Chapter 12: Molecular Biology of

the Gene
BIOL 1010
Rasch

The Genetic Material

The middle of the twentieth century was an exciting period of

scientific
i ifi discovery.
di
On one hand,
hand geneticists were busy determining that DNA

(deoxyribonucleic acid) is the genetic material of living things.

On the other hand, biochemists were in a frantic race to describe

the structure of DNA.

The classic experiments performed during this era set the stage

p
in our knowledge
g of modern molecular biology.
gy.
for an explosion

The Genetic Material

When researchers began their work, they knew that the

genetic material must be

Able to store information that pertains to the development,

structure, and metabolic activities of the cell or organism;

structure
Stable so that it can be replicated with high accuracy during cell
division and be transmitted from generation to generation;
Able to undergo rare changes called mutations that provide the
genetic variability required for evolution to occur.

Transformation of Bacteria
During the late 1920s, the bacteriologist Frederick Griffith

was attempting to develop a vaccine against Streptococcus

pneumoniae which causes pneumonia in mammals.
In 1931, he performed a classic experiment with the

bacterium..

Transformation of Bacteria
He noticed that when these bacteria are grown on culture

plates, some, called S strain bacteria, produce shiny, smooth

colonies, and others, called R strain bacteria, produce
colonies that have a rough appearance.
appearance
Under the microscope,
p , S strain bacteria have a capsule
p

(mucous coat) that makes them smooth but R strain bacteria

do not.

Transformation of Bacteria
When Griffith injected mice with the S strain of bacteria, the

mice died, and when he injected mice with the R strain, the
mice did not die.
In an effort to determine whether the capsule alone was

responsible
p
for the virulence (ability
(
y to kill)) of the S strain
bacteria, he injected mice with heat-killed S strain bacteria.
The mice did not die.

Transformation of Bacteria
Finally, Griffith injected the mice with a mixture of heat-

killed S strain and live R strain bacteria.

Most
M unexpectedly,
dl the
h mice ddiedand
d
d living
l
S strain

bacteria were recovered from the bodies!

Griffith concluded that some substance necessary for the

bacteria to pproduce a capsule

p
and be virulent must have
passed from the dead S strain bacteria to the living R strain
bacteria so that the R strain bacteria were transformed

Transformation of Bacteria
This change in the phenotype of the R strain bacteria must be

due to a change in their genotype.

Indeed,
I d d couldn't
ld ' the
h transforming
f
substance
b
that
h passedd ffrom

S strain to R strain be genetic material?

Reasoning such as this prompted investigators at the time to

begin
g lookingg for the transformingg substance to determine
the chemical nature of the genetic material.

Transformation of Bacteria

Avery, McLeod, and McCartys

E
Experiment
i
t
In the 1940s it was known that the genes are on the

chromosomes and that the chromosomes contain both

proteins and nucleic acids.
Investigators were having a much heated debate about

whether pprotein or DNA was the ggenetic material..

Avery et al.
al
Many thought that the protein component of chromosomes

must be the genetic material because proteins contain up to

20 different amino acids that can be sequenced in any
particular way.
way
On the other hand,, nucleic acidsDNA and RNAcontain

only four types of nucleotides as basic building blocks.

Some argued that DNA did not have enough variability to be

able to store information and be the genetic material.

Avery et al.
al
In 1944, Oswald Avery, Colin MacLeod and Maclyn

McCarty, published a paper demonstrating that the

transforming substance that allows Streptococcus to produce
a capsule and be virulent is DNA.
DNA
This meant that DNA is the ggenetic material..

Avery et al.
al
Here is what they found out:
DNA from S strain bacteria causes R strain bacteria to be

transformed
f
d so that
h they
h can produce
d a capsule
l andd bbe
virulent.
The addition of DNase, an enzyme that digests DNA,

p
prevents
transformation from occurring.
g This supports
pp
the
hypothesis that DNA is the genetic material.

Avery et al.
al
The molecular weight of the transforming substance is large.

This suggests the possibility of genetic variability.

The
Th addition
dd
off enzymes that
h degrade
d
d proteins has
h no effect
ff

on the transforming substance nor does RNase, an enzyme

that digests
g
RNA.. This shows that neither protein
p
nor RNA is
the genetic material.

Avery et al.
al
These experiments showed that DNA is the transforming

substance and, therefore, the genetic material.

Although
Al h h some scientists remainedd skeptical,
k
l many ffelt
l that
h

the evidence for DNA being the genetic material was

overwhelming.
g.

Hershey and Chase

Hershey and Chase used a virus called a T phage, composed

off radioactively
di ti l llabeled
b l d DNA andd capsid
id coatt proteins,
t i tto
infect E. coli bacteria.

They discovered that the radioactive tracers for DNA, but

not protein, ended up inside the bacterial cells, causing them

t bbecome transformed.
to
t f
d

Since only the genetic material could have caused this

transformation, Hershey and Chase determined that DNA

must be the genetic material.

Green Fluorescent Protein

Because the code for living things is based on the same four

nucleic acid bases of A, G, C, and T, and genes are made from

this code, it is conceivable to take genes from one organism
and put them into another.
another
Transformation of organisms,
g
, resultingg in ggeneticallyy

modified organisms (GMOs), is an invaluable tool in modern

biotechnology today.

Green Fluorescent Protein

Early biotechnologists seeking a dramatic way to show the

possibility of gene transfer between different organisms took

a jellyfish gene that codes for a green fluorescent protein
(GFP) and started transforming different organisms with it.
it
When this ggene is transferred to another organism,
g
, the

organism glows in the dark!

Structure of DNA
DNA contains:
Two nucleotides with purine bases
Adenine (A)
Guanine (G)
( )
Two nucleotides with pyrimidine bases
Thymine (T)
Cytosine
C
(C)

Erwin Chargaff used new chemical techniques developed in

the 1940s to analyze in detail the base content of DNA.

Structure of DNA
Some speciesE. coli and Zea mays (corn), for example

do have approximately 25% of each type of nucleotide, most

do not.
The % of each type of nucleotide differs from species to

species.
p
.
The nucleotide content of DNA is not fixed across species,
p
,

and DNA does have the variability between species required

for it to be the genetic material.

Structure of DNA
The percentage of A always equals the percentage of T, and

the percentage of G equals the percentage of C.

If the
h percentage off A + T equals
l 40%
40%, then
h the
h percentage

of G + C would equal 60%.

These relationships are called Chargaff's rules.

Chargaffs Rules
The amount of A, T, G, and C in DNA varies from species to

species.
In
I eachh species, the
h amount off A = T andd the
h amount off G =

C.

X Ray Diffraction of DNA

X-Ray
Rosalind Franklin studied the structure of DNA using X-rays.
She found that if a concentrated, viscous solution of DNA is

made, it can be separated into fibers.

Under the right conditions, the fibers can produce an X
X-ray
ray
diffraction pattern.
She produced X-ray diffraction photographs.
This
Thi provided
id d evidence
id
that
h DNA hhadd the
h ffollowing
ll i ffeatures:

DNA is a helix.
Some portion of the helix is repeated.
A colleague of Franklins, Wilkins, showed her photo to James Watson,
who understood its significance about DNAs structure.

Watson and Crick

Double helix model is similar to a twisted ladder.
Sugar-phosphate backbones make up the sides.
Hydrogen-bonded bases make up the rungs.
The two DNA strands are antiparallel.
antiparallel
Information stored in DNA must be read in the 5 to 3

direction so DNA is replicated in a 5 to 3 direction.

Complementary base pairing ensures that a purine is always

bonded to a pyrimidine (A with T, G with C).

They received a Nobel Prize in 1962

Watson and Crick

This model also agreed with Chargaff's rules, which states

that A = T and G = C.
This
Th so-called
ll d complementary
l
bbase pairing means that
h a

purine is always bonded to a pyrimidine.

This antiparallel pairing arrangement of the two strands

ensures that the bases are oriented pproperly

p y so that theyy can
interact.

Replication of DNA
DNA replication is the process of copying a DNA molecule.
Semiconservative replication: Each strand of the original

ddouble
bl helix
h l ((parentall molecule)
l l ) serves as a template
l ((mold
ld
or model) for a new strand in a daughter molecule.

DNA Replication
Replication requires the following steps:

Unwinding:The old strands that make up the parental DNA

molecule are unwound and unzipped (i.e., the weak

hydrogen bonds between the paired bases are broken).
broken)
A special enzyme called helicase unwinds the molecule.
molecule

DNA Replication
Complementary base pairing: New free nucleotides, always

present in the nucleus, are paired with nucleotides on the

parental strands, A with T, and G with C.
Joining:The complementary nucleotides paired with the

p
parental
strands are connected to each other to form a
connected chain. Each daughter DNA molecule now contains
an old strand and a newly synthesized strand.

DNA Replication
Steps 2 and 3 are carried out by an enzyme complex

called DNA polymerase.

DNA polymerase
l
works
k in the
h test tube
b as wellll as in cells.
ll

Prokaryotic DNA Replication

Bacteria have a single circular loop of DNA.
Replication moves around the circular DNA molecule in

both directions.
It
I produces
d
two identical
d
l circles.
l
The process begins at the origin of replication.
Replication
R li ti ttakes
k about
b t 40 minutes,
i t bbutt the
th cellll divides
di id every
20 minutes.
A new round of replication
p
can begin
g before the pprevious round

is completed.

Prokaryotic DNA Replication

The process begins at the origin of replication, a specific site

on the bacterial chromosome.

The
Th strands
d are separatedd andd unwound,
d andd a DNA

polymerase enzyme binds to each side of the opening and

begins
g the copying
py g process.
p
.
When the two DNA p
polymerases
y
meet at a termination

region, replication is halted, and the two copies of the

chromosome are separated.

Eukaryotic DNA Replication

DNA replication begins at numerous points along each linear

chromosome.
DNA unwinds and unzips into two strands.
Each
E h old
ld strandd off DNA serves as a template
l ffor a new
strand.
Complementary base pairing forms a new strand paired with
each old strand.
Requires enzyme
y DNA polymerase
y

There is a V shape wherever DNA is being replicated. This is

called a replication fork.

Eukaryotic DNA Replication

Replication bubbles spread bidirectionally, until they meet.
The complementary nucleotides are joined to form new

strands.
Replication is semiconservative:

One original strand is conserved in each daughter molecule:

each daughter double helix has one parental strand and one
newly synthesized strand.
strand

Polymerase is unable to replicate the ends of chromosomes

(telomeres).

Telomeres are copied by the enzyme telomerase.

Unregulated telomerase activity negatively affects cell function.

Eukaryotic DNA Replication

Therefore, eukaryotic cells complete the replication of the

diploid amount of DNA (in humans, over 6 billion base pairs)

in a matter of hours!

Accuracy of Replication
Accuracy of Replication
DNA polymerase is very accurate, yet makes a mistake about

once per 100,000 base pairs.

Capable of identifying and correcting errors

How DNA Replication Works

During replication, DNA polymerase needs a place to start

joining new nucleotides together.

It recognizes the
h OH
O chemical
h
l group at the
h 3 endd off an

existing nucleic acid chain, which can be DNA or RNA, and

it begins
g synthesizing
y
g from there..
Duringg DNA replication,
p
, an RNA-producing
p
g enzyme
y makes

a short primer that has the necessary 3 OH group on the

end.

How DNA Replication Works

DNA polymerase recognizes that target and begins DNA

synthesis, allowing new nucleotides to form complementary

base pairs with the old strand and connecting the new
nucleotides together in a chain.
chain

How DNA Replication Works

As a helicase enzyme unwinds DNA, it creates two

replication forks that move away from each other.

Each
E h off the
h parentall strands
d in a fforkk is accessible
bl ffor

complementary base pairing with new nucleotides and

therefore synthesis
y
of a new strand..
Bindingg proteins
p
coat the newlyy formed,, single-stranded
g

regions and prevent them from reattaching to each other.

How DNA Replication Works

The parental strands are antiparallel to each other, and each

of the new daughter strands must also be antiparallel to their

matching parental strandwhich creates a problem.
The new strand that gets made in the same direction as the

fork is movingg is called the leadingg strand..

How DNA Replication Works

The other new strand in the fork must be synthesized in a

direction opposite fork movement, which requires DNA

polymerase to periodically start and stop.
This strand is called the lagging strand.
Replication of this lagging strand is therefore made in

segments
g
called Okazaki fragments.
g

How DNA Replication Works

Replication is complete only when the RNA primers are

removed.
d

This works out well for the lagging strand.

strand
While checkingg to make sure bases are properly
p p y matched upp

(proofreading), DNA polymerase removes the RNA primers

and replaces them with the proper DNA nucleotides.

Another enzyme, called DNA ligase, joins the fragments,

creating a seamless DNA molecule.

RNA
RNA (ribonucleic acid) is a polymer composed of

nucleotides.
l tid

The nucleotides in RNA,

RNA however,
however contain the sugar ribose

and the bases adenine (A), cytosine (C), guanine (G),

and uracil (U).

In RNA, the base uracil replaces the thymine found in DNA.

RNA is single stranded and does not form a double helix in

the same manner as DNA

RNA
There are three major classes of RNA.
Messenger RNA (mRNA) takes a message from DNA in the

nucleus
l to the
h ribosomes
b
in the
h cytoplasm.
l
Transfer RNA (tRNA) transfers amino acids to the
ribosomes.
ribosomes
Ribosomal RNA (rRNA), along with ribosomal proteins,
makes upp the ribosomes,, where ppolypeptides
yp p
are synthesized.
y

The Genetic Code

During translation the mRNA transcript is read by a

ribosome and converted into the sequence of amino acids in a

polypeptide.
Together, the flow of information from DNA to RNA to

p
protein
to trait is known as the central dogma
g of molecular
biology.

The Genetic Code

The unit of the genetic code consists of codons, each of which is a

unique arrangement of symbols.

symbols
Each of the 20 amino acids found in proteins is uniquely specified
by one or more codons.

The symbols used by the genetic code are the mRNA bases.
Function as letters of the genetic alphabet
Genetic alphabet has only four letters (U, A, C, G)
Codons in the genetic code are all three bases (symbols) long.
They function as words of genetic information.
Permutations:
There are 64 possible arrangements of four symbols taken three at a time.
They are often referred to as triplets.
Genetic language only has 64 words.

Finding the Genetic Code

In 1961, Marshall Nirenberg and J. Heinrich Matthei

performed an experiment that laid the groundwork for

cracking the genetic code.
First, they found that a cellular enzyme could be used to

construct a synthetic
y
RNA (one
(
that does not occur in cells),
),
and then they found that the synthetic RNA polymer could
be translated in a test tube that contains the cytoplasmic
contents off a cell.
ll

Finding the Genetic Code

Their first synthetic RNA was composed only of uracil, and

the protein that resulted was composed only of the amino

acid phenylalanine.
Therefore, the mRNA codon for phenylalanine was known to

be UUU..
Later,, theyy were able to translate just
j three nucleotides at a

time; in that way, it was possible to assign an amino acid to

each of the mRNA codons

Features of the Genetic Code

Universal

With few exceptions,

exceptions all organisms use the code the same way
way.
They encode the same 20 amino acids with the same 64 triplets.

Degenerate (redundant)

There are 64 codons available for 20 amino acids.

acids
Most amino acids are encoded by two or more codons.

Unambiguous (codons are exclusive)

None
N
off th
the codons
d code
d ffor two
t or more amino
i acids.
id
Each codon specifies only one of the 20 amino acids.

Contains start and stop signals

Punctuation
P
i codons
d
Like the capital letter we use to signify the beginning of a sentence,

and the period to signify the end

Transcription
A segment of DNA serves as a template for the production of

an RNA molecule.
The gene unzips and exposes unpaired bases.
I serves as a template
It
l ffor mRNA
RNA fformation.
Loose RNA nucleotides bind to exposed DNA bases using
the C=G and A=U rule.
rule
When the entire gene is transcribed into mRNA, the result is
a ppre-mRNA transcript
p of the gene.
g
The base sequence in the pre-mRNA is complementary to
the base sequence in DNA.

Transcription
A single chromosome consists of one very long molecule

encoding hundreds or thousands of genes.

The genetic information in a gene describes the amino acid
sequence of a protein.
protein
The information is in the base sequence of one side (the sense

strand) of the DNA molecule.

The gene is the functional equivalent of a sentence

Transcription
The segment of DNA corresponding to a gene is unzipped to

expose the bases of the sense strand.

The genetic information in the gene is transcribed (rewritten)

into an mRNA molecule.

molecule
The exposed bases in the DNA determine the sequence in
which the RNA bases will be connected together.
RNA polymerase connects the loose RNA nucleotides together.
The
h completed
l d transcript contains the
h information
f
from
f
the
h

gene, but in a mirror image, or complementary form.

Transcription
Pre-mRNA is modified (or processed) before leaving the

eukaryotic nucleus.
Modifications to the ends of the primary transcript:
Cap on the 5
5 end
The cap is a modified guanine (G) nucleotide.
It helps a ribosome determine where to attach when translation
begins.
begins
Poly-A tail of 150200 adenines on the 3 end
It facilitates the transport of mRNA out of the nucleus.
It inhibits
h b degradation
d
d
off mRNA by
b hydrolytic
hd l
enzymes

Transcription
Pre-mRNA is composed of exons and introns.
The exons will be expressed.
The introns, occur in between the exons.
The exons are expressed
expressed and the introns are in
in the way.
way
It allows a cell to pick and choose which exons go into a particular
mRNA.
RNA splicing:
li i
Primary transcript consists of:
Some segments that will not be expressed (introns)
Segments that will be expressed (exons)

Transcription
In higher eukaryotes removal is done by spliceosomes that

contain small nuclear RNAs (snRNAs).

A spliceosome uses a ribozyme (enzyme made of RNA rather

than just protein) to cut and remove the introns.

introns
Remaining exons are spliced back together.
The result is a mature mRNA transcript.

Transcription
In prokaryotes, introns are removed by self-splicingthat

is, the intron itself has the capability of enzymatically splicing

itself out of a pre-mRNA.

 DNA contains both exons (protein-

coding sequences) and introns (non

(nonprotein-coding sequences). Both of
these are transcribed and are present
in pre
pre-mRNA.
mRNA.

During processing, a cap and a poly-

A taill (a series off adenine

d
nucleotides) are added to the
molecule.

Introns get cut out and the exons get

spliced together by complexes called

spliceosomes. Once processing is
complete the mRNA molecule is
ready to leave the nucleus.
nucleus

Transcription
Functions of introns:
As organismal complexity increases
The number of protein-coding genes does not keep pace.
The p
proportion
p
of the ggenome that is introns increases.
Possible functions of introns
Exons might combine in various combinations.
Would
W ld allow
ll ddifferent
ff
mRNAs
RNA to result
l ffrom one segment off DNA
Introns might regulate gene expression.
Introns may encourage crossing-over during meiosis.
Exciting new picture of the genome is emerging.

Translation
Translation
The
Th sequence off codons
d iin th
the mRNA
RNA att a ribosome
ib
di
directs
t th
the

sequence of amino acids into a polypeptide.

A nucleic acid sequence is translated into a protein sequence.

Transfer RNA (or tRNA) molecules have two binding sites:

One associates with the mRNA transcript.
p
The other associates with a specific amino acid.
Each of the 20 amino acids in proteins associates with one or more of

64 types of tRNA.
The wobble hypothesis predicts that the third position in the tRNA
anticodon doesnt obey the A-U/G-C configuration rule and can be
variable..

Translation
An mRNA transcript associates with the rRNA of a ribosome

in the cytoplasm or a ribosome associated with the rough

endoplasmic reticulum.
The ribosome reads
reads the information in the transcript
transcript.
The ribosome directs various types of tRNA to bring in their
specific
p
amino acid fares.
.
The tRNA specified is determined by the code being
translated in the mRNA transcript.

Translation
tRNA molecules come in 64 different kinds.
All are very similar except that
One end bears a specific triplet (of the 64 possible) called the

anticodon
anticodon.
The other end binds with a specific amino acid type.
tRNA synthetases
y
attach the correct amino acid to the correct
tRNA molecule.
All tRNA molecules with a specific anticodon will always

b d withh the
bind
h same amino acid.
d

Role of tRNA
A tRNA that has the anticodon 5 AAG 3 binds to the mRNA

codon 5 CUU 3 and carries the amino acid leucine.

In
I the
h genetic code,
d 61 codons
d specify
f amino acids;
d the
h other
h

three serve as stop sequences

tRNA

Translation
Approximately 40 different tRNA molecules are found in

most cells.
There
Th are ffewer tRNAs
RNA than
h codons
d bbecause some tRNAs
RNA

can pair with more than one codon.

In 1966, Francis Crick observed this phenomenon and called

it the wobble hypothesis.

yp

Role of tRNA
He stated that the first two positions in a tRNA anticodon

pair obey the AU/GC configuration rule.

However,
H
the
h third
h d position can bbe variable.
bl
Some
S
tRNA molecules
l l can recognize
i as many as ffour

separate codons differing only in the third nucleotide.

Role of tRNA
The wobble effect helps ensure that despite changes in DNA

base sequences, the resulting sequence of amino acids will

produce a correct protein.
This is one of the reasons the genetic code is said to be

degenerate.
g
.

Role of tRNA
How does the correct amino acid become attached to the

correct tRNA molecule?

This
Th taskk is carriedd out bby amino acidcharging
d h
enzymes,

generically called aminoacyl-tRNA synthetases.

Just as a key fits a lock, each enzyme has a recognition site for

a pparticular amino acid to be jjoined to a specific

p
tRNA.

Role of tRNA
Leucine-tRNA synthetase attaches the leucine amino acid to

a tRNA with the correct anticodon.

This
Th is an energy-requiring process that
h uses ATP
ATP. A tRNA
RNA

with its amino acid attached is termed a charged tRNA.

Once the amino acidtRNA complex is formed, it is added

to the large
g ppool of charged
g tRNAs that exist in the
cytoplasm, where it can now be accessed by a ribosome
during protein synthesis.

Role of rRNA
Ribosomal RNA (rRNA):
Produced from a DNA template in the nucleolus of a nucleus.
Packaged with proteins into large and small ribosomal subunits.

A completed
l t d ribosome
ib
hhas th
three binding
bi di sites
it to
t facilitate
f ilit t

pairing between tRNA and mRNA.

The E ((for exit)) site
The P (for peptide) site, and
The A (for amino acid) site

Role of rRNA
Once translation begins, ribosomes can remain in the

cytoplasm, or they can become attached to endoplasmic

reticulum.

Role of rRNA
The large ribosomal subunit has enzyme activity that creates

a peptide bond between adjacent amino acids.

The peptide bond is created many times to produce a

polypeptide which folds into its three

three-dimensional
dimensional shape and
becomes a protein.

Translation
Initiation:
Components necessary for initiation are:
Small ribosomal subunit
mRNA transcript
p
Initiator tRNA, and
Large ribosomal subunit
Initiation factors (special proteins that bring the above together)
Initiator tRNA:
Always has the UAC anticodon
Always carries the amino acid methionine
Capable of binding to the P site

Initiation
A small ribosomal subunit attaches to mRNA transcript.
The beginning of transcript always has the START codon

(AUG).
An initiator tRNA (UAC) attaches to the P site.
site
A large ribosomal subunit joins the small subunit.

Elongation
Elongation refers to the growth in length of the

polypeptide.
RNA molecules bring their amino acid fares to the ribosome.
Rib
Ribosome reads
d a codon
d iin th
the mRNA.
RNA
Allows only one type of tRNA to bring its amino acid
Must have the anticodon complementary to the mRNA codon being read
Incoming tRNA joins the ribosome at its A site
Methionine of the initiator tRNA is connected to the amino acid

of the second tRNA by a peptide bond.

bond

Elongation
The second tRNA moves to P site (translocation).
The spent initiator tRNA moves to the E site and exits.
The ribosome reads the next codon in the mRNA.
It allows only one type of tRNA to bring its amino acid.
Must have the anticodon complementary to the mRNA codon being read
JJoins the ribosome at its A site
The dipeptide on the second amino acid is connected to the

amino acid of the third tRNA by a peptide bond.

Elongation

Met

peptide
Ser bond

tRNA

A tRNAamino acid
approaches the
ribosome and binds
at the A site.

Elongation

Asp

C A U
G U A

Two tRNAs can be at a

ribosome at one time;
the anticodons are
paired to the codons.

Val
Asp

Asp

C U G
G A C

Trp

peptide
bond

Val

C A U
G U A

G A C

Trp
Val

Val

Ala

Ala
Trp

C U G

C U G
G A C

G U A

Thr

Ser

Ser

Ala

anticodon

Trp

C A U
G U A

Met
Ser

C U G

Ala

Met

Met

asp

Peptide bond formation

attaches the peptide
chain to the newly
arrived amino acid.

G A C

A C C

The ribosome moves forward; the

empty
empty tRNA exits from the E site;
the next amino acidtRNA complex
is approaching the ribosome.

Termination
Termination:
Previous tRNA moves to the P site.
Spent tRNA moves to the E site and exits.
Ribosome reads the STOP codon at the end of the mRNA.
mRNA
UAA, UAG, or UGA
Does not code for an amino acid
A protein called a release factor binds to the stop codon and

cleaves the polypeptide from the last tRNA.

The ribosome releases the mRNA and dissociates into subunits.
subunits
The same mRNA may be read by another ribosome

Gene Expression
A gene has been expressed once its product, a protein (or an

RNA), is made and is operating in the cell.

For
F a protein, gene expression requires transcription andd

translation and it also requires that the protein be active.

Gene Expression
Translation occurs at ribosomes. Some ribosomes

(polyribosomes) remain free in the cytoplasm, and some

become attached to rough ER.
The first few amino acids of a polypeptide act as a signal

p p
peptide
that indicates where the ppolypeptide
yp p
belongs
g in the
cell or if it is to be secreted from the cell.

Gene Expression
Polypeptides that are to be secreted enter the lumen of the

ER by way of a channel, and are then folded and further

processed by the addition of sugars, phosphates, or lipids.
Transport vesicles carry the proteins between organelles and

to the pplasma membrane as appropriate

pp p
for that pprotein..

Structure of the Eukaryotic

Chromosome
Ch
Only in recent years have investigators been able to produce

models suggesting how chromosomes are organized.

A eukaryotic
k
chromosome
h
contains a single
l ddouble
bl hhelix
l

DNA molecule, but it is composed of more than 50%

p
protein.
.
Some of these p
proteins are concerned with DNA and RNA

synthesis, but a large majority, termed histones, play

primarily a structural role.

Structure of the Eukaryotic

Chromosome
Ch
The five primary types of histone molecules are designated

H1, H2A, H2B, H3, and H4.

Remarkably,
R
k bl the
h amino acidd sequences off H3 andd H4 vary

little between organisms.

Structure of the Eukaryotic

Chromosome
Ch
For example, the H4 of peas is only two amino acids different

from the H4 of cattle.

This
Th similarity
l
suggests that
h ffew mutations in the
h hhistone

proteins have occurred during the course of evolution and

that the histones,, therefore,, have essential functions for
survival.

Structure of the Eukaryotic

Chromosome
Ch
A human cell contains at least 2 m of DNA.
Yet, all of this DNA is packed into a nucleus that is about 5

m in ddiameter.
The
Th histones
hi t
are responsible
ibl ffor packaging
k i th
the DNA so th
thatt it

can fit into such a small space.

Structure of the Eukaryotic

Chromosome
Ch
First, the DNA double helix is wound at intervals around a

core of eight histone molecules (two copies each of H2A,

H2B, H3, and H4), giving the appearance of a string of beads.
Each bead is called a nucleosome, and the nucleosomes are

said to be jjoined byy linker DNA..

Structure of the Eukaryotic

Chromosome
Ch
This string is compacted by folding into a zigzag structure,

further shortening the DNA strand.

Histone
H
H1 appears to mediate
d
this
h coiling
l process.
The
Th fib
fiber th
then lloops back
b k andd forth
f th iinto
t radial
di l lloop.

Structure of the Eukaryotic

Chromosome
Ch
This loosely coiled euchromatin represents the active

chromatin containing genes that are being transcribed.

The
Th DNA off euchromatin
h
may bbe accessedd bby RNA

polymerase and other factors that are needed to promote

transcription.
p .
Recent research seems to indicate that regulating
g
g the level of

compaction of the DNA is an important method of

controlling gene expression in the cell.

Structure of the Eukaryotic

Chromosome
Ch
Under a microscope, one often observes dark-stained fibers

within the nucleus of the cell.

These
Th areas within
h the
h nucleus
l represent a more highly
h hl

compacted form of the chromosome called heterochromatin.

Structure of the Eukaryotic

Chromosome
Ch
Most chromosomes exhibit both levels of compaction in a

living cell, depending on which portions of the chromosome

are being used more frequently.
Heterochromatin is considered inactive chromatin because

the ggenes contained on it are infrequently

q
y transcribed,, if at
all.

Structure of the Eukaryotic

Chromosome
Ch
Prior to cell division, a protein scaffold helps to further

condense the chromosome into a form that is characteristic

of metaphase chromosomes.
No doubt, compact chromosomes are easier to move about

than extended chromatin.. JJust liked rolled upp yyarn is easier

to move than yarn in a jumbled up string