LICAO, MARY LAVIENNE C.

MBIODX1 - LECTURE MIDTERMS

THE MOLECULAR NATURE OF GENES

FRIEDRICH MIESCHER
 From Tubingen, Germany
 1869 – isolated nuclei from pus cells in waste surgical
materials
 The nuclei contained novel phosphate-containing substance
(acidic) he called nuclein.
 NUCLEIN – is mostly chromatin (a complex of DNA and
chromosomal proteins).

BY THE END OF 19TH CENTURY

 DNA and RNA had been separated from the protein that
clings to them in the cell
 Both are suspended in proteins to make them more stable
but purification is made in order to separate them.
 This paved the way for a better analysis of the nature of
genetic material – DNA and RNA.

TRANSFORMATION IN BACTERIA DNA: THE TRANSFORMING MATERIAL

FREDERICK GRIFFITH OSWALD AVERY, COLIN MACLEOD AND MACLYN MCCARTY (1944)
 Laid the foundation for the identification of DNA as the  They made use of the transformation test similar to Griffith’s
genetic material. work.
 Thru this experiment on transformation in Streptococcus  They defined the transforming material from Griffith’s work by
pneumoniae. purifying the extract from S. pneumoniae

2 TYPES OF S. PNEUMONIAE ANALYTICAL TOOLS USED:

WILD TYPE MUTANT STRAIN ULTRACENTRIFUGATION  They spun the transforming
Spherical cells surrounded by a Spherical cells with no capsule material and the material with the
capsule transforming activity sedimented
Colonies are large, glistening Colonies are small and rough rapidly suggesting a very high
and smooth molecular weight – a
Virulent (encapsulated) – able Avirulent – it cannot cause a characteristics of DNA.
to cause a disease disease

ELECTROPHORESIS  The transforming material had a

relatively high mobility – also a
characteristic of DNA because of
its high charge or mass ratio.

GRIFFITH’S WORK
UV ABSORPTION  The transforming material’s
 The heat-killed virulent colonies of S. pneumoniae could
SPECTROPHOTOMETRY absorption spectrum matched
transform avirulent cells to virulent ones.
that of DNA.
 The virulent trait is passed from the dead cells to the live,
 The light it absorbed most strongly
avirulent one.
had a wavelength of about 260
 The virulent trait was even passed on to their descendants as
nm in contrast to proteins which
a heritable trait.
absorbs maximally at 280 nm.
 The gene for virulence was gained during transformation.
 The transforming substance in the heat-killed bacteria
was probably the gene for virulence itself.
 What is the transforming material?
 There is a substance in S. pneumoniae that causes it to ELEMENTARY CHEMICAL  Yielded an average
be virulent and that substance can be passed on even ANALYSIS nitrogen/phosphorus ratio of 1.67,
in a heritable form but they do not know what it is. about what one would expect for
DNA.
 DNA is rich in nitrogen and
phosphorus.
 Protein is rich in nitrogen but poor
in phosphorus.
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
JAMES WATSON AND FRANCIS CRICK (1953) 4. It is like the clinching experiment for us to find out the
 Published the double helical model of the DNA structure transforming material, the one that carries genetic
 Most widely accepted by most scientists information is the DNA

THE CHEMICAL NATURE OF POLYNUCLEOTIDES

NUCLEOSIDES
 Formed by joining a 5 carbon sugar and a nitrogenous base

PURINE NUCLEOSIDES PYRIMIDINE NUCLEOSIDES

the glycosidic linkage is the glycosidic linkage is
between the C1 of the sugar between the C1 of the sugar
and the N9 of the purine base and N1 of the purine base

ERWIN CHARGAFF
 Shown that the bases were not really found in equal
proportions in DNA (or merely a repetition of ACTG) THE NUCLEOSIDES
 The base composition of DNA varied from one species to BASE RNA NUCLEOSIDE DNA NUCLEOSIDE
another Adenine Adenosine Deoxyadenosine
 The amount of Adenine is almost always equal to Thymine Guanine Guanosine Deoxyguanosine
 The amount of Guanine is equal to Cytosine Cytosine Cytidine Deoxycytidine
Uracil Uridine Not visibly found
Thymine Not visibly found (deoxy)thymidine
 The difference between the two is not only the absence and
presence of Uracil and Thymine but also the sugar
 RNA – ribose sugar / Uracil
 DNA – deoxyribose sugar / Thymine

NUCLEOTIDES
 The basic structural unit of nucleic acids
 It is composed of:
1. 5 CARBON SUGARS
2. NITROGENOUS BASES
THE HERSHEY-CHASE EXPERIMENT 3. PHOSPHATE GROUP
 Made up of phage T2 (a bacteriophage that infects E. Coli)
 They labeled the protein with sulfur and to label the nucleic
acid which in this case is DNA phosphorous which shows
green coloration in this illustration:

5 CARBON RIBOSE
SUGARS  Found in RNA
 It has a hydroxyl group in the 2nd carbon
atom

1. Both proceeds in infecting the cell (Infection) DEOXYRIBOSE

2. And then they blended it using a waring blender (the  Found in DNA
common household blender); it was used because if they will  Do not have oxygen on the 2nd carbon
use centrifugation, there is a possibility that the cell will be atom
destroyed along the process. Whereas, if you’re going to use
waring blender, the phage will just be remove from being
attached in the surface of bacteria (blending)
3. The next step is centrifugation in determining which is found
inside the cell. What they are able to find out was there is
presence of phosphorous, it means that what enters the cell
is the genetic material, the nucleic acid or more specifically
the DNA.
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
NITROGENOUS PYRIMIDINE BASE  Cytosine
BASES  Uracil
 Thymine
PURINE BASES  Adenine
 Guanine

PHOSPHATE  Phosphoric acid ester of nucleosides

GROUP  Acidic in nature – nucleic acids
 FOR DEOXYRIBOSE  This structure shows a TRINUCLEOTIDES
 Phosphorylation occurs of C3 and  It is a string of three nucleotides
C5  It has polarity
 C1 and C4 – involved in fumanose  The top of the nucleotide bears the free 5’ phosphate group
ring (5’ end)
 C2 – no hydroxyl group  The bottom has a free 3’ hydroxyl group (3’- end)

 FOR RIBOSE
 Phosphorylation occurs at C2, C3 DNA STRUCTURE
and C5
 C1 and C4 – involved in fumanose
EXPERIMENTAL  Proposed by Creek and Watson
ring
BACKGROUND  LINUS PAULING
 Started it because of his study about
proteins and then he found out that
DIFFERENCE BETWEEN NUCLEOSIDE AND NUCLEOTIDES proteins also follow an Alpha-helix feature
NUCLEOSIDE NUCLEOTIDES  Alpha-helix feature of the protein
 composed of sugar and a  composed of phosphate structure
base group, sugar and base - The idea of the helix structure especially if
 Link that joins sugar to the  Link that joins nucleoside it’s a long structure it usually follows a helix
base in a nucleoside is to a phosphate is a shape
Beta- glycosidic linkage phosphordiester bond ERWIN  He was able to compute for percentages of
CHARGAFF adenine and guanine

CHARGAFF’S RULE
 The amount of adenine is almost always equal
to the amount of thymine (A=T)
 The amount of guanine always equal to the
amount of cytosine (G=C)
 The amount of purine bases will be equal the
amount of pyrimidine (PURINE=PYRIMIDINE)
 The amount of A+T is not equal to the amount of
G+C: this ration differs among organism but
same in different tissues of the same organism
 CHARGAFF’S RULE EXEMPTIONS: single stranded
viruses don’t follow this rule
 Explained base pairing regularities or
“complementary relationships” among
organisms
 Chargaff’s rule was out before the structure of
Creek and Watson
 It was one of the things that helped with
their double helix structure
 It is true for every organism

THE NUCLEOTIDES
1. Adenylic acid
2. Guanylic acid
3. Cytidylic acid
4. Uridylic acid
5. Thymidic acid
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
MAURICE WILKINS AND ROSALIND FRANKLIN THE DOUBLE HELIX
 Used x-ray attraction to analyze the three-dimensional  Likened to a twisted ladder
structure of DNA  The curving sides of the ladder = sugar phosphate backbone
PROCESS  The rungs of the ladder = base pairs
1. A very concentrated solution of DNA was prepared  The spacing between base pairs is 3.4 Å (Angstrom)
2. With a needle, a fiber was pulled out from the solution  The overall helix repeat distance is about 34 Å (Angstrom)
3. The fiber is a whole batch of DNA forced into side by side  There are about 10 base pairs per turn of the helix
alignment by the pulling action of the needle  The diameter of the DNA molecule is 20 Å
4. The fiber was enough like a crystal that it diffracted xrays in  The two strands are antiparallel
an interpretable way  If one has 5'-3' polarity from top to bottom, the other
must have 3' - 5" polarity from top to bottom

THE X-RAY RESULT

 A series of spots arranged in an Xshape indicates that the
DNA structure itself must be very simple.
 By contrast, a complex irregular molecule like a protein gives
a complex x-ray diffraction pattern with many spots, like a
surface peppered by a shotgun blast.
 Since DNA (composed of millions of nitrogenous bases) is very HYDROGEN BONDING
large, it can be simple only if it has a regular repeating
structure, and the simplest repeating shape that a long, thin
molecule can assume is a corkscrew or helix

Actual photo taken by Rosalind Franklin

T-A 2 Hydrogen bond
IMPORTANT INSIGHTS IN ROSALIND'S X-RAY RESULT
C-G 3 Hydrogen bond
 It gave important information about the size and shape of
the helix
 The spacing between the adjacent spots in the arm of the X is "It has not escaped our notice that the specific base pairing we
inversely related to the overall repeat distance in the helix have proposed immediately suggest a possible copying
(34 Angstroms) mechanism for the general material" -Crick and Watson
 The spacing from the top of the X to the bottom is inversely
related to the spacing between the repeated elements in the  Since one strand is a complement of the other, the two
helix strands can be separated
 Each strand can serve as atemplate for building a new
CRICK AND WATSON: THE DOUBLE HELIX partner
 DNA is a double helix with its sugar-phosphate backbone on
the outside and its bases on the inside
 Purines are paired with pyrimidine
 This way the double stranded DNA will be uniform,
composed of very similarly shaped base pairs,
regardless of the unpredictable sequence of either
DNA strand by itself.
 Adenine must always pair with thymine and guanine with
cytosine
 The major groove will be composed of around 10
nitrogenous bases bago siya mag turn
 Single stranded DNA has polarity (5 prime and 3 prime
end)
 Nitrogenous bases – responsible for forming the
hydrogen bonds
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
 DNA is the direction in both strands, signified by a 5’ and 3’
end.
 The 5’ and 3’ end signifies which side group is attached the
DNA backbone.
 The 5’ end has a phosphate (P) group attached, while
3’ end has a hydroxyl (OH) group attached.
 This directionality is important for replication as it only
progresses in the 5’ to 3’ direction.
 However, the replication for is bidirectional
 One strand is oriented in the 3’ to 5’ direction (leading
strand) while the other is oriented 5’ to 3’ (lagging
strand).
 The two sides are therefore replicated with two different
processes to accommodate the directional difference.

RNA PRIMER

A. Initial DNA structure

B. DNS structure opens up
C. Complementary base pairing happens (happens during S
phase of cell cycle)

DNA REPLICATION

REPLICATION ENZYMES
DNA HELICASE  unwinds and separates double
stranded DNA as it moves along the
H=Hati DNA.
 It forms the replication fork by
STEP 2: PRIMER BINDING
breaking hydrogen bonds between
nucleotide pairs in DNA.  The leading strand is the simplest to replicate
 Once the DNA strands have been separated, a short place
DNA PRIMASE  a type of RNA polymerase that of RNA called a primer binds to the 3’ end of the strand
generates RNA primers.  The primer always binds as the starting point for replication
 PRIMERS  DNA PRIMASE
- are short RNA molecules that act as – enzyme that gererates primers
templates for the starting point of
DNA replication.

DNA POLYMERASE  synthesize new DNA molecules by

adding nucleotides to loading and
lagging DNA strands.

TOPOISOMERASE OF  unwinds and rewinds DNA strands to

DNA GYRASE prevent the DNA from becoming
tangled or supercoiled

EXONUCLEASES  group of enzyme that removes

nucleotide bases from the end of a
DNA chain. STEP 3: ELONGATION
 DNA POLYMERASE
DNA LIGASE  joins DNA fragments together by – enzyme responsible in creating the new strand by a process
forming phosphodiester bonds called elongation
between nucleotides.
TYPES OF DNA POLYMERASE
STEP 1: REPLICATION FORK FORMATION (IN BACTERIA AND HUMAN CELLS)
 The double stranded DNA molecule is “unzipped” into two Polymerase I,II, IV  responsible for error checking
single strands. and V and repair
 In order to unwind DNA, the interactions between base pairs
must be broken. Polymerase III  the main replication enzyme in
 DNA HELICASE bacteria
 Disrupts the hydrogen bonding between base pairs to  It can also bind to the strand at
separate the strands into a Y shape knows as the the site of the strand during
replication fork. replication
 REPLICATION FORK
- is the area where the template for replication will Polymerases alpha,  primary polymerases involved I
begin. delta, and epsilon DNA replication is eukaryotic
cells
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
 Because replication proceeds in the 5’ and 3’ direction on 2. CONSERVATIVE  The parental molecule directs
the leading strand, the newly formed strand is continuous. MODEL synthesis of an entirely new
 The lagging strand begins replication by binding with multiple double-stranded molecule,
primers such that after one round of
 Each primer is only several bases apart replication, one molecule is
 DNA polymerase then adds pieces of DNA, called OKAZAKI conserved as two old strands
FRAGMENTS, to the strand between primers  This is repeated in the second
 This process of replication is discontinuous as the newly round
created fragments are disjointed.  Old strand = pink color (does
not change)
STEP 4: TERMINATION
 EXONUCLEASE
- removes all RNA primers from the original strands once both
the continuous and discontinuous strands are formed.
 The primers are replaced with appropriate bases
 Another exonuclease “proofreads” the newly formed DNA to 3. DISPERSIVE MODEL  Material in the two parental
check, remove and replace any errors strands is distributed more or
 DNA LIGASE joins Okazaki fragments together forming a less randomly between two
single unified strand. The ends of the linear DNA present a daughter molecules
problem as DNA polymerase can only add nucleotides in the  In the model shown below, old
5’ to 3’ direction material is distributed
symmetrically between the two
 TELOMERES daughters molecules
 found at the ends of the parent strands and consists of  Other distribution are possible
repeated DNA sequence  Daughter strands = may halo
 Act as protective caps at the end of the chromosomes (blue and pink)
to prevent nearby chromosomes from fusing.

 TELOMERASE
 A special type of DNA polymerase that catalyzes the
synthesis of telomere sequences at the ends of the DNA
 Once completed, the parent strand and its
complementary DNA strand coils into the familiar
double helix shape SEPARATING TWO STRANDS OF DNA
GC CONTENT  Varies from one DNA to another
In the end, replication produces two DNA molecules, each with (PERCENTAGE OF G +  The differences in the percentage of
one strand from the parent molecule and one new strand. C) G + C are reflected in differences in
the physical properties of DNA
 Have a significant effect on its Tm
MODES OF REPLICATION  DNA with more GC will have a higher
 MOST USED Tm
1. SEMI-CONSERVATIVE  The two parental strands  GC (Guanine and Cytosine)
MODEL separate and each makes a base pairs are held by 3
copy of itself hydrogen bonds
 After one round of replication,  AT (Adenine and Thymine) base
the two daughter molecules pairs are held by 2 hydrogen
each comprises one old and bond
one new strand  It will take more energy to
 Note that after two rounds, two destroy 3 hydrogen bonds
of the DNA molecules consist  If there is very high GC content in a
only of new material, while the particular DNA segment you will
other two contain one old and need a higher temperature in order
one new strand to separate
 Daughter strand = blue in the  The GC content of a natural DNA can
photo vary from less than 25% to 75%
 This has a strong effect on the
 The semi-conservative model is physical properties of the DNA i.e. Tm
the intuitively appealing model (melting temperature) and density
because separation of the two  Tm and density is directly
strands provides two templates, proportional to the GC content of the
each of which carries all the DNA
information of the original  For example, if you are
molecule. It also turns out to be denaturing a DNA and it
the correct one (Meselson & requires higher melting
Stahl 1958). temperature, it would mean
that the DNA that you are trying
to denature have higher GC
content
 As well as the density is high
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
DNA  Happens when the two strands of the DNA  The higher the concentration, the more
DENATURATION/ DNA come apart CONCENTRATION likely it is that the complementary
DNA MELTING  Happens when the DNA solution is strands will encounter each other
heated enough to weaken the non- within the given time
covalent forces that hold the two  The higher the concentration, the
strands together to break it faster the annealing
 Hydrogen bonding is the non-
covalent forces that hold the two RENATURATION  The longer the time allowed for
strands together TIME annealing, the more will occur
 We need to overcome this force to
separate the two strands Cot (concentration  The product of the initial DNA
and time) concentration (Co) in moles of
MELTING  The temperature at which the DNA nucleotides per liter and time (t) in
TEMPERATURE (TM) strands are half denatured seconds
 The amount of strand separation or  The extent of renaturation of
melting is measured by the complementary strands in a DNA
absorbance of the DNA solution at solution will depend on Cot.
260 nm
 Once the DNA has already Cot ½  The Cot at which strands are half
separated it can be subjected annealed
to UV spectrophotometer and  There is linear relationship between
set at 260 and we observe the DNA complexity and Cot ½
absorption. If it is already half  The more complex a DNA, the
denatured it will absorb at 260 higher its Cot ½
nm

 HYPERCHROMIC SHIFT
- happens when the two strands of
DNA separate and the absorbance
quenching disappears thereby
increasing the absorbance by 30% -
40%

ORGANIC SOLVENTS  Dimethyl sulfoxide

 Formamide

PH  high pH disrupts the hydrogen

bonding between DNA strands and
promote denaturation

SALT  Lowering the salt concentration aids

CONCENTRATION in denaturation by removing the ions
that shield the negative charges on
the two strands from one another
 At low ionic strength, the mutually
repulsive forces of these negative
charges are strong enough to
denature the DNA at a low
temperature

ANNEALING / RENATURATION
 The process of reuniting the separated DNA strands

FACTORS THAT AFFECT ANNEALING/RENATURATION

TEMPERATURE  <250C is the melting temperature of the
DNA
 Low enough not to promote
denaturation
 High enough to allow rapid dilution of
DNA molecules and to weaken the
transient bonding between
mismatched sequences and short
intra-strand base-paired regions
 Rapid cooling after denaturation will
frustrate renaturation

 QUENCHING
– plunging the hot DNA into ice to
keep the DNA denatured
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
GENE EXPRESSION STRUCTURE
 Best explained making use of the central dogma PRIMARY STRUCTURE  Results of peptide bonds
 The linear order of amino acids
 Amino acids are joined by peptide
CENTRAL DOGMA
bonds
 tells us that the DNA makes a copy of one messenger RNA
wherein it undergoes transcription to produce the messenger
RNA. The messenger RNA is then translated to a protein. There
is 1 gene and 1 protein theory.

SECONDARY STRUCTURE  Results from the interactions of the

amino acids with their neighbors
 Results of hydrogen bonding

Alpha  from hydrogen

Helix bonding with near
 This process, the DNA is double stranded
neighbor amino
 During transcription, you open up a certain area of the DNA acids
wherein a certain gene that is needed for a certain protein
will open up Beta  involves extended
 And then, a template is made from it. Making use of a pleated protein chains,
messenger RNA sheet packed side-byside,
 This messenger RNA is then translated into proteins and that is that interact by
how genes are expressed hydrogen binding

Turn  connects a-helix with

b-pleated sheet
elements in a protein

TERTIARY STRUCTURE  Allows three-dimensional shape of a

polypeptide
 Results from disulfide bonds

KINDS
Globular catalytic, hormones
Fibrous structural support

1 gene will make 1 polypeptide

* If one particular polypeptide is defective, if you produce a
QUATERNARY STRUCTURE  Two or more individual polypeptides fit
defective protein, it will lose its functionality. That is how important
together in a complex protein
translation and transcription.  Highest level of protein structure

PROTEIN STRUCTURE
AMINO ACIDS  Structural component of proteins
 There are 20 known amino acids
 There 4 nucleotides in DNA, but
they code in 20 amino acids
 The arrangement of amino acids with
their distinct side chains gives each
protein its unique character PROTEIN AND THEIR BIOCHEMICAL FUNCTIONS
 most especially the side chains; STRUCTURE  for animals it is structural proteins which are
chemically speaking, it makes the chief constituents of skin, bones, hair
them very reactive and fingernails
 Amino acids are joined together using COLLAGEN  found in bones
peptide bonds to form polypeptides  provides the support for
calcium to bind and
POLYPEPTIDES  a chain of amino acids form the solid structure
 It has polarity of the bones
(+) N terminus/amino terminus KERATIN  hair and nails
(-) carboxyl terminus/ c-terminus
CATALYSIS  virtually all the reactions that take place in
the living organisms are catalyzed by
proteins called enzymes.
 Without enzymes, the reactions would take
place so slowly as to be useless

TRYPSIN catalyzes hydrolysis of

sucrose
DEHYDROGENASE converts ethanol to
acetaldehyde
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
MOVEMENT  every time we crook our finger, climb stairs, MESSENGER RNA (mRNA)
AND or blink an eye, we use our muscles.  DNA resides in the nucleus of eukaryotic cells, whereas
CONTRACTION  MUSCLES EXPANSION AND CONTRACTION proteins are made in the cytoplasm. This means that
are involved in every movement we make. something must carry information from one place to another
 Muscles are made up of protein molecules  Carry the genetic information from the genes to the
called MYOSIN AND ACTIN. ribosomes, which synthesize the polypeptides
 Each gene in a DNA produces separate mRNA when certain
ELASTICITY ELASTIN protein is required by the cell
 Protein which possesses unique elasticity  A template made from DNA
and strength, enables the skin, ligament  It carries the code (coden) that directs the synthesis of
and blood vessels to stretch and rebound proteins
 The size of the mRNA depends of the number of nucleotides
TRANSPORT  a large number of proteins fall into this in that particular gene
category
Albumin transports bilirubin, calcium, THE GENETIC CODE
fatty acids, some drugs  Codons/Coding triplet: groups of 3 adjacent bases that specify
Ferritin and transport iron an amino acid
transferrin  64 codons
Transcortin cortisol (cortisol-binding  Most amino acids are coded for by more than one codon
protein)
Hemoglobin carries oxygen CODONS
Lipoproteins cholesterol and triglycerides UUU Phenylalanine
UUA Leticine
HORMONES Many hormones are proteins, among them:
 Insulin AUG Methionine (start codon)
 Oxytocin UAU Tyrosine
 Human growth hormone CAU Histidine
 Endomorphines UGG Tryptophan

PROTECTION  when a protein from an outside source or  AUG = start codon

other substance (called antigen) enters  UAA, UAG, UGA - stop/termination codons
the body, the body makes its own proteins  The first 2 bases are more significant and important
(called antibodies) to counteract the  The 3rd base is valuable and sensitive to mutation
foreign protein.
 This is the major mechanism the body used CHARACTERISTICS OF CODONS
to fight diseases. UNIVERSAL All plant & animals have the same codon
 BLOOD CLOTTING to specify each amino acid
 another protective device carried DEGENERATE CODE more than one codon can specify the
out by proteins called fibrinogen. same amino acid
 Without clotting we would bleed to
CONTINUOUS Code does NOT overlap; code is read
death from any small wound.
sequentially
STORAGE  some proteins are used to store materials,
in the way that starch and glycogen store
energy. INTRONS EXONS
Casein in store nutrients for newborn
milk, mammals and birds. Found in eukaryotes only Found in both prokaryotes and
Ovalbumin in eukaryotes
eggs
Ferritin protein in the liver that stores Non-coding areas of the DNA Coding areas of the DNA
iron
Non-coding part of hmiRNA, which Nucleotide sequence in mRNA,
REGULATION  some proteins control the expression of are removed before translation by which codes for proteins
genes are thereby regulate the kind of RNA splicing to form mRNA
proteins manufactured in a particular cell,
and control when such manufacture takes Sequence of the introns frequently Exons are highly conserved
place changes over time. In other words,
they are less conserved

PROTEIN ASTRUCTURE AND FUNCTION DNA bases found between exons DNA bases that are translated into
 One - gene - one - polypeptide hypothesis proteins
 Each polypeptide is usually encoded in one gene
 Many enzyme contain more than one polypeptide chain Removed in the nucleus before the Mature mRNA contains exons and
mRNA moves to the cytoplasm moves to the cytoplasm from the
 Phenotypes can be expressed by only one gene
nucleus
 Defective proteins are coded by a defect in the DNA
• Example: Alkaptonuria and sickle cell anemia  The presence of exons and introns allows for greater
molecular evolution through the process of exon shuffling.
LICAO, MARY LAVIENNE C. MBIODX1 - LECTURE MIDTERMS
INTRONS
 Helps in the regulation of transcription where it protects
mRNA that leads to protein synthesis
 Control some genes that are involved in the transcription.
 Helps in gene expression and gene regulation that is it helps
copy the information through generations, especially in
humans, which have more introns than exons,
 Variations in introns are used in DNA fingerprinting and DNA
profiling, in forensics, potential and immigration cases.
 Ued by evolutionary biologists to study the degree to which
organisms are related
 Spliceosomes remove introns during post transcriptional
processes

TRANSCRIPTION
RNA polymerase
 enzyme that directs transcription

3 PHASES OF TRANSCRIPTION
INITIATION  A POLYMERASE recognize and binds tightly
to the promoter region
 This causes localized melting, or separation
of the two DNA strands within the promoter
 Approximately 10 base pairs are melted

 The polymerase starts building the RNA

chain using the four ribonucleoside
triphosphates: ATP, GTP., CTP, UTP
 The initiating substrate is usually a purine
nucleotide
 This enzyme joins a second nucleotide to the
first forming the initial phosphodiester bond
 Several nucleotides may be joined
before the polymerase leaves the
promoter and elongation begins

ELONGATION  RNA polymerase directs growing RNA chain

in the 5' to 3' direction the sequential
binding of ribonucleotides 3’ end of the
strand
 This moves along the DNA template and the
bubble of melted DNA moves along with it
 Exposed nucleotides in the DNA bubble pair
with complementary bases: C to G and A to
U

TERMINATION  Use terminators - used to signal termination

 They work in conjunction with RNA
polymerase to loosen the association
between RNA product and DNA template
 Result: RNA dissociates from the RNA
polymerase and DNA
 Transcription is thereby terminated

DIFFERENCE BETWEEN REPLICATION AND TRANSCRIPTION

 RNA polymerase makes only one strand:
 transcription is said to be asymmetric and contrasts with
semi-conservative DNA replication (Both DNA strands
are replicated
 DNA melting is limited and transient:
 only enough strand separation occurs to allow the
polymerase to read the DNA template strand.
 In replication, two parental strands separate
permanently