Chapter 10

Molecular Biology of the Gene

PowerPoint Lectures for Biology: Concepts and Connections, Fifth Edition Campbell, Reece, Taylor, and Simon

Lectures by Chris Romero

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

10.2 DNA and RNA are polymers of nucleotides

Nucleic acids are polynucleotides made of long chains of nucleotide monomers Nitrogenous bases
Single-ring pyrimidines: thymine (T), cytosine ( C) Double-ring purines: adenine (A), guanine (G)

Sugar-phosphate backbone
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 DNA and RNA are identical except for two things Nitrogenous bases
DNA: A, C, G, T RNA: A, G, C, U

Sugars
DNA: deoxyribose

RNA: ribose
Animation: DNA and RNA Structure
Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-2a

Sugar-phosphate backbone

Phosphate group
A Nitrogenous base Sugar A

C DNA nucleotide

Nitrogenous base (A, G, C, or T) Phosphate group

T
Thymine (T)

G Sugar (deoxyribose)

T DNA nucleotide DNA polynucleotide

LE 10-2b

Thymine (T)

Cytosine (C)

Adenine (A) Purines

Guanine (G)

Pyrimidines

LE 10-2c

Nitrogenous base (A, G, C, or U) Phosphate group

Uracil (U)

Sugar (ribose)

LE 10-2d
Key
Hydrogen atom Carbon atom Nitrogen atom Oxygen atom

Phosphorus atom

10.3 DNA is a double-stranded helix

James Watson and Francis Crick worked out the three-dimensional structure of DNA, based on X-ray crystallography by Rosalind Franklin DNA consists of two polynucleotide strands wrapped around each other in a double helix Sugar-phosphate backbones are on the outside and nitrogenous bases on the inside

Animation: DNA Double Helix

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 Each base pairs with a complementary partner

A with T, and G with C

Hydrogen bonds between the bases hold the strands together

The Watson-Crick model of DNA suggested a molecular explanation for genetic inheritance

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LE 10-3c

Twist

LE 10-3d

G T

C A A T Hydrogen bond T G C G C A

Base pair C A C G

G T

T T

A A A T

G A Ribbon model

C T Partial chemical structure Computer model

DNA REPLICATION
10.4 DNA replication depends on specific base pairing
The Watson-Crick model of DNA structure suggested a mechanism for its replication DNA strands separate Enzymes use each strand as a template to assemble new nucleotides into complementary strands The mechanism of DNA replication is semiconservative Each new double helix consists of one old and one new strand
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LE 10-4a

A C G A T

T G C T A

A C G A T

T G C C

T G C T

A C G A T

T G C T A

A C G A T

T G C T A

Nucleotides Both parental strands serve as templates

Parental molecule of DNA

Two identical daughter molecules of DNA

 DNA replication is a complex process Some of the helical DNA molecule must untwist

Animation: DNA Replication Overview

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-4b

G C A T G C

C G A T

10.5 DNA replication: A closer look

DNA replication begins at specific sites (origins of replication) on the double helix Proteins attach and separate the strands Replication proceeds in both directions, creating replication bubbles
Parent strands open, daughter strands elongate

Replication occurs simultaneously at many sites

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LE 10-5a

Origin of replication

Parental strand Daughter strand

Bubble

Two daughter DNA molecules

 DNA's sugar-phosphate backbones are oriented in opposite directions The enzyme DNA polymerase adds nucleotides at only the 3 end
One daughter strand is synthesized as a continuous piece The other strand is synthesized as a series of short pieces

The two strands are connected by the enzyme DNA ligase

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-5b
5 end 3 end

P 4 3

5 1 A T 1

HO 2 3 5 4 P C G

2 P

P
G

P T

OH

3 end

5 end

LE 10-5c
DNA polymerase molecule
3 5

5 3

Parental DNA

Daughter strand synthesized continuously Daughter strand synthesized In pieces

5 3

DNA ligase
Overall direction of replication

Animation: Origins of Replication Animation: Leading Strand Animation: Lagging Strand Animation: DNA Replication Review

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

THE FLOW OF GENETIC INFORMATION FROM DNA TO RNA TO PROTEIN 10.6 The DNA genotype is expressed as proteins, which provide the molecular basis for phenotypic traits
The information constituting an organism's genotype is carried in its sequence of DNA bases A particular genea linear sequence of many nucleotidesspecifies a particular polypeptide

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The flow of genetic information 1. Transcription of the genetic information in DNA into RNA 2. Translation of RNA into the polypeptide

Beadle-Tatum one gene-one enzyme hypothesis

Studies of inherited metabolic disorders in mold suggested that phenotype is expressed through proteins
A gene dictates production of a specific enzyme The hypothesis has been restated to one geneone polypeptide

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

10.7 Genetic information written in codons is translated into amino acid sequences
Genetic information flows from DNA to RNA to protein Nucleotide monomers represent letters in an alphabet that can form words in a language Triplet code
Three-letter words (codons) Each word codes for one amino acid in a polypeptide
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LE 10-7a
DNA molecule

Gene 1

Gene 2

Gene 3

DNA strand A Transcription U RNA Codon Translation U U G G C C G U U U U A A C C G G C A A A A

Polypeptide Amino acid

10.8 The genetic code is the Rosetta stone of life

The genetic code specifies the correspondence between RNA codons and amino acids in proteins Includes start and stop codons

Redundant but not ambiguous

Nearly all organisms use exactly the same genetic code

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LE 10-8a
Second base

U
UUU Phe UUC UCC

C
UCU
UAU

A
UGU Tyr UAC Ser UAA Stop Stop UGC UGA UGG CGU His CGU

G U
Cys

C
Stop Trp

U
UUA Leu UUG CUU CUC UCG CCU CCC Leu CUA CUG AUU AUC lle CCA CCG ACU ACC UCA

A G U C

UAG CAU
CAC Pro CAA

Arg CGA Gln CAG AAU Asn AAC AGC AGA Lys Arg AGG GGU Asp

A G U
Ser

CGG
AGU

A
AUA AUG GUU
Met or start

Thr ACA ACG GCU GCC Val GUA GUG GCA GCG Ala GAA Glu GAG GGG GGA AAA AAG GAU GAC

A
G U C
Gly

GUC

GGC

A G

LE 10-8b
Strand to be transcribed

T DNA

Transcription

A RNA

Start codon

Stop codon

Translation

Polypeptide

Met

Lys

Phe

10.9 Transcription produces genetic messages in the form of RNA

One DNA strand serves as a template for the new RNA strand RNA polymerase constructs the RNA strand in a multistep process Initiation
RNA polymerase attaches to the promotor Synthesis starts
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 Elongation: RNA synthesis continues

RNA strand peels away from DNA template

DNA strands come back together in transcribed region Termination RNA polymerase reaches a terminator sequence at the end of the gene Polymerase detaches
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LE 10-9a

RNA nucleotides
RNA polymerase

Direction of transcription

Template strand of DNA

Newly made RNA

LE 10-9b
RNA polymerase DNA of gene Promoter DNA Initiation Terminator DNA

Elongation

Area shown In Figure 10.9A

Termination

Growing RNA

Completed RNA

RNA polymerase

Animation: Transcription

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

10.10 Eukaryotic RNA is processed before leaving the nucleus

The RNA that encodes an amino acid sequence is messenger RNA (mRNA) In prokaryotes, transcription and translation both occur in the cytoplasm In eukaryotes, RNA transcribed in the nucleus is processed before moving to the cytoplasm for translation

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 RNA Splicing Noncoding segments called introns are cut out Remaining exons are joined to form a continuous coding sequence

A cap and a tail are added to the ends

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LE 10-10
Exon Intron DNA Cap Transcription Addition of cap and tail Exon Intron Exon

RNA transcript

with cap and tail

Introns removed

Tail

Exons spliced together mRNA

Coding sequence Nucleus

Cytoplasm

10.11 Transfer RNA molecules serve as interpreters during translation

Transfer RNA (tRNA) molecules match the right amino acid to the correct codon tRNA is a twisted and folded single strand of RNA Anticodon loop at one end recognizes a particular mRNA codon by base pairing Amino acid attachment site is at the other end Each amino acid is joined to the correct tRNA by a specific enzyme

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-11a
Amino acid attachment site

Hydrogen bond

RNA polynucleotide chain

Anticodon

LE 10-11b
Amino acid attachment site

Anticodon

10.12 Ribosomes build polypeptides

A ribosome consists of two subunits

Each is made up of proteins and ribosomal RNA (rRNA)

The subunits of a ribosome Hold the tRNA and mRNA close together in binding sites during translation

Allow amino acids to be connected into a polypeptide chain

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-12a
tRNA molecules

Growing polypeptide

Large subunit

mRNA

Small subunit

LE 10-12b

tRNA-binding sites

Large subunit

mRNA binding site

Small subunit

LE 10-12c
Next amino acid to be added to polypeptide

Growing polypeptide

tRNA

mRNA

Codons

10.13 An initiation codon marks the start of an mRNA message

The initiation phase of translation Brings together mRNA, a specific tRNA, and the two subunits of a ribosome

Establishes exactly where translation will begin

Ensures that mRNA codes are translated in the correct sequence

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

 Initiation is a two-step process Step 1

mRNA binds to a small ribosomal subunit
Initiator tRNA, carrying the amino acid Met, binds to the start codon

Step 2
A large ribosomal subunit binds to the small one, forming a functional ribosome Initiator tRNA fits into one binding site; the other is vacant for the next tRNA
Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-13a

Start of genetic message

End

LE 10-13b

Initiator tRNA P site

U A C A U G U A C A UG

Large Ribosomal subunit A site

Start codon
mRNA Small ribosomal subunit

10.14 Elongation adds amino acids to the polypeptide chain until a stop codon terminates translation
Once initiation is complete, amino acids are added one by one in a three-step elongation process 1. Codon recognition
2. Peptide bond formation 3. Translocation Elongation continues until a stop codon reaches the ribosome's A site, terminating translation

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-14
Polypeptide
P site A site Anticodon mRNA Codons
Condon recognition

Amino acid

mRNA movement

Stop codon

Peptide bond formation New peptide bond

Translocation

Animation: Translation

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

10.15 Review: The flow of genetic information in the cell is DNA RNA protein
The sequence of codons in DNA, via the sequence of codons in RNA, spells out the primary structure of a polypeptide 1. Transcription of mRNA from a DNA template 2. Attachment of amino acid to tRNA 3. Initiation of polypeptide synthesis 4. Elongation 5. Termination
Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-15
DNA Transcription

mRNA

RNA polymerase Translation

mRNA is transcribed from a DNA template.

Amino acid

Enzyme

Each amino acid attaches to its proper tRNA with the help of a specific enzyme and ATP.

ATP tRNA Anticodon Initiator tRNA

U AC AU G

Start Codon mRNA

Large Initiation of ribosomal polypeptide synthesis subunit The mRNA, the first tRNA, and the ribosomal Sub units come together. Small ribosomal subunit New peptide bond forming

Growing polypeptide

Codons mRNA

Elongation A succession of tRNAs add their amino acids to the polypeptide chain as the mRNA is moved through the ribosome, one codon at a time.

Polypeptide

Termination
The ribosome recognizes a stop codon. The polypeptide is terminated and released.

Stop codon

10.16 Mutations can change the meaning of genes

Mutation: any change in the nucleotide sequence of DNA Caused by errors in DNA replication or recombination, or by mutagens Can involve large regions of a chromosome or a single base pair Can cause many genetic diseases, such as sickle-cell disease
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LE 10-16a

Normal hemoglobin DNA

Mutant hemoglobin DNA

mRNA G A A

mRNA G U A

Normal hemoglobin

Sickle-cell hemoglobin

Glu

Val

 Two general categories of genetic mutations Base substitutions replace one base with another
Most are harmful but may occasionally have no effect or be beneficial

Base insertions or deletions alter the reading frame

Result is most likely a nonfunctioning polypeptide

Mutagenesis caused by spontaneous error or a physical or chemical mutagen

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-16b
Normal gene

A mRNA Protein

Met

Lys

Phe

Gly

Ala

Base substitution

Met

Lys

Phe

Ser

Ala

Base deletion

U Missing

Met

Lys

Leu

Ala

His

MICROBIAL GENETICS 10.17 Viral DNA may become part of the host chromosome
Viruses are infectious particles consisting of nucleic acid enclosed in a protein capsid Viruses depend on their host cells for the replication, transcription, and translation of their nucleic acid DNA enters host bacterium, circularizes, and enters one of two pathways

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 Lytic cycle
Host produces more viruses

Host cell lyses (breaks open) to release new viruses

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 Lysogenic cycle
Phage DNA inserted by recombination into the host chromosome; is now a prophage Prophages replicated each time host cell divides; passed on to generations of daughter cells Does not destroy host Environmental signal may trigger switch from lysogenic to lytic cycle

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-17

Phage Attaches to cell Phage DNA

Bacterial chromosome

Cell lyses, releasing phages

Phage injects DNA Many cell divisions

Lytic cycle Phages assemble Phage DNA circularizes OR

Lysogenic cycle Prophage Lysogenic bacterium reproduces normally, replicating the prophage at each cell division

New phage DNA and proteins are synthesized

Phage DNA inserts into the bacterial chromosome by recombination

Animation: Simplified Viral Reproductive Cycle Animation: Phage T4 Lytic Cycle Animation: Phage Lambda Lysogenic and Lytic Cycles

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CONNECTION 10.18 Many viruses cause disease in animals

Structure of a virus that invades animal cells

Genetic material may be RNA (examples: flu, HIV) or DNA (examples: hepatitis, herpes) Protein coat
Sometimes a membranous envelope with glycoprotein spikes The envelope helps the virus enter and leave the host cell during its reproductive cycle
Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-18a
Membranous envelope

RNA

Protein coat

Glycoprotein spike

LE 10-18b
VIRUS Viral RNA (genome) Glycoprotein spike Protein coat Envelope Entry

Plasma membrane of host cell

Uncoating Viral RNA (genome) RNA synthesis by viral enzyme Protein synthesis mRNA RNA synthesis (other strand) Template New viral genome New viral proteins Assembly

Exit

CONNECTION 10.19 Plant viruses are serious agricultural pests

Most plant viruses

Have RNA genomes

Enter their hosts via wounds in the plant's outer layers May spread throughout the plant through plasmodesmata

There is no cure for most plant viruses

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LE 10-19

Protein

RNA

CONNECTION 10.20 Emerging viruses threaten human health

Emerging viruses have appeared suddenly or have recently come to the attention of scientists Examples: HIV, SARS, Ebola, West Nile

Processes contributing to emergence

Mutation

Contact between species

Spread from isolated populations
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10.21 The AIDS virus makes DNA on an RNA template

HIV, the AIDS virus, is a retrovirus Flow of genetic information is RNA _ DNA Inside a cell, HIV uses its RNA as a template for making DNA The enzyme reverse transcriptase catalyzes reverse transcription
Animation: HIV Reproductive Cycle
Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

LE 10-21a

Envelope Glycoprotein

Protein coat

RNA (two identical strands)

Reverse transcriptase

LE 10-21b

Viral RNA

CYTOPLASM NUCLEUS

DNA strand Doublestranded DNA Viral RNA and proteins

Chromosomal DNA Provirus DNA

RNA

10.22 Bacteria can transfer DNA in three ways

Bacteria can transfer genes from cell to cell by one of three processes Transformation: the uptake of foreign DNA from the surrounding environment

Transduction: transfer of bacterial genes by a phage

Conjugation: union of two bacterial cells and the transfer of DNA between them

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LE 10-22a
DNA enters cell

Fragment of DNA from another bacterial cell

Bacterial chromosome (DNA)

LE 10-22b
Phage

Fragment of DNA from another bacterial cell (former phage host)

LE 10-22c

Mating bridge

Sex pili

Donor cell (male)

Recipient cell (female)

 Once new DNA is in a bacterial cell, part of it may integrate into the recipient's chromosome Occurs by crossing over between the two molecules Leaves the recipient with a recombinant chromosome

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LE 10-22d

Donated DNA

Crossovers

Degraded DNA

Recipient cells chromosome

Recombinant chromosome

10.23 Bacterial plasmids can serve as carriers for gene transfer

The F factor is a piece of bacterial DNA Carries genes for things needed for conjugation

Contains an origin of replication

Can transfer chromosomal DNA by integrating into the donor bacterium's chromosome or entering the cell as a plasmid
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LE 10-23a
F factor (integrated)
Male (donor) cell Origin of F replication Bacterial chromosome F factor starts replication and transfer of chromosome

Recipient cell

Only part of the chromosome transfers

Recombination can occur

LE 10-23b
F factor (plasmid)

Male (donor) cell Bacterial chromosome

F factor starts replication and transfer

Plasmid completes transfer and circularizes

Cell now male

 Plasmids Small circular DNA molecules separate from the bacterial chromosome Can serve as carriers for the transfer of genes

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LE 10-23c

Plasmids

Colorized TEM 2,000