Slide 1

MITOCHONDRIAL DNA

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O BJEC TIVES:

 WHAT IS MITOCHONDRIAL DNA?

 WHAT ARE ITS CHARACTERISTICS?
 NUCLEAR DNA VS MITOCHONDRIAL DNA.
 MATERNAL INHERITANCE
 INTERACTIONS BETWEEN MITOCHONDRIAL AND
NUCLEAR DNA
 MUTATIONS IN MTDNA AND DISEASE
 MITOCHONDRIAL GENETIC BOTTLENECK

Slide 3 - Cellular respiration – ATP

production
Wh a t is Mito c h o n d ria ?
- Mitochondria are well-defined
 En e rg y Po w e r Pla n t o f th e C e ll
cytoplasmic organelles of the cell which
 O rg a n e lle –vita l fo r C e llu la r Re sp ira tio n take part in a variety of cellular
 Ma n y c o p ie s w ith in o n e c e ll metabolic functions. Survival of the cells
 O n ly o rg a n e lle w ith it’s o w n DNA requires energy to perform different
functions. The mitochondria are
important as the fact that these
organelles supply all the necessary
biological energy of the cell, and they
obtain this energy by oxidizing the
substrates of the Krebs cycle. Energy of
the cell is got from the enzymatic
oxidation of chemical compounds in the
mitochondria. Hence, the mitochondria
re referred to as the 'power houses' of
the cell.
Functions of mitochondria depends on the cell
type in which they are present.
The most important function of the
mitochondria is to produce energy. The simpler
molecules of nutrition are sent to the
mitochondria to be processed and to produce
charged molecules. These charged molecules
combine with oxygen and produce ATP
molecules. This process is known as oxidative
phosphorylation. 
Mitochondria help the cells to maintain proper
concentration of calcium ions within the
compartments of the cell. 
 The mitochondria also help in building certain
parts of blood and hormones like testosterone
and estrogen. 
The liver cells mitochondria have enzymes that
detoxify ammonia. 
The mitochondria also play important role in
the process of apoptosis or programmed cell
death. Abnormal death of cells due to the
dysfunction of mitochondria can affect the
function of organ. 
Slide 4
Wh a t is Mito c h o n d ria ?

Circ ula r m tDNA

Slide 5 Basically this not located in the nucleus.

Most cells contain at least 1000 mtDNA
MITO C HO NDRIAL DNA:
molecules distributed among hundreds of
individual mitochondria.
 This is consists of a circular chromosome,16.5kb in size that is located inside the
mitochondrial organelle.
 Not all the RNA and protein synthesized in a cell are encoded in the DNA of the
nucleus. Oxidative phosphorylation (or OXPHOS in



It contains 37 genes, and encodes 2 types of rRNA and 22 tRNAs
Genes encode 13 proteins that are subunits of enzymes of oxidative
short) is the metabolic pathway in

phosphorylation
The remaining 74 polypeptides of the oxidative phosphorylation complex are
which cells use enzymes to oxidize nutrients,
encoded by the nuclear genome.
thereby releasing energy which is used to
reform ATP. In most eukaryotes, this takes
place inside mitochondria.

Most of the energy requirement for cell growth,

differentiation, and development is met by the
mitochondria in the form of ATP produced by
the process of oxidative phosphorylation.
Human mitochondrial DNA encodes a total of
13 proteins, all of which are essential for
oxidative phosphorylation. The mRNAs for
these proteins are translated on mitochondrial
ribosomes. Recently, the genes for human
mitochondrial ribosomal proteins (MRPs) have
been identified. In this review, we summarize
their refined chromosomal location. It is well
known that mutations in the mitochondrial
translation system, i.e., ribosomal RNA and
transfer RNA cause various pathologies.

translation RNA

ribosomal RNA –translation

 13 genes used in cellular energy production

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NUCLEAR DNA VS MITOCHONDRIAL DNA

DNA
99%
100%

80%

60%

40% 1%
20%

0%

Slide 7
Nuclear DNA vs. Mitochondrial DNA

 Nuclear DNA  Mitochondrial DNA

 found in nucleus of the cell  found in mitochondria of the cell

 2 sets of 23 chromosomes  each mitochondria may have
several copies of the single
mtDNA molecule

 maternal and paternal  maternal only

 double helix  Circular
 bounded by a nuclear  free of a nuclear envelope
envelope
 DNA packed into chromatin
 DNA is not packed into chromatin

Slide 8 Plant mtDNA is far more complex than

animal mtDNA – larger size (300-600kb) and
C HARAC TERISTIC S
variable in size (up to 2000kb).
 Is inherited exclusively from the mother!
mtDNA is a circular shape single chromosome Introns and exons are parts of genes. Exons
It is only 16 kb in length
code for proteins, whereas introns do not. A


 Codes for 37 genes




Contains 22 tRNA and 2 rRNA coding genes
Encodes 13 proteins that are subunits of oxidative phosphorylation
great way to remember this is by considering
 It is approximately 16,569bp (base pairs) long. introns as intervening sequences and exons
Contains only exons, no introns
as expressed sequences.


 No repair mechanism-high mutation rate especially in D-loop!

 Replicative segregation, homoplasmy & heteroplasmy

According to researchers, there are an average

of 8.8 exons and 7.8 introns per human gene. (5)
mitochondrial DNA is not the same between all
populations and this is due to lack of repair
mechanisms and proofreading capabilities
which makes it susceptible to base substitutions,
leading to high mutation rates. The mutation
rates of mitochondrial DNA are 10 times higher
than in nuclear DNA.

Mitochondrial DNA (mtDNA) is known for

high mutation rates caused by lack of protective
histones, inefficient DNA repair systems, and
continuous exposure to mutagenic effects of
oxygen radicals.

Slide 9
Myoclonic epilepsy with ragged-red fibers
More than 100 different rearrangements and 100 different point

mutations have been identified in mtDNA that can cause human
disease,often involving the CNS and musculoskeletal system
 The diseases that result from these mutations show distinctive pattern
of inheritance because of 3 unusual features of mitochondria.
1.Replicative Segregation
2.Homoplasmy and Heteroplasmy
3.Maternal Inheritance
a displacement loop or D-loop is
a DNA structure where the two strands of a
double-stranded DNA molecule are separated
for a stretch and held apart by a third strand of
DNA.
Myoclonic epilepsy with ragged-red
fibers (MERRF) is a disorder that affects many
parts of the body, particularly the muscles and
nervous system. In most cases, the signs and
symptoms of this disorder appear during
childhood or adolescence. The features
of MERRF vary widely among affected
individuals, even among members of the same
family.
MERRF is inherited in a mitochondrial pattern,
which is also known as maternal inheritance.
This pattern of inheritance applies to genes
contained in mtDNA. Because egg cells, but not
sperm cells, contribute mitochondria to the
developing embryo, children can only inherit
disorders resulting from mtDNA mutations from
their mother. These disorders can appear in
every generation of a family and can affect both
males and females, but fathers do not pass traits
associated with changes in mtDNA to their
children.
In most cases, people with MERRF inherit an
altered mitochondrial gene from their mother,
who may or may not show symptoms of the
disorder. Less commonly, the disorder results
from a new mutation in a mitochondrial gene
and occurs in people with no family history
of MERRF.
MERRF is characterized by muscle twitches
(myoclonus), weakness (myopathy), and
progressive stiffness (spasticity). When the
muscle cells of affected individuals are stained
and viewed under a microscope, these cells
usually appear abnormal. These abnormal
muscle cells are called ragged-red fibers. Other
features of MERRF include recurrent seizures
(epilepsy), difficulty coordinating movements
(ataxia), a loss of sensation in the extremities
(peripheral neuropathy), and slow deterioration
of intellectual function (dementia). People with
this condition may also develop hearing loss or
optic atrophy, which is the degeneration
(atrophy) of nerve cells that carry visual
information from the eyes to the brain. Affected
individuals sometimes have short stature and a
form of heart disease known as cardiomyopathy.
Less commonly, people with MERRF develop
 fatty tumors, called lipomas, just under the
surface of the skin.

Mutations that cause MERRF impair the ability

of mitochondria to make proteins, use oxygen,
and produce energy. These mutations
particularly affect organs and tissues with high
energy requirements, such as the brain and
muscles. Researchers have not determined how
changes in mtDNA lead to the specific signs and
symptoms of MERRF.
Slide 10
Replicative segregation

 At cell division, multiple copies of mtDNA in each of

the mitochondria in a cell replicate and sort randomly
among newly synthesized mitochondria.
 The mitochondria in turn are distributed randomly
between the 2 daughter cells. This is called as
replicative segregation.
 The first unique feature of mitochondria is the
absence of tightly controlled segregation seen during
mitosis and meiosis of the 46 nuclear chromosomes.

Slide 11 A cell can have some mitochondria that have a

mutation in the mtDNA and some that do not.
Homoplasmy and Heterolasmy
This is termed heteroplasmy. The proportion of
mutant mtDNA molecules determines both
One daughter cell may by chance receive mitochondria that contain only a

pure population of normal mtDNA or a pure population of mutant the penetrance and severity of expression of
mtDNA(Homoplasmy)
 The daughter cell may receive a mixture of mitochondria some with and some diseases.
some without mutation(Heteroplasmy)

Homoplasmy refers to a cell that has a uniform

collection of mtDNA: either completely normal
mtDNA or completely mutant mtDNA.
A unique feature of mtDNA is that, at cell
division, the mtDNA replicates and sorts
randomly among mitochondria. In turn, the
mitochondria sort randomly among daughter
cells. Therefore, in cells where heteroplasmy is
present, each daughter cell may receive different
proportions of mitochondria carrying normal
and mutant mtDNA.

Homoplasmy is a term used in genetics to

describe a eukaryotic cell whose copies
of mitochondrial DNA are all identical.[1] When
in normal and healthy tissues, all cells are
homoplasmic.[2] Homoplasmic mitochondrial
DNA copies may be normal or mutated;
[1]
 however, most mutations
are heteroplasmic (only occurring in some
copies of mitochondrial DNA). It has been
discovered, though, that homoplasmic
mitochondrial DNA mutations may be found in
human tumors.
Slide 12
Slide 13
Maternal Inheritance of mtDNA
Sperm mitochondria are generally eliminated from the

embryo so that mtDNA is inherited from the mother.
 All children of a female who is homoplasmic for a
mtDNA mutation will inherit the mutation
 None of the offspring of a male carrying the same
mutation will inherit the defective DNA
Maternal inheritance of a homoplasmic mtDNA

mutation causing Leber Hereditary optic neuropathy is
known.
This means that all of the offspring of a female
will have identical and homoplasmic
mitochondrial DNA. It is very rare for females
to pass on heteroplasmic or homoplasmic
mutations because of the genetic bottleneck,
where only a few out of many mitochondria
actually are passed on to offspring.
LHON-  is the disease in humans that is most
frequently associated with homoplasmy.[7] This
condition is characterized by the atrophy
of retinal ganglion cells, which leads to central
blindness and eventually total blindness.
[11]
 Although it is passed down maternally, it is
seen more often in young men than in other
ages or sexes, which leads researchers to believe
that there are many other genetic or
environmental factors that contribute to
developing the disease.
Cancer
Some research has shown that an inherited
heteroplasmic mutation can cause cancer in
older age as cells become homoplasmic.[6] In
one study, doctors found that a cancer patient's
tumor consisted of only homoplasmic cells with
mutant mtDNA and that healthy cells in his
body were heteroplasmic for mutant
mtDNA. Additionally, researchers found that
the patient's siblings had the same
heteroplasmic mutation. This indicates that the
heteroplamic mutation was inherited, and over
time led to homoplasmic cells that caused
cancer.
Slide 14
Features of Maternal Inheritance

 1.Number of mtDNA molecules within the developing

oocyte is reduced before being amplified to the huge
total seen in mature oocytes.This restriction and
subsequent amplification of mtDNA during oogenesis is
termed Mitochondrial genetic Bottleneck.
 2.Variablity in the percentage of mutant mtDNA
molecules seen in the offspring of a mother with
heteroplasmy for a mtDNA mutation arises from the
sampling of only a subset of mtDNAs during oogenesis.

Slide 15 The mitochondrial genetic bottleneck.

Mitochondrial genetic Bottleneck During the production of primary oocytes, a
selected number of mitochondrial DNA
(mtDNA) molecules are transferred into each
oocyte. Oocyte maturation is associated with the
rapid replication of this mtDNA population.
This restriction-amplification event can lead to a
random shift of mtDNA mutational load
between generations and is responsible for the
variable levels of mutated mtDNA observed in
affected offspring from mothers with pathogenic
mtDNA mutations. Mitochondria that contain
mutated mtDNA are shown in red, those with
normal mtDNA are shown in green.

Slide 16
Mitochondrial Genetic Bottleneck

 All the children of a female who is homoplasmic for a

mtDNA mutation will inherit the mutation, whereas
none of the offspring of a male carrying the same
mutation will inherit the defective DNA.

Slide 17

 This is why often mtDNA is referred to as the “slave”

of the nuclear DNA, because mtDNA depends on many
nuclear genome-encoded proteins for its replication and
the maintenance of its integrity.
 Thus, diseases of oxidative phosphorylation arise not
only from mutations in the mitochondrial genome but also
from mutations in nuclear genes that encode oxidative
phosphorylation components.
 Mutations in many of these nuclear genes can lead to
disorders with phenotype similar to that of the mtDNA
diseases.
Slide 18
Why is mitochondrial mutation so high?

 The mitochondrial genome has a very high mutation rate, 10- to

17-fold higher than that observed in nuclear DNA. Although
mtDNA repair systems do exist and , they are not sufficient to
counteract the oxidative damage sustained by the mitochondrial
genome.
 Protective histones are also lacking.
 Oxygenation process is high percentage.
 The mtDNA mutation rate can be increased by environmental
agents or by mutation of nuclear genes involved in mtDNA
maintenance

Slide 19 mitochondrial disease which can cause muscle

Mutations in mtDNA and disease
weakness, diabetes, strokes, heart failure and
epilepsy.
 About 1 in 4,000 children in the US develop mitochondrial disease by the
age of 10 years
 mtDNA genome mutates 10 times more frequently than does nuclear DNA
 More than 100 different rearrangements and about 100 different point
mutations that are disease-causing have been identified in mtDNA.
 The clinical phenotype resulting from mtDNA mutations is diverse, however
the diseases of central-nervous or muscular-skeletal systems are most
common.
 Pleiotropy and variable expressivity is common in different affected family
members (due to heteroplasmy).
 Pleiotropy – multiple phenotipic effects of a single allele or pair of alleles.

Slide 20 mitochondrial disease which can cause muscle

Mutations in mtDNA and disease
weakness, diabetes, strokes, heart failure and
epilepsy.

Pleiotropy – multiple
phenotypic effects
of a single allele or
pair of alleles.
A pleiotropic gene is a single gene that controls
more than one trait.
occurs when one gene influences two or more
seemingly unrelated phenotypic traits.
Therefore, a mutation in a pleiotropic gene may
have an effect on several traits simultaneously
due to the gene coding for a product used by a
myriad of cells or different targets that have the
same signaling function.
An example of pleiotropy is phenylketonuria,
which is an inherited disorder that affects the
level of phenylalanine in the body.

Phenylalanine is an amino acid that can be

obtained from food. Phenylketonuria causes
this amino acid to increase in amount in the
body, which can be very dangerous. The human
disease is caused by a defect in a single gene on
chromosome 12 that affects multiple systems,
such as the nervous and integumentary system.
[2]
 Other examples of pleiotropy are albinism,
sickle cell anemia, and certain forms of autism
and schizophrenia. Pleiotropy not only affects
humans, but also animals, such as chickens and
laboratory house mice, where the laboratory
 house mice have found to exhibit the "mini-
muscle" allele.
Slide 21
THREE TYPES OF MUTATIONS

 It has been identified in mtDNA:

 (1) missense mutations in the coding regions of genes that
alter the activity of an oxidative phosphorylation protein;
 (2) point mutations in tRNA or rRNA genes that impair
mitochondrial protein synthesis;
 (3) Deletions or duplications of the mtDNA molecule. They
are generally somatic in origin, although a small proportion is
inherited, in some diseases.

Slide 22

 The number of mtDNA molecules within developing oocytes is reduced before

being subsequently amplified to the huge total seen in mature oocytes.
 This restriction and subsequent amplification of mtDNA during oogenesis is
termed the mitochondrial genetic bottleneck.
 Thus, mothers with a high proportion of mutant mtDNA molecules are more
likely to produce eggs with a higher proportion of mutant mtDNA and
therefore are more likely to have clinically affected offspring than are
mothers with a lower proportion.
 Also, for reasons unknown, deleted mtDNA molecules are generally not
transmitted from clinically affected mothers to their children.

Slide 23
Some diseases associated with mtDNA

 MERRF (Myoclonic Epilepsy with Ragged Red Fibres)

 MELAS (Myopathy, Epilepsy, Lactic acidosis, Stroke-
like episodes)
 LHON (Leber’s Hereditary Optic atrophy)
 Kearn-Sayre (eye problems, heart block, ataxia and
loss of coordination)
 Leigh syndrome (rare severe brain disease in infancy,
also heart problems)

Slide 24
References

 Ibrahim Okumus and Y. Çiftci / Turk. Turkish Journal of Fisheries and Aquatic Sciences 3: 51-
79 (2003)
 ARIAGNA LARA,* JOSELUIS PONCE DE, Molecular Ecology Resources (2010) 10,
421–430
 Vallone, P.M., Just, R.S., Coble, M.D., Butler, J.M., Parsons, T.J. (2004) A multiplex allele-
specific primer extension assay for forensically informative SNPs distributed throughout the
mitochondrial genome. Int. J. Legal Med., 118: 147-157. [Protocol for 11plex SNP assay
developed at NIST] [Genotyper macro for mtSNP 11plex]
 Coble, M.D., Just, R.S., O'Callaghan, J.E., Letmanyi, I.H., Peterson, C.T., Irwin, J.A.,
Parsons, T.J. (2004) Single nucleotide polymorphisms over the entire mtDNA genome that
increase the power of forensic testing in Caucasians. Int. J. Legal Med., 118: 137-146.
 Coble, M.D. (2004) The identification of single nucleotide polymorphisms in the entire
mitochondrial genome to increase the forensic discrimination of common HV1/HV2 types in
the Caucasian population. PhD dissertation, George Washington University, 206 pp.
 www.google.co.in/mtdna/wikipedia/in
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