control over genes
Post on 18-Feb-2016
247 Views
Preview:
DESCRIPTION
TRANSCRIPT
LIF 101 16th October 2016
MaleFemale
Mating
Parents &Children:1 boy; 1 girl(in order of birth)Dizygotic(non identical Twins)
Monozygotic(identicalTwins)
SexunspecifiedAffectedindividuals
Heterozygotesfor autosomal recessiveCarrier of sex linked recessive
Death
Abortion or still birth
Propositus
Consanguineousmarriage
Symbols used in pedigree analysis:RECAP
Pedigree Analysis
Normal female Normal maleMarriage
1st born
Siblings
Affected
I
II
One these parent was therefore a
carrier
Is the mutant allele dominant or recessive?
RECAP
Albinism : Autosomal recessive inheritance
Why this inheritance is called autosomal?
RECAP
Achondroplasia: Autosomal dominant inheritence
Why this called autosomal dominant
dominant ?
RECAP
Sex Determination in humans
Parents
RECAP
Female carrier* mates with normal male
FXNXN
Y
XN
XA
• Half* her daughters will be carriers• Half* her sons will be affected
F x MXNXA XNY
Eggs
Sperm
XN
MXNY
normal normal
MXAY
affected
FXAXNcarrier
*on average
X-linked recessive diseaseRECAP
Pedigree Illustrating X-Linked Recessive Inheritance Pattern
Grand daughter
Uncle (mama)
Grand Uncle
Nephew
Great grand mother
Grand motherNani
Hemophilia : X-linked recessive
Queen Victoria (1819-1901) passed haemophilia A
on to many of her descendants
Empress of India 1876
Haemophilia leads to a severely increased risk of prolonged bleeding from common injuries, or in severe cases bleeds may be spontaneous and without obvious cause.
Haemophilia A is a genetic deficiency in clotting factor VIII which causes increased bleeding and usually affects males. About 70% of the time it is inherited as an X-linked recessive trait, but around 30% of cases arise from spontaneous mutations.
Victoria (Princess of Saxe-Coburg)
Edward(Duke of Kent)
Victoria (Queen of England) Albert
Victoria Frederick
Wihelm II Sophie George V
George VI
Queen Elizabeth
Prince Philip
Alice Louis Alfred Helena LouiseArthur
Leopold Helen Beatrice Henry
Nikolas II of RussiaAlix
Waldemar Henry
Alfonso XII Spain Eugenic
Alfonso Gonzalo
Alexic
Normal Male
Normal Female
Hemophilic Male
Carrier Female
Male died in infancyPossible hemophilic?
Inheritance of eye coloration in fruit fly, Drosophila:An example of sex-linked inheritance
Normal Red Mutant white
12
Then test and infer w w
w+w
W+ w+w+
w+w w+w
W
w
Problem TEST: A yellow body-colored female fruit fly was
mated to a male with normal body color (brown).
RESULT: All the female progeny were brown (normal) while all the male progeny were yellow.
EXPLAIN: What does this inheritance pattern signify?
Display this cross
What if individual chromosomes do not separate during meiosis? Error in chromosome number
Significance of chromosome numbers in in heritance
The Culprit: Non-disjunction
Older women are more likely than younger women to evidence chromosome damage and meiotic
irregularities
Pre-natal Diagnosis: Amniocentesis
Fetal cells present in amniotic fluid are then cultured and analyzed for chromosomes (karyotyping) and other biochemical tests
Will it be possible to detect a defective
gene by amniocentesis?
Monosomy: Turner’s Syndrome: 44+X
This baby has one X
chromosome. Yet it is a
female baby. Why?
Individual chromosome segregation going wrong during meiosis
Trisomy of chromosome . 21: Down’s Syndrome: 45+XY/45+XX
Other disorders: Trisomy-13 (47:+13), Trisomy-18 (47:+18), Klinefelter Syndrome (44+XXY), Triple X syndrome (44+XXX), XYY
trisomy (44+XYY)
Why down’s syndrome does
not suggest abnormal sex
linked inheritance?
How can errors in chromosomal
segregation result in Down’s
syndrome?
People suffering from Down syndrome can lead a reasonably normal life
Genes and, chromosomes and some interesting facts about our inheritance
These cats are known as ‘Calico cats’ marked by their patchy coat (fur) coloration . The
coloration gene (or fur pigmentation gene) is present on the X chromosome (X-linked)
Coat coloration in XB/Xb cats
B= orange alleleb= Black allele
Possible Female genotypes
Possible male genotypes
Female cell where X carrying orange
allele is inactivated
Female cell where X chromosome carrying black allele allele is inactivated
However, one of the X chromosomes is randomly inactivated in female cat body
cells
Explain why male cats
have uniform coat
color
Explain why these female cats have uniform coat color
Explain why these female cats have patchy coat color
Note the fur/coat coloration in male and female cats
How will you name the gene that regulate eye
coloration?
How will you decide which one is recessive
and which one is dominant ?
What will be the consequence if the while gene is X-linked and Drosophila female and male have XX and XY sex chromosome constitution?
What types and proportions of F2 progeny are expected the white
gene were located on an autosome?
P1
Predict the phenotypic ratios in the progeny if a Drosophila female, heterozygous for white mutation is mated with a white eye-colored male ?
Do you think inheritance of white gene in Drosophila follows Mendelian laws?
Do you think inheritance of white gene display a chromosomal basis of inheritance? Which chromosome?
What if individual chromosomes do not separate during meiosis? Error in chromosome number
What if entire sets of chromosome do not separate? Error in chromosome sets
Significance of chromosome numbers and sets
Polyploidy in plants Colchicine inhibits chromosome segregation
In nature 3n, 4n, 6n and 8n plants are very common
Many varieties of wheat, cotton, banana, tobacco , strawberries etc are polyploids
Polyploidy results in bigger grain or fruit sizes and high yield which is beneficial
Thus, polyploidy is also artificially induced in plants by breeders using colchicine.
Polyploidy in plants
The cultivated strawberry is an octaploid hybrid derived from hybridization of two
natural octaploids.
Potato flowers. Potato is an autotetraploid.
This supermarket banana is most likely the common variety Cavendish, an autotriploid of Musa acuminata
Seeds of bread wheat. Wheat is a allohexaploid. Its genome consists of three ancestral parental genomes
DNA as a genetic material
Griffith’s experiment 1927:discovery of transforming factor (TF))
Non-virulent strain does
not kill
Heating kills virulent strain
Heat-killed virulent strain can transfer its
virulence “factor” to a non-virulent
strain
First historical demonstration of an chemical entity for genetic inheritance
Dies
Dies
Lives
Lives
Virulent
Non-virulent
Virulent, heat killed
Non-virulent + heat killed
Virulent strain kills the host
Pneumococcal infection
What does this heat killing signify with respective genetic inheritance
What is genetic transformation?
Which one was genetically transformed? Mouse or the bacteria?
Avery, MacLeod, McCarty
• Purified the transforming factor (TF) to investigate its properties.
• Salient points:– Treatment with protease did not destroy
TF– Treatment with RNase did not destroy TF– Purified TF did not have sugars
• Thus TF should be DNA Do these terms transforming factors, Mendelian factors and
DNA carry similar meaning ?
What about Mendelian alleles? Are these too transforming factors, Mendelian factors?
What is there in transforming or Mendelian factor? And now in DNA?
DNA carries digitized information Four digits are used to write genetic
codes
Watson and Crick proposed structure of DNA 1953
Thus, to understand the nature of genetic information we need to understand the basic DNA structure and how information flows:
Within an individual.
From one generation to the other
Within a cell
Journal Nature in its 171st volume on pages 737–738 ( 25 April 1953)
DNA is a very large molecule made up of a long chain of sub-units
The sub-units are called nucleotides
Each nucleotide is made up of
a sugar called deoxyribose
a phosphate group -PO4 and
an organic base
DNA molecule RECAP
A few general features you need to remember about DNA structure
Chargaff’s RuleChargaff’s Rule
• AdenineAdenine must pair with ThymineThymine
• GuanineGuanine must pair with CytosineCytosine
• Their amounts in a given DNA molecule will be about the sameabout the same.
G CT A
RECAP
DNA Double HelixDNA Double Helix
P
P
P
O
O
O
1
23
4
5
5
3
3
5
P
P
PO
O
O
1
2 3
4
5
5
3
5
3
G C
T A
H-bonds
H-bonds
RECAP
´
DNA strands run anti-parellel
´
Mark the features of DNA that makes it ideal as a genetic
materials
Sequence in one strand serves as the
template for the sequence in the
other
Sequence of nucleotides in DNA strands could store
genetic information
RECAP
DNA RNA protein Transcription Translation
Replication
Reverse transcription
Central dogma of Molecular biology सि�द्धांत
In subsequent lectures we will explore this flow of
information the decoding process
Flow of information
Nucleic acid to protein
Flow of information: nucleic acid to nucleic
acid
Nature of information flow
Why there is no loss of
information during
replication? How much of the information in DNA goes to
RNA??
Replication: synthesis of daughter DNA from parental DNA Transcription: synthesis of RNA using DNA as the template Translation: protein synthesis using mRNA molecules as the
template Reverse transcription: synthesis of DNA using RNA as the template
Flow of genetic information
Test your understanding
How many DNA molecules are present in a metaphase chromosome?
How many DNA molecules are present in a chromatid?
Are the DNA sequences of two sister chromatids are identical?
How many DNA molecules are there in a human sperm?
Question
Which aspect of information flow a sperm represent ?
Which aspect of information flow does these sister chromatids
symbolize ?
Approximate gene numbersChromosome 1 : 4220Chromosome 2 : 1,491
Chromosome 3 : 1,550
RECAP
Why did the author say that human genome is written in 23 chapters?
Why not 46?
Does a cell read these chapters one
by one - in a sequence?
How many chapters are there in our book of life?? And how many words/sentences are there in each chapter??
What is implied when one says that genetic information is being
readout?
Here ‘book of life’ is a metaphor
लाक्षणि�क रूप �ेउपमा
AUTOBIOGRAPHY OF A SPECIES IN 23 CHAPTERS
What is a gene/ Mendelian factor:
a letter, a word or
a sentence?
Next lecture:We would examine the nature of genetic information and how it flows within a cell
Slide series 6
Flow of genetic information: Three aspects -
Transcription, Translation&
Genetic code
Let’s first appreciate the challenge of transfer of information from DNA
(genes) to protein
Consider the case of Sickle Cell Anemia?
A serious condition in which red blood cells can become sickle-shaped
Normal red blood cells are smooth and round. They move easily through blood vessels to carry oxygen to all parts of the body.
Sickle-shaped cells don’t move easily through blood. They’re stiff and sticky and tend to form clumps and get stuck in blood vessels.
The clumps of sickle cell block blood flow in the blood vessels that lead to the limbs and organs. Blocked blood vessel can cause pain, serious infection, and organ damage.
While the defect is seen in the protein, sickle cell anemia is produced by changes in gene sequence
Normal and Sickled Red Blood Cells in Blood Vessels
Figure A shows normal red blood cells flowing freely in a blood vessel. The inset image shows a cross-section of a normal red blood cell with normal hemoglobin.
Figure B shows abnormal, sickled red blood cells clumping and blocking the blood flow in a blood vessel. The inset image shows a cross-section of a sickled red blood cell with abnormal strands of hemoglobin.
Source from http://www.nhlbi.nih.gov/health/dci/Diseases/Sca/SCA_WhatIs.html
• What could be the reason(s) for the wrong message from the DNA that produced a wrong protein (hemoglobin)?
• How ‘information’ encoded in the DNA of the nucleus is transferred to the cytoplasm
• How does this information ‘translate’ into protein??
Questions?
DNA RNA protein Transcription Translation
Replication
Reverse transcription
Central dogma of Molecular biology सि�द्धांत
We will now explore this flow of information: that is,
process of decoding the genetic message
Flow of information
Nucleic acid to protein
Flow of information: nucleic acid to nucleic
acid
Nature of information flow
Why there is no loss of
information during
replication? How much of the information in DNA goes to
RNA??
RECAP
Flow of genetic information A gene is transcribed into RNA
which is then translated into the polypeptide
DNA
Transcription
RNA
Protein
Translation
FLOW OF GENETIC INFORMATION FROM DNA TO RNA TO PROTEIN
Gene (DNA) is a linear sequence of many nucleotides DNA is transcribed into a linear sequence
of RNA RNA is translated into a linear sequence of
amino acids – polypeptides (protein)
Entire business of flow of genetic information thus depends on constructing (transferring information) one kind of polymer from
another
DNA RNA Protein(polypeptide chain
Transcription – Step I
Template DNA Strands unzip locally
T G C A T A G C G C A T
Transcription – Step II
A C G T A T C G C G T A U G C A U A G C G C A U
One of the DNA strands serve as a template and copied as mRNA
(messenger RNA)
mRNA
DNA
A C G T A T C G C G T A T G C A T A G C G C A T
U G C A U A G C G C A U
DNA
RNA
mRNA is then releases
Chemical structure of RNA
Notice the difference between ribose and deoxyribose in the figure above. In ribose, carbon atom #2 carries one hydroxyl group (colored red). In deoxyribose, carbon atom #2 carries a hyrogen atom instead of a hydroxyl group.
Cytosine
Thymine
Uracil
Pyrimidine bases
DNA/RNA DNA RNA
In place of thymine in DNA
Transcription
RNA polymerase RNA nucleotides
Direction of transcription Template
Strand of DNANewly made RNA
TC
A T C C A A TT
GG
CC
AATTGGAT
G
U
C A U C C A AU
In the nucleus, the DNA helix unzips And RNA nucleotides line up along
one strand of the DNA, following the base pairing rules
As the single-stranded messenger RNA (mRNA) peels away from the gene
The DNA strands rejoin
Steps in transcription of a geneRNA polymerase
DNA of gene
PromoterDNA Terminator
DNA
Area shownIn Figure 10.9A
GrowingRNA
Completed RNARNApolymerase
1 Initiation
2 Elongation
3 Termination
T A C T T C A A A A T C
A T G A A G T T T T A G
A U G A A G U U U U A G
Transcription
Translation
RNA
DNA
Met Lys PhePolypeptide
Startcondon
Stopcondon
Strand to be transcribed
How does DNA/RNA code for protein?
Codon - a sequence of nucleotides that
codes for an amino acid
STEP III TRANSLATION
How can four nucleotides code for 20
amino acids? If one nucleotide codes for one amino acid then a maximum of four amino acid could be coded
If two nucleotide code for one amino acid then a maximum of 16 amino acid could be coded
If three nucleotides code for one amino acid then a maximum of 64 codons can be generated, while the number of amino acids are only 20
The dictionary of genetic code (64 potential codons)
आनुवान्शि�की कोड की शब्दकोश 5’-3’
A dictionary
offers translation from one language
to the other. Which two
languages are being translated
in a genetic
dictionary?
Why do we need
this translation
?
Where does it take
place?
What is the name
of this process?
Hallmarks of genetic code
Triplet : three consecutive nucleotides code of one amino
acid
Redundant : meaning not unique (there are more than one codon for most amino acids)
Universal: All organisms bacteria to human use the same genetic codes/dictionary
Comma-less: meaning consecutive codons are readout into an amino acid sequence without gaps
How does an universal genetic code impact Biotechnology??
What would have been the case if bacterial genetic code were to be different from human?
What do we understand when we mean when we say that genetic code is universal?
DNA
Messenger-RNA
Transfer RNA
Amino acid
Key players in decoding the genetic message
Transfer RNA
Leucine
Codon
Anti-codon
How genetic code is readout in the cell
A C G T A T C G C G T A T G C A T A G C G C A T
DNA
3’ U G C A U A G C G C A U 5’ mRNA
5’
3’
5’G U C
3’
C A G
3’5’
tRNA
Amino acid
Anti-Codon
Codon
Many amino acids are specified by more than one codon-degeneracy
Codons specifying the same amino acid are called synonyms
Connect the three points about genetic codes: redundancy, degeneracy and synonymous
CUG CUC
Codon-anticodon pairing of two tRNA Leu molecules
5’- 3’ 5’- 3’Codon
GAC3’- 5’
Anti- codon GAG3’- 5’
Three codons direct chain termination
Three codons, UAA, UAG, and UGA signify chain termination.
They are not read by tRNAs but by proteins called release factors (RF1 and RF2 in bacteria and eRF1 in eukaryotes).
Three Rules
Codons are read in a 5’ to 3’ direction. Codons are non-overlapping and the
message contains no gaps. The message is translated in a fixed
reading frame which is set by the initiation codon.
FATCATATETHERAT
Meaning and consequences of non-overlapping & comma-less reading of the triplet codons
FATATATETHERAT
Let’s consider a metaphor of a normal
DNA sequence
What is the
problem here?
What is the
consequence?
Mutation: a change in genetic code
1. Missense mutation: An alternation that changes a codon specific for one amino acid to a codon specific for another amino acid.
2. Nonsense or stop mutation: An alternation causing a change to a chain-termination codon.
3. Frame-shift mutation: Insertions or deletions of one or a small number of base pairs that alter the reading frame.
How the genetic code was cracked?
The use of artificial mRNAs and the availability of cell-free systems for carrying out protein
synthesis began to make it possible to crack the code
Codon Assignments from Repeating Copolymers
Organic chemical and enzymatic techniques were
used to prepare synthetic poly-ribonucleotides with known
repeating sequences.
Experimental Results:
UUU codes for phenylalanine. CCC codes for proline. AAA codes for lysine. The guanine residues in poly-G firmly
hydrogen bond to each other and form multistranded triple helices that do not bind to ribosomes.
copolymer Codons Recognized
Amino Acids Incorporated
Codon Assignment
(CU)” CUC|UCU|CUC… Leucine 5’-CUC-3’ Serine UCU (UG)” UGU|GUG|UGU… Cystine UGU Valine GUG (AC)” ACA|CAC|ACA… Threonine ACA Histidine CAC (AG)” AGA|GAG|AGA… Arginine AGA Glutamine GAG (AUC)” AUC|AUC|AUC… Polyisoleucine 5’-AUC-3’
What Are Mutations?What Are Mutations?
• Changes in the nucleotide sequence of DNA
• May occur in somatic cells (aren’t passed to offspring), but could cause diseases, such as cancer
• May occur in gametes (eggs & sperm) and be passed to offspring
Nature of point mutations
1. Missense mutation: An alternation that changes a codon specific for one amino acid to a codon specific for another amino acid.
2. Nonsense or stop mutation: An alternation causing a change to a chain-termination codon.
3. Frameshift mutation: Insertions or deletions of one or a small number of base pairs that alter the reading frame.
Point Mutation Change of a single nucleotide Includes the deletion,
insertion, or substitution of ONE nucleotide in a gene
Types of Gene/Point Mutations
Substitutions InsertionsDeletionsFrameshift
Frameshift Mutation
Inserting or deleting one or more nucleotides
Changes the “reading frame” like changing a sentence
Proteins built incorrectly
Gene Mutation Animation
Problems THE FAT CAT ATE THE BIG RAT
THE FAT ATA TET HEB IGR AT Delete C
THE FAT ATA ATE THE BIG RAT Insert A
Frame shift mutation
RBC Structure
of Hb
Primary sequence of Hb –first 26 amino acid
RECALL
Example Point Mutation
• Sickle Cell disease is the result of one nucleotide substitution
• Occurs in the hemoglobin gene
Problem
Before the true nature of the genetic coding process was fully understood, it was proposed that the message might be read in overlapping triplets. For example, the sequence of GCAUC might be read as GCA CAU AUC
G C A U C
Think of test that would negate this possibility
Consider a gene that specified the structure of hemoglobin. Arrange the following events in the most likely sequence in which they would take place
a. Anemia is observed
b. The shape of the oxygen binding site is altered
c. An incorrect codon is transcribed into hemoglobin mRNA
d. The ovum (female gamete) receive a high radiation dose
e. An incorrect codon is generated in the DNA of the hemoglobin gene
f. A mother (an X-ray technician) accidentally steps in front of an operating X-ray generator
g. A child dies h. The oxygen-transport capacity of the body is severely impairedi. A wrong tRNA is attached j. Nucleotide pair substitution occurs in the DNA of the gene for hemoglobin
If an mRNA of human beta-globin gene is added to bacterial cell free extract then human beta globin polypeptide is formed (translated)What does this observation signify
with respect to genetic code?
Question
Home assignment The amino acid sequence shown in the following table was obtained from the central region of a particular polypeptide chain in the wild type and several mutant bacterial strains
a. Wild type phe leu pro thr val thr thr arg trp b. Mutant 1 phe leu his his gly asp asp thr val c. Mutant 2 phe leu pro thr met thr thr arg trp
d. Mutant 3 phe leu pro thr val thr thr arg
e. Mutant 4 phe pro pro argf. Wild type phe leu pro ser val thr thr arg trp
1 2 3 4 5 6 7 8 9
Codon
For each mutant, say what change has occurred at the DNA level, whether the change is a base pair substitution or frame shift mutation, and in which codon the mutation occurred
Home assignment In mutant strain X of E. coli, a leucine tRNA that recognizes the codon 5’-CUG-3’ in normal cell has been so altered that is now recognizes the codon 5’-GUG-3’.
A missense mutation (lets call this mutation Y) that affects amino acid 10 of a particular protein is suppressed in mutant X cells (that is when the cells are mutant for both X and Y, which now appears wild type)
a. What mutational even has occurred in mutant X cells?
b. What amino acid would normally be present at position 10 of the protein (without the missense mutation)?
c. What amino acid is put in at position 10 if the missense mutations is not suppressed ?
d. What amino acid is put in at position 10 if the missense mutations is suppressed ?
Slide series 7
Gene regulation - Prokaryote- Eukaryotes
Gene regulation
Regulation of the synthesis of a gene’s transcript and its protein
product is termed as gene regulation.
Why control gene expression?
Each of our cells carry entire genetic instructions for our growth, development and metabolic functions.
Some of these genes are needed to be expressed all the time: respiration, for instance. These are also called house keeping genes
Other genes are not expressed all the time.
These are switched “on” and “off” on demand
Transcriptional Regulation is mediated by controlling the access of the RNA polymerase to
the promoter.
RNA polymerase Gene
Promoter Direction of transcription
Basic schemePromoter:
RNA Pol binding site in DNA sequence
upstream of transcription start point
The promoters of genes transcribed by RNA polymerase II consist of a core promoter and a regulatory promoter that contain
consensus sequences.
Not all the consensus sequences shown are found in all promoters.
A point to remember:Like transcription
initiation there are also transcription termination
mechanisms
We will not go in to these details either
Negative Regulation
Activator
RNA Polymerase
X
Repressor
Binding of repressor blocks the
binding of the RNA Pol to the
promoter
Positive Regulation
X
RNA Polymerase
An activator help RNA Pol bind to the promoter
Two broad strategies for regulation of prokaryotic transcription
In prokaryotes gene regulation takes place in response to the environment
E. coli can use either glucose, which is a monosaccharide, or lactose, which is a disaccharide
However, lactose needs to be hydrolysed (digested) first
So the bacterium prefers to use glucose when it is available.
On the other hand, when only lactose is available it turns “ON” the genes and therefore the enzymes required for lactose breakdown
Glucose
Glucose Galactose
Lactose
When lactose is absent in the medium, the lac Z gene is switched-off . That is, no mRNA is transcribed and no proteins are made.-In the presence of lactose in the medium, lac Z gene is turned on. That is, mRNA is transcribed and the proteins are made.
X
X
REPRESSOR
INDUCER
INACTIVE REPRESSOR
OPERATOR:Binding site for repressor
Regulation of the lac Z gene – an example of negative regulation ( lacZ gene codes for -galactosidase enzyme that breaks down
lactose ) What can
be the inducer of the lac Z
gene?
Answer:lactose
X
Illustration of a NEGATIVE REGULATION gene expression
INDUCIBLE TRANSCRIPTION
X
REPRESSOR ACTIVE
INDUCERREPRESSOR
INACTIVE
Regulation of lac Z gene This is also called
an inducible model of gene
regulation. WHY?
Repressor binding site
OPERATOR
Question:1Repressor is a protein: should
there be a separate gene for repressor?
Question:2How will lac Z gene be regulated if a gene coding for
repressor is mutatedQuestion:3
What will be the consequence if
operator sequence is altered?Question:4
What will happen if promoter sequence is
altered
ConceptMutations need always directly alter the target gene. There are
alternative ways of altering gene expression, other than mutations within the gene
RNA POl
Further, lac Z gene is co-regulated along with two other genes called lac A and lac Y
A set of gene co-regulated under one promoter are called operon. Thus, an operon represents a group of genes that are transcribed at the same time. They usually control an important biochemical process. They are only found in prokaryotes.
The lac Operon
P O lacZ lacY lacA
mRNA 5’ 3’
RIBSOSOME BINDING SITE
How many messenger
RNA are made by the lac operon?
P O lacZ lacY lacA
Proteinsb-galactosidase Permease Transacetylase
The lac Operon
mRNA 5’ 3’
How many polypeptide
(proteins) are made by the lac operon?
Why would you consider the lac operon a smart system?
In the presence of lactose in the medium, lactose binds to the repressor. The lactose-repressor complex is unable to bind to the
operator.
P O lacZ lacY lacA
X NO mRNA
lactose
P O lacZ lacY lacA
X NO mRNA
Regulation of the lac Operon by the repressor
Note that there are two binding sites in the repressor one for the operator and the other for
the inducer
Once “Inducer-Repressor” complex moves away from the Operator (O) RNA polymerase can bind to the
promoter (P) and initiate transcription of the lac Operon
P O lacZ lacY lacA
mRNA
Regulation of the lac Operon by the repressor
P O lacZ lacY lacA
lac repressor
X NO mRNA
lac repressorThe lac repressor binds to the operator and
inhibits transcription of the lac operon.
About the lac I gene that codes for the repressor
Question: Should the lac I gene be
always active (constitutive) or sometimes active
(inducible)?
QuestionLac repressor has two binding sites: one for binding with ‘Operator’ and the other
for the ‘Inducer’. What will be the consequence if a mutation alters its
‘inducer binding’ site
Eukaryotic Gene Regulation:
Most eukaryotic genes are controlled individually and have regulatory sequences that are much more complex than those of the lac operon.
All of the cells in a multi-cellular organism carry the complete genetic code in their nucleus, but only a few of the available genes can be expressed in the appropriate cells of different tissues.
Complex regulation allows for this specificity.
Why is gene regulation in eukaryotes more complex than in prokaryotes?
Gene regulation determined the complexities of the eukaryotic
organisms
Eukaryotic DNA bind with a variety of protein.
DNA and a group of proteins called ‘histones’ form the primary chromatin structure.
Chromatin in turn complexes with a large array of ‘non-histone’ proteins
chro
mat
inch
rom
osom
e
Incr
easin
g co
mpl
exity
of h
ighe
r ord
er st
ruct
ure
GGGCGG
CCAAT
TATA
mRNA
-200 bp -100 bp -30 bp
Promoter proximal elements Promoter
Region upstream of the transcription start site in higher eukaryotes
Promoter proximal elements
Regulatory protein
Promoter Transcribed region of a gene
RNA polymerase
Regulatory elements
Promoter
RNA transcript
DNA
Regulatory elements and promoter of an eukaryotic gene
The effect of specific point mutations in the proximal regulatory elements and promoter on β-globin gene transcription. Each line represents the transcription level of in a mutant relative to that of a wild-type β-globin gene. Red arrows shows some the mutations that affect gene transcription. The black dots represent nucleotides for which no mutation had been generated.
Rela
tive
tran
scrip
tion
leve
l Functional role of the regulatory elements and
promoter Question:Can changes
in the sequence of regulatory sequences too cause
gene mutation?
Applications of the understanding of the gene regulation
What if lacZ gene is fused with the promoter of another gene?
lacZ
A different promoter
Background information Genes expression is regulated during development – the colored stripes represent the areas where a certain gene (named even-skipped) is expressed . Note a total of total
seven stripes
Normal 7 stripes of even-skipped gene in Drosophila embryo
Stripe 2 module
Regulatory elements of a gene determine its expression in specific locations
Now a bacterial lacZ gene fused to only stripe 2 module of
even skipped regulatory element and injected back into
the fly embryo
Even skipped regulatory region
What does this finding
reveal?
Recombinant plasmid containing rat growth hormone fused to mouse metallothionein regulatory region in a bacterial plasmid vector
This recombinant plasmid,was injected into the mouse oocytes.
Promoter function seen in transgenic mice.
Mt-1 (metallothionein) gene is induced by heavy metal
A mouse derived from the eggs injected with growth hormone gene (left) and a normal littermate (right). The expression of a ‘transgene' mostly depends on the regulatory sequences which have been used in the design of the gene construct. A classical example is the transgenic giant mice in which the expression of a growth hormone (GH) gene is driven by the metallothionein promoter
Key conclusions: Genes regulate functions- GH Growth Promoters drive gene expression Promoter of each gene each gene is uniquely regulated
Which one is the mother?
top related