NUCLEIC ACID STRUCTURE – On Your Own

• DNA and RNA are large macromolecules with several levels of complexity

– 1. Nucleotides form the repeating unit of nucleic acids– 2. Nucleotides are linked to form a linear strand of RNA

or DNA– 3. Two strands can interact to form a double helix– 4. The 3-D structure of DNA results from folding and

bending of the double helix. Interaction of DNA with proteins produces chromosomes within living cells

– Refer to fig. 9.6

42

Page 43: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.6

CC

G

AA

GA

TG

GT

T TA

A

GC

AT

AT

T TA

GC

Nucleotides

Single strand

Double helix

Three-dimensional structure

A T CG

CA A

T CA TG

CA

AT C

GC

A

G CA T

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

43

Page 44: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

The nucleotide is the repeating structural unit of DNA and RNA

It has three components A phosphate group A pentose sugar A nitrogenous base

Refer to Figure 9.7

Nucleotides

44

Page 45: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

9-24Figure 9.7

Phosphate group Sugars

D-Deoxyribose (in DNA)

Purines(double ring)

Pyrimidines(single ring)

Bases

OO

O–

O–

P

H

H

H

HO

OHOHOCH2

HH

D-Ribose (in RNA)

H

OH

H

HO

OHOHOCH2

HH

Uracil (U) (in RNA)Thymine (T) (in DNA)

Cytosine (C)

Adenine (A)

Guanine (G)

NH2

N

HH

H

H

H

O

N

43

2

156

7

89

43

21

5

6

O

CH3 H

43

2

156

7

8

9

5′

O

NH2

H

HN

N

N

N

NH2

N

N

H

N

N

N

H

H O

N43

21

5

6

O

H

H O

43

21

5

6

N

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

4′ 1′

2′3′

5′

4′ 1′

3′ 2′

N

45

Page 46: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

HH

H

OCH2

Base

(a) Repeating unit of deoxyribonucleic acid (DNA)

(b) Repeating unit of ribonucleic acid (RNA)

Phosphate

Deoxyribose

HH

OH

OCH2

Base

Phosphate

Ribose

5′

OH OH

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

4′ 1′

3′ 2′

5′

4′ 1′

3′ 2′

OO

O

P

O–

O

O

P

O–

O

HH HH

Figure 9.8 The structure of nucleotides found in (a) DNA and (b) RNA

A, G, C or T

These atoms are found within individual nucleotides However, they are removed when nucleotides join together to make

strands of DNA or RNA

A, G, C or U

46

Page 47: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Copyright ©The McGraw-Hill Companies, Inc. Permission required for reproduction or display

Base + sugar nucleoside Example

Adenine + ribose = Adenosine Adenine + deoxyribose = Deoxyadenosine

Base + sugar + phosphate(s) nucleotide Example

Adenosine monophosphate (AMP) Adenosine diphosphate (ADP) Adenosine triphosphate (ATP)

Refer to Figure 9.9

47

Page 48: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.9

Base always attached here

Phosphates are attached here

Adenosine

Adenosine monophosphate

Adenosine diphosphate

Adenine

Phosphate groups

Phosphoester bond

Ribose

H

OP CH2

O–

OO P

O–

O O O

–O P

O–

H

OHHO

O

H

2′3

1′4′5′

Adenosine triphosphate

NH2

N

H

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

H

N

NN

48

Page 49: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Nucleotides are covalently linked together by phosphodiester bonds A phosphate connects the 5’ carbon of one nucleotide to

the 3’ carbon of another Therefore the strand has directionality

5’ to 3’ In a strand, all sugar molecules are oriented in the same

direction

The phosphates and sugar molecules form the backbone of the nucleic acid strand The bases project from the backbone

49

Page 50: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.10

NH2

N O

N

O

N O

N

Adenine (A)

Guanine (G)

Thymine (T)

BasesBackbone

Cytosine (C)

O

HH

H

H

HH

OOO

O–

P CH2

O–

HH

H

H

H

HH

OOO

O

P CH2

O–

NH2

N

N

H

N

N

HH

H

HH

OOO

O

P CH2

O–

NH2

HN

N

N

H

N

HH

HOH

HH

OOO

O

P CH2

O–Singlenucleotide

Phosphodiesterlinkage

Sugar (deoxyribose)

Phosphate

3′

5′

5′4′ 1′

2′3′

5′4′ 1′

2′3′

5′4′ 1′

2′3′

5′4′ 1′

2′3′

CH3

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

50

Page 51: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

In 1953, James Watson and Francis Crick elucidated the double helical structure of DNA

The scientific framework for their breakthrough was provided by other scientists including Linus Pauling Rosalind Franklin and Maurice Wilkins Erwin Chargaff

Key Events Leading to the Discovery of the Double Helix

51

Page 52: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

In the early 1950s, he proposed that regions of protein can fold into a secondary structure a-helix

Linus Pauling

To elucidate this structure, he built ball-and-stick models

52

Page 53: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

53

Figure 9.11

CC

C

CCC

O

C

O

OC

OH

H

H

H

N

CN

NN

CC

CC O

O

HHNN

C

C

C

CCC

OC

O

C

O

OC

OH

HH

N

NN

H

N

H

N

Carbonyl oxygenAmide hydrogen

Hydrogen bond

(a) An a helix in a protein

Page 54: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Wet DNA fibers

X-ray beam

The pattern represents theatomic array in wet fibers.

X rays diffractedby DNA

(b) X-ray diffraction of wet DNA fibers

She worked in the same laboratory as Maurice Wilkins

She used X-ray diffraction to study wet fibers of DNA

Rosalind Franklin (early 1950s)

The diffraction pattern is interpreted (using mathematical theory)

This can ultimately provide information concerning the

structure of the molecule

Figure 9.12b

54

Page 55: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

She made marked advances in X-ray diffraction techniques with DNA

The diffraction pattern she obtained suggested several structural features of DNA Helical More than one strand 10 base pairs per complete turn

55

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56

Page 57: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Chargaff pioneered many of the biochemical techniques for the isolation, purification and measurement of nucleic acids from living cells

Erwin Chargaff (1950)

57

It was known that DNA contained the four bases: A, G, C and T

Page 58: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

• Experimental tools:– E. coli and many other organisms– DNA extraction protocol– Protease– Perchloric acid– Paper chromatography– Spectroscopy

58

Page 59: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

The Hypothesis

An analysis of the base composition of DNA in different species may reveal important features about the structure of DNA– Chargaff analyzed the base composition of

DNA isolated from many different species

Testing the Hypothesis

Refer to Figure 9.13

59

Page 60: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.13

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Experimental level Conceptual level

For each type of cell, extract the chromosomal material. This can bedone in a variety of ways, including theuse of high salt, detergent, or mild alkalitreatment. Note: The chromosomescontain both DNA and protein.

1.

Remove the protein. This can be done in several ways, including treament withprotease.

2.

Hydrolyze the DNA to release the bases from the DNA strands. A common wayto do this is by strong acid treatment.

3.

Separate the bases by chromatography. Paper chromatography provides an easyway to separate the four types of bases.(The technique of chromatography isdescribed in the Appendix.)

4.

Extract bands from paper into solutions and determine the amounts of each baseby spectroscopy. Each base will absorblight at a particular wavelength. Byexamining the absorption profile of asample of base, it is then possible tocalculate the amount of the base.(Spectroscopy is described in theAppendix.)

5.

Compare the base content in the DNA from different organisms.

6.

Solution of chromosomalextract

DNA

DNA + proteins

Individual bases

Origin

Protease

AcidA

A

AAA A

AA

CC

C CC CC

GG GG G GG

TT TT T

T

T

T

G

G

G

C

C

C

y

60

Page 61: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

The Data

61

Page 62: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Interpreting the Data

The data shown in Figure 9.13 are only a small sampling of Chargaff’s results

The compelling observation was that Percent of adenine = percent of thymine Percent of cytosine = percent of guanine

This observation became known as Chargaff’s rule It was a crucial piece of evidence that Watson and Crick

used to elucidate the structure of DNA

62

Page 63: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Familiar with all of these key observations, Watson and Crick set out to solve the structure of DNA They tried to build ball-and-stick models that

incorporated all known experimental observations

A critical question was how the two (or more strands) would interact An early hypothesis proposed that DNA strands interact

through phosphate-Mg++ cross-links Refer to Figure 9.14

Watson and Crick (1953)

63

Page 64: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.14

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Sugar

–O

O O

OP

Base

Sugar

Base

Sugar

O

OOP

Base

Sugar

BaseO–

Mg2+

64

WRONG! (But why is it appealing?)

Page 65: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

65

Watson and Crick weren’t the only ones making incorrect guesses for the structure of DNA!

Triple helix

Page 66: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Watson and Crick went back to the ball-and-stick units

They then built models with the Sugar-phosphate backbone on the outside Bases projecting toward each other

They first considered a structure in which bases form H bonds with identical bases in the opposite strand ie., A to A, T to T, C to C, and G to G

Model building revealed that this also was incorrect

66

Page 67: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

They then realized that the hydrogen bonding of A to T was structurally similar to that of C to G So they built ball-and-stick models with AT and CG

interactions between the two DNA strands These were consistent with all known data about DNA

structure Refer to Figure 9.15

Published model in April 1953 (Nature) Watson, Crick and Maurice Wilkins were

awarded the Nobel Prize in 1962 Rosalind Franklin died in 1958, and Nobel prizes

are not awarded posthumously

67

Page 68: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

(b) Original model of the DNA double helix

(a) Watson and Crick

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

© Barrington Brown/Photo Researchers

© Hulton|Archive by Getty Images

Figure 9.15 68

Page 69: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

69

Practice Problem(One correct answer):

A species of frog has a genome that contains 12% G. What percent T does it contain?a. 12%b. 38%c. 44%d. 76%e. 88%

Page 70: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

General structural features (Figures 9.16 & 9.17)

The DNA Double Helix

Two strands are twisted together around a common axis

There are 10 bases and 3.4 nm per complete turn of the helix

The two strands are antiparallel One runs in the 5’ to 3’ direction and the other 3’ to 5’

The helix is right-handed As it spirals away from you, the helix turns in a

clockwise direction70

Page 71: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

General structural features (Figures 9.16 & 9.17)

The double-bonded structure is stabilized by 1. Hydrogen bonding between complementary

bases A bonded to T by two hydrogen bonds C bonded to G by three hydrogen bonds

2. Base stacking Within the DNA, the bases are oriented so that the

flattened regions are facing each other

71

Page 72: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.16

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

H

N HN

N

N

G

NH2

H

P

S

P

SP

5¢end

3¢end

H NH2

N

N

HH

H

HH

OOO

O

P CH2

O

HH

H

OH

HH

OOO

O

P CH2

O

HH

H

HH

OOO

O

P CH2

O

CH2

HH

H

HH

OOO

O

P

O

T

Key Features

• Two strands of DNA form a right-handed double helix.

• The bases in opposite strands hydrogen bond according to theAT/GC rule.

• The 2 strands are antiparallel with regard to their 5′ to 3′ directionality.

• There are ~10.0 nucleotides in each strand per complete 360° turn ofthe helix.

2 nm

One nucleotide0.34 nm

One complete turn 3.4 nm

TA

G C

T AP

P

P

P

P

S

S

S

S

S

S

S

S

S

A

C

G

C G

C G

G C

G C

GC

G C

C

P

PS

P

P

P

P

P

S

S

S

S

S

PP

P

P

P

P

P

P

P

P

S

S S

S

S

S

S

S

S

P

S

S

P

P

P

S

S

S

S

P

G

S3¢5¢

S

A

P

P

C

T A

O

N

N

N

NA

H

H NH2

N

O

H

N

CH3

H

T

H

HH2N

N

N C

O

3¢end

5¢end

H

H

HH

OOO

O

PCH2

O

H

H

H

H

HH

OOO

O

PCH2

O

H

H

HH

OOO

O

PCH2

O

H

H

HH

OOO

O

PCH2

O

HO

N

O

H

N

CH3

H

T

O

NH N

N

N

G

H2N

H

H

H

N

N

N

N A

H

H2N H

-

-

-

-

-

-

-

-

-

-

72

Page 73: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

73

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Page 74: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

General structural features (Figures 9.16 & 9.17)

There are two asymmetrical grooves on the outside of the helix

1. Major groove 2. Minor groove

74

Page 75: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

(b) Space-filling model of DNA(a) Ball-and-stick model of DNA

Minorgroove

Majorgroove

Minorgroove

Majorgroove

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

© Laguna Design/Photo Researchers

Figure 9.17 75

Page 76: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Proteins can bind within these grooves They can thus interact with a particular sequence of bases to

“read” the DNA sequence

76

E.g. Restriction enzymes like Eco RI

Palindrome

Primitive immune system

4 + 2 Rule

Page 77: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

77

“Homodimerizes” then binds in successive major grooves to cut each DNA strand

Page 78: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

78

To calculate average distance between cut sites:

4(# bp in recognition site)

For Eco RI = 46 = 4,096 bp/cut

To calculate number of cuts/genome:

(bp in genome)/4(# bp in recognition site)

Eco RI cuts the human genome:

(3.2 x 109 bp)/4096 bp/cut = 780,000 cuts

Page 79: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

To fit within a living cell, the DNA double helix must be extensively compacted into a 3-D conformation This is aided by DNA-binding proteins

Refer to 9.20

The Three-Dimensional Structure of DNA

79

Page 80: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Figure 9.20

DNA wound around histone

proteins

Radial loops(300 nm in diameter)

Metaphasechromosome

DNA(2 nm in diameter)

Nucleosomes(11 nm in diameter)

Histoneprotein

Each chromatid(700 nm in diameter)

30 nm fiber

80

Page 81: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

81

1 meter DNA/cell1013 cells/human

1013 meters of DNA/human3.8 x 108 meters from Earth to Moon

If uncoiled, DNA from a single human would stretch from earth to moon and back ~13,000 times!!

Page 82: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

The primary structure of an RNA strand is much like that of a DNA strand Refer to Figure 9.21 vs. 9.10

RNA strands are typically several hundred to several thousand nucleotides in length

In RNA synthesis, only one of the two strands of DNA is used as a template

RNA Structure

82

Page 83: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Figure 9.21

Adenine (A)

Guanine(G)

Uracil (U)

BasesBackbone

Cytosine (C)

O

HHHH

OOO

O–

P CH2

O–

HHHH

OOO

O

P CH2

O–

NH2

H

N

HHHH

OOO

O

P CH2

O–

H

H

HH

OH

HH

OOO

O

P CH2

O–

Sugar (ribose)

Phosphate

5′

4′ 1′

2′3′

5′

4′ 1′

2′3′

5′

4′ 1′

2′3′

5′

4′ 1′

2′3′OH

OH

OH

OH

RNAnucleotide

Phosphodiesterlinkage

3′

5′

NH2

OH

H

H

O

NH O

N

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

NN

N

N

N

N

N N

N

NH2

H

H

83

Page 84: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Although usually single-stranded, RNA molecules can form short double-stranded regions This secondary structure is due to complementary base-

pairing A to U and C to G

This allows short regions to form a double helix

RNA double helices typically Are right-handed Have the A form with 11 to 12 base pairs per turn

Different types of RNA secondary structures are possible Refer to Figure 9.22

84

Page 85: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

A U

A U

U A

G C

C G

C G

A U

U A

U A

C G

C G

C G

C G

C G

A

A

U

U

G

G

C

C

C

(a) Bulge loop (b) Internal loop (c) Multibranched junction (d) Stem-loop

G

C

C

G

U

AA

U

G

C

G

C

C

GA

UA U

A U

G C

A U

U A

G C

C G

C G

G

C

G

C

C

GA

U

A

U

G

C

C

G

C

GA

U

A

U

A

U

U

G

G

C

C

CA U

A U

G C

C G

A

AAU

U

U

U

G

G

C

C

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.Figure 9.22

Also called hair-pin

Complementary regions

Non-complementary regions

Held together by hydrogen bonds

Have bases projecting away from double stranded regions

85

Page 86: LECTURE 1 Introduction to DNA and RNA (Chapter 09) 1

Many factors contribute to the tertiary structure of RNA For example

Base-pairing and base stacking within the RNA itself

Interactions with ions, small molecules and large proteins

Figure 9.23 depicts the tertiary structure of tRNAphe

The transfer RNA that carries phenylalanine

Molecule contains single- and double-stranded regions

These spontaneously fold and interact to

produce this 3-D structure

Figure 9.23(a) Ribbon model

3′ end(acceptorsite)5′ end

Double helix

Double helix

Anticodon

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

86