CHAPTER 18 –

REGULATION
OF GENE
EXPRESSION
 Regulating PROKARYOTIC Gene Expression
 Both prokaryotes and
 Both prokaryotes and --Bacteria
Bacteriaoften
oftenrespond
respondto toenvironmental
environmentalchange
changeby by
eukaryotes
eukaryotesalter
altertheir
their regulating
regulatingat atthe
thelevel
levelofofTRANSCRIPTION!!
TRANSCRIPTION!!
patterns of gene --Natural
patterns of gene Naturalselection
selectionfavors
favorsbacteria
bacteriathat
thatexpress
expressonlyonlythose
those
expression
expressionin inresponse genes
response geneswhose
whoseproducts
productsare areneeded
neededby bythe
thecell.
cell.
to changes in --Metabolic
to changes in
environmental Metaboliccontrol
controloccurs
occursonontwo
twolevels.
levels.
environmental --First,
conditions. First,cells
cellscan
canadjust
adjustthe
theACTIVITY
ACTIVITYof ofenzymes
enzymes
conditions. already
 During development,
 During development,
alreadypresent.
present.This
Thismay
mayhappen
happenby byfeedback
feedback
inhibition,
inhibition,in inwhich
whichthetheactivity
activityof
ofthe
thefirst
firstenzyme
enzyme
gene
geneexpression
expressionmust must in
inaapathway
pathwayisisinhibited
inhibitedbybythe
thepathway’s
pathway’send end
bebecarefully
carefullyregulated
regulatedtoto
ensure product.
product.
ensurethat
thatthe
theright
right
genes are expressed --Second,
Second,cells
cellscan
canvary
varythe
theNUMBER
NUMBERof ofspecific
specific
genes are expressed
only ENZYME
ENZYMEMOLECULES
MOLECULESthey theymake
makeby byregulating
onlyatatthe
thecorrect
correcttime
time regulating
and in the correct place.
and in the correct place. gene
geneexpression.
expression.

The
Thebasic
basicmechanism
mechanismforforthe
thecontrol
controlof
ofgene
geneexpression
expressionininbacteria,
bacteria,known
knownas asthe
the
operon
operonmodel,
model,was
wasdescribed
describedby
byFrancois
FrancoisJacob
Jacoband
andJacques
JacquesMonod
Monodin in1961.
1961. Using
Using
these
theseoperons
operonsto
toalter
alterpatterns
patternsof
ofgene
geneexpression
expressionin
inprokaryotes
prokaryotesserves
servesan an
organism’s
organism’ssurvival
survivalbybyallowing
allowingananorganism
organismtotoadjust
adjustto
tochanges
changesin inthe
the
environmental
environmentalconditions.
conditions.
 Operons
Operons
Operonsare
arehow
how
prokaryotic TRP
TRPOperon
Operon==
prokaryoticgenes
genesare
are
controlled. makes
makes
controlled.
tryptophan
tryptophan
AAkeykeyadvantage
advantageofof
grouping LAC
LACOperon
Operon==
groupinggenes
geneswith
with
related breaks
breaksdown
relatedfunctions
functionsinto
into down
one lactose
onetranscription
transcriptionunit
unit lactose
isisthat
thataasingle
singleon-off
on-off Jacques Monod
switch
switchcancancontrol
controlaa
Francois Jacob Prokaryotic
cluster
clusterofoffunctionally
functionally Prokaryoticcells
cellscan
can
related
relatedgenes.
genes. control
controlmetabolism
metabolism
two
twoways:
ways:
1.1. Regulate
Regulateexpression
expression
of
ofgenes
genes(vary
(vary
number
numberof ofenzymes
enzymes
made)
made)
2.2. Adjust
Adjustthe
theactivity
activity
of
ofthe
theenzymes
enzymes
already
alreadypresent
present
(activators/
(activators/
inhibitors)
inhibitors)
 Parts of
an
Operon
They
Theyare
aremade
madeup upofof33parts:
parts:
1.1.Genes
Genesititcontrols
controls
(called
(calledstructural
structural
PROMOTER
PROMOTER== OPERATOR
OPERATOR==ON/ ON/OFF
OFF genes)
genes)
place
placewhere
wherethe
the Switch
Switch(located
(locatedwithin
within 2.2.Promoters
Promoters
RNA
RNApolymerase
polymerase the
thepromoter);
promoter);allows
allowsor
or 3.3.Operator
Operator(on/
(on/off
off
binds;
binds;includes
includesaa disallows
disallowsRNA
RNA switch)
switch)
TATA
TATAboxbox polymerase
polymerasetotobind
bind Recall:
Recall:Transcription
TranscriptionFactors
Factors
REPRESSOR
REPRESSOR==this thisbinds
bindsto
tothe
theoperator
operatorto
toblock
block bind
bindto
tothe
thepromoter
promoteroror
the
theattachment
attachmentof
ofRNA
RNAPolymerase
Polymerasewhen
whenititisisin
inthe
the TATA
TATAboxboxto
tohelp
helpRNA
RNA
active
activeform
form Polymerase
Polymerasebind
bind
The
Therepressor
repressorisiscoded
codedfor
forby
byaaregulatory
regulatorygene
genethat
thatisislocated
locatedaway
awayfrom
fromthe
the
operon.
operon. It has its own promoter. For the trp operon, the repressor is madein
It has its own promoter. For the trp operon, the repressor is made inthe
the
inactive
inactiveform,
form,and
andneeds
needstryptophan
tryptophanto tobecome
becomeactive.
active.SO,
SO,the
thegene
geneisisusually
usuallyON,
ON,
unless
unlessthe
therepressor
repressorgets
getsturned
turnedinto
intothe
theactive
activeform.
form.
 Trp Operon = Repressible

Repressible
Repressibleoperons
operonsare
arealways
always Bacteria
Bacteriasynthesize
synthesizetryptophan
tryptophanfrom
fromaa
ON
ONunless
unlessrepressed
repressed(switched
(switched pathway
pathwaythat
thatincludes
includes55enzymes.
enzymes. These
These
off)
off) enzymes
enzymesarearecoded
codedfor
forby
by55genes
genesall
allfound
found
together
togetherin
inthe
theTrp
TrpOperon.
Operon.
Therefore,
Therefore,the
theOPERON
OPERONisis
usually
usuallyOFF
OFF(unless
(unlessswitched
switchedon)
on) Feedback
FeedbackInhibition
Inhibition––ififmuch
muchtryptophan
tryptophanisis
present,
present,ititacts
actsas
asaaco-repressor.
co-repressor. ItItbinds
bindsto
to
All
Allthe
thegenes
genesare
arefound
foundtogether,
together, the
therepressor,
repressor,andandactivates
activatesit.
it. Then,
Then,the
the
so
sothat
thatONE
ONEoperator
operatorcontrols
controls repressor
repressorbinds
bindstotothe
theoperator
operatorandandblocks
blocks
expression
expressionof
ofALL
ALLof ofthe
thegenes
genes the
theattachment
attachmentof ofRNA
RNApolymerase
polymerase

Co-repressor
Co-repressor(ex.
(ex.
Tryptophan)
Tryptophan)turns
turns
genes
genesOFF
OFFby by
activating
activatingthe
the
repressor
repressor
IfIftryptophan
tryptophanisispresent,
present,
the
therepressor
repressorisis“active”,
“active”,
so
soititbinds
bindsto
tothe
thepromoter
promoter SO….
SO….
blocking
blockingthetheRNA No
RNA Notryptophan
tryptophan==repressor
repressor
polymerase. inactive
polymerase. inactive==operon
operonON
ON==making
making
tryptophan
tryptophan
Therefore,
Therefore,the
theproduction
production
of
oftryptophan
tryptophanisisstopped Lots
stopped Lotsof
oftryptophan
tryptophan==repressor
repressor
because
becausethere
thereisisalready activated
already activatedbybycorepressor
corepressor==operon
operon
enough
enoughininthe OFF
the OFF==nonotryptophan
tryptophanmade
made
environment.
environment.

SO…PRESENCE
SO…PRESENCEOF OF
TRYPTOPHAN
TRYPTOPHANTURNS
TURNS
THE
THEREPRESSOR
REPRESSORON,
ON,
WHICH
WHICHTURNS
TURNSTHE
THE
OPERON
OPERONOFFOFF

ENOUGH
ENOUGHTRYPTOPHAN
TRYPTOPHAN
IS
ISPRESENT
PRESENTSO
SOWE
WE
DON’T
DON’TNEED
NEEDTO
TOMAKE
MAKE
ANY
ANYMORE!
MORE!
Tryptophan
TryptophanOperon
Operon––
On
Onvs.
vs.Off
Off
 Lac Operon = Inducible
Inducible
InducibleOperons
Operonsare
arealways
alwaysOFF
OFFunless
unlessswitched
switchedON.
ON.

So
Sothe
therepressor
repressorisisnormally
normallyACTIVE,
ACTIVE,ititisisnormally
normallyrepressing
repressingthe
theoperon
operon(the
(the
regulatory
regulatorygene
genethat
thatencodes
encodesthe
therepressor
repressorencodes
encodesthe
theactive
activeconformation).
conformation). ItItisis
bound
boundtotothe
theoperator,
operator,and
andtherefore
thereforeblocks
blocksRNARNApolymerase.
polymerase.

The
TheLac
LacOperon
Operonbreaks
breaksdown
downlactose.
lactose. IfIfits
itsnot
notpresent
presentin
inthe
thebacteria's
bacteria'senvironment,
environment,
there
thereisisno
noneed
needto
tobreak
breakititdown.
down. Once
Onceititbecomes
becomesavailable,
available,the
theoperon
operonwould
wouldhave
havetoto
get
getswitched
switchedon onto
tobreak
breakititdown.
down.

The
Therepressor
repressorin inthe
theLac
Lac
Operon
Operonisismade
madein inthe
theactive
active
form,
form,so
soititisisnormally
normally
switched
switchedON.ON.
IfIflactose
lactoseisispresent,
present,ananisomer
isomerofofit,
it,allolactose,
allolactose, Remember,
Remember,these
theseoperons
operons
acts
actsasasan
aninducer.
inducer. ItItbinds
bindstotothe
therepressor,
repressor, code
codefor
forthe
themRNA
mRNAthat thatisis
which
whichinactivates
inactivatesit.
it. Now
Nowthat
thatthe
therepressor
repressornono going
goingto
togo
gototothe
the
longer
longerworks,
works,the
theoperon
operoncan canturn
turnon.on. Nothing
Nothing ribosomes
ribosomesto tomake
makethethe
isisbound
boundto tothe
theoperator,
operator,so soRNA
RNApolymerase
polymerase enzymes
enzymesthat
thatwill
willeither
either
can
canbind,
bind,and
andthe
thelactose
lactosecancanbebebroken
brokendown.
down. break
breakdown
downlactose,
lactose,oror
make
maketryptophan.
tryptophan.
An inducer inactivates the repressor
 Inducible vs. Repressible Operons

Inducible
InducibleOperons
Operons→ →
1.1. Repressors
Repressorsmade
madeininACTIVE
ACTIVEform form
2.2.Operon
Operonisisusually
usuallyOFF
OFF
3.3.When
Whenthe
therepressor
repressorisisinactivated
inactivatedby
byaamolecule,
molecule,then
thenthe
theoperon
operoncan
canbe
be
switched
switchedONON
4.4.Ex.
Ex.Lac
LacOperon
Operon→ →Lactose
Lactosemetabolism
metabolism

Repressible
RepressibleOperons
Operons→→
1.1.Repressors
Repressorsmade
madeininthe
theINACTIVE
INACTIVEformform
2.2.Operon
Operonisisusually
usuallyON
ON
3.3.When
Whenthe
therepressor
repressorisisswitched
switchedon,
on,ititbinds
bindsto
tothe
theoperator
operatorand
andblocks
blocks
RNA
RNAPolymerase
Polymerase, ,which
whichswitches
switchesthe
theoperon
operonOFFOFF
4.4.Ex.
Ex.Trp
TrpOperon
Operon→ →Synthesizing
Synthesizingtryptophan
tryptophan

Both
Bothof
ofthese
theseare
areexamples
examplesof
ofNEGATIVE
NEGATIVEcontrol
control––the
the
operon
operonis
isswitched
switchedOFF
OFFbybyan
anactive
activerepressor.
repressor.
 Positive Gene Regulation
Positive
PositiveControl
Control==something
somethingthat
thatbinds
bindsto
to cAMP
cAMP==cyclic
cyclicAMP;
AMP;accumulates
accumulates
the
theoperon
operondirectly
directlythat
thatswitches
switchesititON;
ON;the
the when
whenglucose
glucose(E(Esource)
source)drops
drops
degree
degreeof
oftranscription
transcriptiondepends
dependsononthethe CAP
CAP==Catabolite
CataboliteActivator
ActivatorProtein;
Protein;
concentration
concentrationofofother
othersubstances
substances activates
activatestranscription
transcriptioninitiation
initiationof
of
operons
operons

SO…Glucose
SO…Glucosedrops
drops==cAMP
cAMPincreases
increases==CAP
CAPbecomes
becomesactive
active==transcription
transcriptionisisON
ON
Lactose
Lactose(allolactose)
(allolactose)present
present Lactose
Lactosepresent
present→
→ON
ON Lactose
Lactosepresent
present→
→
→→Operon
Operonturned
turnedON
ON ON
ON
Glucose
Glucosepresent
present(LOW
(LOW
NO
NOLACTOSE
LACTOSE(allolactose)
(allolactose)→
→ cAMP)
cAMP)→ →CAP
CAPinactive
inactive, , No
NoGlucose
Glucose(HIGH
(HIGH
Operon
Operonturned
turnedOFF
OFF on
onaaLITTLE
LITTLE cAMP)
cAMP)→ →CAP
CAP
active,
active,on
onaaLOT
LOT
 Lac Operon – Dual Control
Negative Control → Repressor (presence/ absense
allolactose) = ON/ OFF SWITCH

Positive Control → CAP (level of transcription);

level of glucose and thus cAMP = VOLUME
CONTROL
 Regulating EUKARYOTIC Gene
 In
InPROKARYOTES,
PROKARYOTES,
Expression
they
theyregulate
regulategene
gene
expression at the
expression at the The differences
level
levelof
of between cell types
TRANSCRIPTION
TRANSCRIPTION are due to

 InInEUKARYOTES
EUKARYOTES differential gene
(greater
(greater expression, the
complexity),
complexity),they
they expression of
have
havethe
the different genes by
opportunity
opportunityto to cells with the
regulate
regulateatatmany
many same genome.
levels:
levels:
 Chromatin
Chromatin Problems with
Packing
Packing gene expression
 Transcription and control can
Transcription
 RNA Processing
RNA Processing lead to imbalance
 Translation
Translation
and disease,
 Post-translation including cancer.
Post-translation
 Structure of
Chromatin
DNA
DNAinineukaryotic
eukaryoticcells
cellsisis
packaged
packagedwith
withproteins
proteinsininaa
complex
complexcalled
calledchromatin.
chromatin.

Levels
LevelsofofChromatin
ChromatinPacking:
Packing:
1.1.Nucleosome
Nucleosome
2.2.30
30nm
nmchromatin
chromatinfibers
fibers
3.3.Looped
LoopedDomains
Domains
4.4.Chromosomes
Chromosomes

15
Nucleosomes and heterochromatin vs. euchromatin

Nucleosomes
Nucleosomesare arethe
thebasic
basicunit
unitof
ofDNA
DNA
packing;
packing; they
theyare
arecalled
called“beads
“beadsononaastring”
string”
because
becauseofofhow
howthey
theyappear;
appear;they
theyare
arecomposed
composed
of
ofhistones
histones(proteins)
(proteins)wrapped
wrappedin inDNA.
DNA.

Heterochromatin
Heterochromatin––veryverytightly
tightlycoiled;
coiled;
therefore
thereforeititisisNOT
NOTtranscribed
transcribed

Euchromatin
Euchromatin––“true
“truechromatin”;
chromatin”;ititisisless
less
compact
compactand
andtherefore
thereforethetheRNA
RNApolymerase
polymerase
can
canattach
attachand
andititcan
canget
gettranscribed
transcribed

16
 
 BOTH
BOTHof ofthese
theseprocesses
processesaffect
affectgene
geneexpression:
expression:

 Histone
HistoneAcetylation
Acetylation adding
addingacetyl
acetylgroups
groups(-(-
COCH
COCH3)to the histones (proteins); this INCREASES
Acetylation 3)to the histones (proteins); this INCREASES
TRANSCRIPTION
TRANSCRIPTIONbecause becauseititprovides
providesmore
morespace
space
for
forRNA
RNApolymerase
polymeraseto toattach
and 
 Histone
HistoneMethylation
Methylation
attach
adding
addingmethyl
methylgroups
groups(-(-
CH
CH3)to the histones; this DECREASES
Methylation 3)to the histones; this DECREASES
TRANSCRIPTION
TRANSCRIPTION

 DNA
DNAMethylation
Methylation adding
addingmethyl
methylgroups
groups(-CH
(-CH3)3)to
to
Acetylation
Acetylation==GOOD
GOOD==turns
turns DNA;
DNA;thisthisDECREASES
DECREASESTRANSCRIPTION;
TRANSCRIPTION;and andcancan
ON
ONtranscription
transcription SWITCH OFF (inactivate) genes  think Barr
SWITCH OFF (inactivate) genes  think Barr Bodies Bodies

Methylation
Methylation==BAD
BAD==turns
turns
OFF
OFFtranscription
transcription

So…chromatin
So…chromatincondensation
condensation
DECREASES
DECREASEStranscription,
transcription,
but
buthistone
histoneacetylation
acetylation
decreases
decreasesthe
theability
abilityof
of
chromatin
chromatintotocondense,
condense,so soitit
INCREASES
INCREASEStranscription
transcription
 Epigenetic Inheritance

 Inheritance
Inheritanceof oftraits
traitsby
by
mechanisms
mechanismsnot notdirectly
directly
involving
involvingthe thenucleotide
nucleotidesequence
sequence
isiscalled epigenetic inheritance.
called epigenetic inheritance.

 The
Theterm
termrefers
referstotochanges
changesto tothe
the
genome
genomethatthatdodoNOT
NOTinvolve
involveaa
change
changein inthe
thenucleotide
nucleotide
sequence.
sequence.

 Examples
Examplesof ofmechanisms
mechanismsthat that
produce
producesuchsuchchanges
changesare:
are:
 DNA methylation
DNA methylation
 Histone modification
Histone modification
 Inducers
Inducers
 Repressors
Repressors

 Epigenetic
Epigeneticvariations
variationsmaymayexplain
explain
why
whyoneoneidentical
identicaltwin
twinacquires
acquiresaa
genetically
geneticallybased
baseddisease,
disease,such
suchasas
schizophrenia, while another
schizophrenia, while another
does
doesnot,
not,despite
despitetheir
theiridentical
identical
genomes.
genomes.
 Control of gene
expression
In
Ineukaryotic
eukaryoticcells,
cells,gene
geneexpression
expressioncan
canbe
be
regulated
regulatedat atmany
manydifferent
differentpoints.
points.
--Initiation
InitiationofofTranscription
Transcription
--Post
Posttranscriptional
transcriptionalmodifications
modifications
(alternative
(alternativeRNA
RNASplicing)
Splicing)
--Initiation
InitiationofofTranslation
Translation
--Post-translational
Post-translational

19
 Control of
transcription
initiation

By
Byadding
addingadditional
additionaltranscription
transcriptionfactors;
factors;itit
can
canspeed
speedupupinitiation,
initiation,and
andthus
thusspeed
speedupup
transcription.
transcription.

Chromatin-modifying
Chromatin-modifyingenzymesenzymesprovide
provide
initial
initialcontrol
controlof
ofgene
geneexpression
expressionbybymaking
makingaa
region
regionof ofDNA
DNAmoremoreavailable
availableor
orless
less
available
availablefor
fortranscription.
transcription.

Multiple
Multiplecontrol
controlelements
elementsare
areassociated
associated
with
withmost
mosteukaryotic
eukaryoticgenes.
genes.
20
Control elements – Proximal
ProximalControl
ControlElements
Elements
→→Elements
Elementsthat
thatare
arefound
usually activators CLOSE
CLOSEtotothe
thegene
gene
found

Distal
DistalControl
ControlElements
Elements→ →
Elements
Elementsthat
thatare
arefound
found
further
furtheraway
awayfrom
fromthe
thegene,
gene,
and
andcome
comeinto
intocontact
contactwhen
when
the
theDNA
DNAbends
bends

Both
Bothofofthese
thesecancanact
actas
as
activators,
activators,which
which“grab”
“grab”
additional
additionaltranscription
transcription
factors
factorsand
andaddaddthem
them
(increases
(increasesefficiency);
efficiency);
sometimes,
sometimes,however,
however,they they
can
canact
actasasrepressors
repressorsby by
grabbing
grabbingother
othertypes
typesofof
specific
specifictranscription
transcriptionfactors
factors

21
Post transcriptional regulation – alternative RNA Splicing

Regulatory
Regulatorymechanisms
mechanisms The
Thelife
lifespan
spanofofan
anmRNA
mRNAmolecule
moleculeisisan
an
that
thatoperate
operateAFTER
AFTER important
importantfactor
factorin
indetermining
determiningthe
thepattern
patternof
of
transcription
transcriptionallow
allowaacell
cell protein
proteinsynthesis.
synthesis.
to
torapidly
rapidlyfine-tune
fine-tunegene
gene
expression
expressionininresponse
responseto to Prokaryotic
ProkaryoticmRNA
mRNAmolecules
moleculesarearetypically
typically
environmental
environmentalchanges,
changes, degraded
degradedafter
afteronly
onlyaafew
fewminutes,
minutes,while
while
without
withoutaltering
alteringits
its eukaryotic
eukaryoticmRNAs
mRNAstypically
typicallylast
lastfor
forhours,
hours,days,
days,or
or
transcriptional
transcriptionalpatterns.
patterns. weeks.
weeks.
--Alternate
AlternateRNARNASplicing
Splicing(exon
(exonshuffling)
shuffling)
this
thissignificantly
significantlyexpands
expandsthe
therepertoire
repertoire
of
ofaaset
setof
ofgenes;
genes;even
eventhough
thoughwewehave
haveaa
set
setnumber
numberof ofprotein-encoding
protein-encodinggenes…
genes…
but
butshuffling
shufflingthetheintrons/exons
introns/exonswewecan
canget
get
aamuch
muchhigher
highernumber
numberofofactual
actualproteins
proteins
--Regulating
RegulatingmRNA
mRNAdegradation
degradation
--Translational
Translationalcontrol
control(blocking
(blockinginitiation
initiation
stage
stageofoftranslation;
translation;block
blockribosome
ribosome
attachment)
attachment)
22
 Translational Regulation
 The
Theinitiation
initiationofoftranslation
translationofofan
anmRNA
mRNAcan canbe
beblocked
blockedbybyregulatory
regulatory
proteins
proteinsthat
thatbind
bindtotospecific
specificsequences
sequenceswithin
withinthe
themRNA,
mRNA,preventing
preventing
ribosome
ribosomeattachment.
attachment.
 Translation
Translationofofall
allthe
themRNAs
mRNAsin inaaeukaryotic
eukaryoticcell
cellmay
maybeberegulated
regulated
simultaneously
simultaneouslyby bythe
theactivation
activationororinactivation
inactivationofofthe
theprotein
proteinfactors
factors
required
requiredtotoinitiate
initiatetranslation.
translation.
 Post translational regulation – selective degradation

--Proteins
Proteinscan
canalso
alsobe
bemodified
modifiedafter
aftertranslation
translation(adding/
(adding/removing:
removing:phosphate
phosphate
groups,
groups,carbohydrate
carbohydrateportions,
portions,sections
sectionsof
ofAA’s)
AA’s)for
forthem
themto
tobe
befunctional
functional
--Proteins
Proteinsalso
alsoneed
needtotobe
bemoved
movedto
todifferent
differentparts
partsof
ofthe
thecell
cell(or
(orof
ofthe
theorganism)
organism)in
in
order
ordertotobe
beeffective
effective

--The
Thelength
lengthofoftime
timeaaprotein
proteinfunctions
functionsbefore
beforeititisisdegraded
degradedisisstrictly
strictlyregulated
regulated
(eg.
(eg.cyclins).
cyclins). To
Tomark
markaaprotein
proteinforfordestruction,
destruction,the
thecell
cellattaches
attachesaasmall
smallprotein
protein
called ubiquitin to it. This is called:
called ubiquitin to it. This is called:
-- Selective
Selectivedegradation
degradation→ →tagged
taggedbybyubiquitin
ubiquitinand andrecognized
recognizedbybyproteasomes
proteasomes
to
tobe
bebroken
brokendown
down 24
DIFFERENTIAL GENE EXPRESSION:
This leads to different CELL TYPES in a
multicellular organism
 In
Inthe
thedevelopment
developmentof ofmost
most
multicellular
multicellularorganisms,
organisms,aa
single-celled
single-celledzygote
zygotegives
gives
rise
riseto
tocells
cellsof
ofmany
many
different
differenttypes.
types.
 As
Asaazygote
zygotedevelops
developsintointo
an
anadult
adultorganism,
organism,its its
transformation
transformationresults
resultsfrom
from
three
threeinterrelated
interrelatedprocesses:
processes:
cell
celldivision,
division,cell
cell
differentiation,
differentiation,and and
morphogenesis.
morphogenesis.
 Cell Division, Cell Differentiation and
Morphogenesis
During
Duringdevelopment,
development,cellscellsbecome
becomespecialized
specializedinin
structure
structureand
andfunction,
function,undergoing
undergoingcell cell
differentiation.
differentiation.Different
Differentkinds
kindsofofcells
cellsare
are
organized
organizedinto
intotissues
tissuesand
andorgans.
organs. Plants
Plantscan
canbe
be
cloned
clonedfrom
fromsomatic
somaticcells
cells(that
(thathave
havealready
already
differentiated),
differentiated),so sothis
thisshows
showsthat
thatdifferentiated
differentiated
cells
cellsretain
retainall
allthe
thegenes
genesofofthe
thezygote
zygoteeven
even
though
thoughthey
theyare
arespecialized.
specialized.

Through
Throughaasuccession
successionof of The
Thephysical
physical
mitotic
mitoticcell
celldivisions,
divisions,thethe processes
processesthat
thatgive
givean
an
zygote
zygotegives
givesrise
riseto
tomany
many organism
organismitsitsshape
shape
cells.
cells. Cell
Celldivision
divisionalone
alone constitute
constitute
would
wouldproduce
produceonlyonlyaa morphogenesis,
morphogenesis,the the
great
greatball
ballof
ofidentical
identicalcells.
cells. “creation
“creationof
ofform.”
form.”
Cytoplasmic
Determinants
Maternal
Maternal
substances
substancesthat
that
influence
influencethe
the
course
courseofofearly
early
development
developmentare are
called
called
cytoplasmic
cytoplasmic
determinants.
determinants.
These
Thesesubstances
substances
regulate
regulatethe
the
expression
expressionof of
genes
genesthat
thataffect
affect
the
thedevelopmental
developmental
fate
fateof
ofthe
thecell.
cell.
 Differentiation DIFFERENTIATION
DIFFERENTIATION
isiswhen
whenaacell
cell
You
You need
needaaspecific
specificcombination
combinationofofseveral
severalregulatory
regulatoryproteins
proteins expresses
expressesgenes
genesthat
that
in
inorder
orderto
tosuccessfully
successfullydifferentiate.
differentiate. ItItisishard
hardto
torecreate
recreatethe
the encode
encodeproteins
proteinsforfor
exact
exactenvironment.
environment. that
thatspecific
specifictissue.
tissue.
Before
Beforedifferentiation
differentiation
occurs,
occurs,
DETERMINATION
DETERMINATION
occurs.
occurs. This
Thisisiswhen
when
changes
changesat atthe
the
molecular
molecularlevel
levelputputaa
cell
cellon
onaapath
pathto to
specialization.
specialization.
Embryonic
EmbryonicPrecursor
PrecursorCell
Cell

Determination

Once
Onceitithas
hasundergone
undergonedetermination,
determination,an anembryonic
embryoniccellcellisis Myoblast
Myoblast
irreversibly
irreversiblycommitted
committedto toits
itsfinal
finalfate.
fate. IfIfaadetermined
determined
cell
cellisisexperimentally
experimentallyplaced
placedin inanother
anotherlocation
locationin inthe
the Differentiation
embryo,
embryo,ititwill
willdifferentiate
differentiateasasififititwere
werein inits
itsoriginal
original
position. Muscle
MuscleCell
Cell
position. 28
 Pattern Formation
Pattern
Patternformation
formationisisthethedevelopment
developmentof ofspatial
spatial
organization.
organization. ItItdetermines
determinesthetheanimals
animals“basic
“basic
body
bodyplan”.
plan”. ItItmakes
makesvarious
varioustissues
tissuesand
and
organs
organsdevelop
developin incertain
certainplaces.
places.Pattern
Pattern
formation
formationbegins
beginsin inthe
theearly
earlyembryo,
embryo,when
whenthethe
major
majoraxes
axesof
ofanananimal
animalareareestablished.
established. Before
Before
specialized
specializedtissues
tissuesandandorgans
organsform,
form,the
therelative
relative
positions
positionsof
ofan
ananimal’s
animal’sbody
bodysymmetry
symmetry
(anterior-posterior,
(anterior-posterior,dorsal-ventral,
dorsal-ventral,right-left)
right-left)are
are
established.
established.

Similar
Similarto
tolaying
layingout
outall
allthe
theparts
partsof
ofaamodel
model
airplane
airplanein
inthe
theapproximate
approximatespotsspotsthey
theyare
are
going
goingto
togo
gobefore
beforeyou
youput
putitittogether.
together.
In
Inanimals,
animals,pattern
patternformation
formationoccurs
occursduring
duringthe
the
embryo
embryoand
andjuvenile
juvenilestages.
stages.

In
Inplants,
plants,pattern
patternformation
formationoccurs
occursthroughout
throughout
the
thelife
lifeof
ofthe
theplant
plantbecause
becausethey
theyhave
haveapical
apical
meristems.
meristems. 29
 Studies
Studiesofofpattern
patternformation
formationhave
Homeotic Genes established
establishedthat
thatgenes
genescontrol
control
have

development
developmentand andhave
haveidentified
identifiedthe
thekey
key
roles
rolesof
ofspecific
specificmolecules
moleculesin indefining
defining
position
positionand
anddirecting
directingdifferentiation.
differentiation.
These
Thesegenes
genesare
arecalled
calledhomeotic
homeoticgenes
genes
and
andwere
werefound
foundtotobe
behighly
highlyconserved
conservedinin
evolution.
evolution. Changes
Changesin inthese
thesegenes
genescan
can
lead
leadto
totransformations
transformationsin inentire
entirebody
body
parts.
parts.
Homeotic
Homeoticgenes
genesare
areconsidered
consideredto tobe
bethe
the
MASTER
MASTERREGULATORY
REGULATORYGENES. GENES. TheyThey
encode
encodetranscription
transcriptionfactors
factorsthat
thatcan
can
control
controlthe
theexpression
expressionofofother
othergenes,
genes,
especially
especiallygenes
genesfor
foranatomical
anatomicalfeatures.
features.

30
 Maternal Effect Gene
(in fruit flies)

 AAmaternal
maternaleffect
effectgene
geneisisaagene
genethat,
that,when
whenmutant
mutantininthe
themother
mother(in(inDrosophila),
Drosophila),
results
results in a mutant phenotype in the offspring, regardless of the offspring’sown
in a mutant phenotype in the offspring, regardless of the offspring’s own
genotype.
genotype.
 Maternal effect genes are also called egg-polarity genes because they control the
 Maternal effect genes are also called egg-polarity genes because they control the
orientation
orientationofofthe
theegg
eggand
andconsequently
consequentlythethefly.
fly.
 One group of genes sets up the anterior-posterior axis, while a second group establishes
 One group of genes sets up the anterior-posterior axis, while a second group establishes
the
thedorsal-ventral
dorsal-ventralaxis.
axis.

Bicoid Gene
 One
Oneexample
exampleofofaamaternal
maternaleffect
effectgene
geneisiscalled
calledaabicoid
bicoidgene
geneaffects
affectsthe
thefront
fronthalf
half
of
ofthe
thebody
body(anterior/posterior
(anterior/posterioraxis).
axis).
 An
Anembryo
embryowhose
whosemother
motherhashasaamutant
mutantbicoid
bicoidgene
genelacks
lacksthe
thefront
fronthalf
halfofofits
itsbody
body
and
andhas
hasduplicate
duplicateposterior
posteriorstructures
structuresat atboth
bothends.
ends.
 This
Thissuggests
suggeststhat
thatthe
theproduct
productof ofthe
themother’s
mother’sbicoid
bicoidgene
geneisisessential
essentialfor
forsetting
settingup
up
the
theanterior
anteriorend
endof
ofthe
thefly
flyand
andmight
mightbe beconcentrated
concentratedat atthe
thefuture
futureanterior
anteriorend.
end.
 Cancer
 Cancer
Cancerisisaaset
setof
ofdiseases
diseasesin inwhich
whichcells
cellsescape
escapethe
thecontrol
controlmechanisms
mechanismsthat
that
normally
normallyregulate
regulatecell
cellgrowth
growthandanddivision.
division.
 The
Thegenes
genesthat
thatnormally
normallyregulate
regulatecell
cellgrowth
growthandanddivision
divisionduring
duringthe
thecell
cellcycle
cycle
include
includegenes
genesfor
forgrowth
growthfactors,
factors,their
theirreceptors,
receptors,and
andthe
theintracellular
intracellularmolecules
molecules
of
ofsignaling
signalingpathways.
pathways.
 Mutations
Mutationsaltering
alteringany
anyofofthese
thesegenes
genesin insomatic
somaticcells
cellscan
canlead
leadto
tocancer.
cancer.
 The
Theagent
agentofofsuch
suchchanges
changescan canbe
berandom
randomspontaneous
spontaneousmutations
mutationsor or
environmental
environmentalinfluences
influencessuchsuchas
aschemical
chemicalcarcinogens,
carcinogens,X-rays,
X-rays,and
andsome
someviruses.
viruses.
 Proto-
oncogenes vs.
Oncogenes

Proto-oncogenes
Proto-oncogenes→ →normal
normalgenes
genesthat
thatmake
make
enzymes
enzymesthat
thatregulate
regulatethe
thecell
cellcycle
cycle
Oncogenes
Oncogenes→ →mutated
mutatedproto-oncogenes;
proto-oncogenes;
can
canlead
leadto
tocancer
cancer

AAproto-oncogene
proto-oncogenebecomes
becomesananoncogene
oncogene
following
followinggenetic
geneticchanges
changesthat
thatlead
leadtotoan
an
increase
increasein
inthe
theproto-oncogene’s
proto-oncogene’sprotein
protein
production
productionororin
inthe
theintrinsic
intrinsicactivity
activityof
of
each
eachprotein
proteinmolecule.
molecule.
33
The
Thenormal
normalproducts
productsofoftumor-
tumor-
suppressor
suppressorgenes
genesinhibit
inhibitcell
celldivision
division
by
byencoding
encodingproteins
proteinsthat
thathelp
helpprevent
uncontrolled
prevent Tumor-Suppressor Genes
uncontrolledcell
cellgrowth.
growth.

Some
Sometumor-suppressor
tumor-suppressorproteins
proteins
normally
normallyrepair
repairdamaged
damagedDNA,
DNA, The
preventing TheRas
Rasprotein,
protein,thetheproduct
productof ofthe
theras
ras
preventingthe
theaccumulation
accumulationof
ofcancer-
cancer- proto-oncogene,
proto-oncogene,isisaaGGprotein
proteinthat
thatrelays
relays
causing
causingmutations.
mutations. aagrowth
growthsignal
signalfrom
fromaagrowth
growthfactor
factor
receptor
receptorononthe
theplasma
plasmamembrane
membraneto toaa
Mutations
Mutationsininthe
theproducts
productsof
oftwo
twokey
key cascade
genes, cascadeofofprotein
proteinkinases
kinases this
this
genes,the
theras
rasproto-oncogene
proto-oncogeneandandthe
the stimulates
stimulatesthe
thecell
cellcycle!
cycle!AAmutation
mutationin in
p53
p53tumor-suppressor
tumor-suppressorgene,
gene,occur
occurin
in this
30% thiscan
cancause
causethe
thecell
cellcycle
cycleto
tobe
be
30%and
andover
over50%
50%ofofhuman
humancancers,
cancers, constantly
constantlyturned
turnedON.
ON.
respectively.
respectively.
 “Guardian
“GuardianAngel
Angelof ofthe
theGenome”;
Genome”;functions
functionsas asaa
transcription
transcriptionfactor
factorand
andactivates
activatesthe
thep21
p21gene
gene(which
P53 genes creates
createsaaproduct
productthat
thathalts
haltsthe
thecell
cellcycle
cycleto
toleave
(which
leavetime
time
for
forDNA
DNAto torepair
repairitself)
itself)

Defective
Defectivep53
p53==no
no
active
activep21
p21==no
nohalting
halting
the
thecell
cellcycle
cycle

p53
p53 
-- Slows
Slowscell
cellcycle
cycle
-- Causes
Causesapoptosis
apoptosis(cell
(cell
suicide)
suicide)
-- Acts
Actsasasaa
transcription
transcriptionfactor
factor
for
forp21
p21
-- Prevents
Preventscells
cellsfrom
from
passing
passingon onmutations
mutations
in
indamaged
damagedDNA DNA
-- IsIsan
anexample
exampleof ofaa
tumor
tumorsuppressor
suppressor
gene
gene
35
 Cancer
 More
Morethan
thanone
onesomatic
somaticmutation
mutationisisgenerally
generallyneeded
neededtotoproduce
producethe thechanges
changes
characteristic
characteristicof ofaafull-fledged
full-fledgedcancer
cancercell.
cell.
 Typically you need to have several oncogenes and mutations in multiple
Typically you need to have several oncogenes and mutations in multiple
tumor-suppressor
tumor-suppressorgenes.genes.
 IfIfcancer
cancerresults
resultsfrom
froman anaccumulation
accumulationof ofmutations,
mutations,and
andififmutations
mutationsoccur
occur
throughout
throughoutlife,
life,then
thenthe
thelonger
longerwe
welive,
live,the
themore
morelikely
likelywe
weareareto
todevelop
develop
cancer.
cancer.
 Cancer Genetic Linkage

 The
Thefact
factthat
thatmultiple
multiplegenetic
geneticchanges
changesarearerequired
requiredto toproduce
produceaacancercancercell
cellhelps
helps
explain the predispositions to cancer that run in families.
explain the predispositions to cancer that run in families.

 An
Anindividual
individualinheriting
inheritingan anoncogene
oncogeneor oraamutant
mutantallele
alleleof
ofaatumor-suppressor
tumor-suppressor
gene
geneisisone
onestep
stepcloser
closerto
toaccumulating
accumulatingthe thenecessary
necessarymutations
mutationsfor forcancer
cancerto to
develop.
develop.

 Geneticists
Geneticistsare
aredevoting
devotingmuch
mucheffort
effortto
tofinding
findinginherited
inheritedcancer
canceralleles
allelesso
sothat
thataa
predisposition
predispositionto tocertain
certaincancers
cancerscancanbebedetected
detectedearly
earlyininlife.
life.

 Mutations
Mutationsin inone
onegene,
gene,BRCA1,
BRCA1,increase
increasethetherisk
riskofofbreast
breastand
andovarian
ovariancancer.
cancer.

 Mutations
Mutationsin inBRCA1
BRCA1and andthe
therelated
relatedgene
geneBRCA2
BRCA2are arefound
foundin inatatleast
leasthalf
halfofof
inherited
inheritedbreast
breastcancers.
cancers.

 Both
BothBRCA1
BRCA1and andBRCA2
BRCA2areareconsidered
consideredtumor-suppressor
tumor-suppressorgenes genesbecause
becausetheir
their
wild-type alleles protect against breast cancer and their mutant
wild-type alleles protect against breast cancer and their mutant alleles are alleles are
recessive.
recessive.