Mitochondria

Biogenesis
• Mitochondrial biogenesis can be defined as the growth and division of pre-existing
mitochondria.
• Mitochondrial biogenesis is the cellular process that produces new mitochondria. It’s
one of the ways cells adapt to the ever-changing energy requirements dictated by
environmental and physiological conditions.
• Mitochondria cannot be produced de novo; new mitochondria are produced by adding
new content—proteins, membranes—to pre-existing mitochondria.
• Mitochondria are direct descendants of an α-protobacteria endosymbiont that became
established in a host cell. Owing to their bacterial origin, mitochondria have their own
genome and can self replicate.
• Mitochondrial proteins are encoded by the nuclear and the mitochondrial genomes.
• The mtDNA (mitochondrial DNA) is a double-stranded circular molecule of approx. 16.5
kb that contains 37 genes encoding 13 subunits of the electron transport chain
complexes Complex I (NADH dehydrogenase), Complex III (Cytochrome bc1), Complex
IV (Cytochrome c oxidase), Complex IV (ATP synthase).
• The subunits ETC Complex II ( Succinate dehydrogenase) are encoded by nuclear
genome.
• The other mitochondrial gene products are the 22 tRNAs (transfer RNAs)
and two rRNAs (ribosomal RNAs) necessary for translation of the
respiratory subunit mRNAs within the mitochondrial matrix.
• Correct mitochondrial biogenesis requires the co-ordinated synthesis and
import of approx. 1000–1500 proteins encoded by the nuclear genome and
synthesized on cytosolic ribosomes .
• Mitochondrial biogenesis is a complex process that requires the replication
of mt-DNA, transcription and translation of mtDNA and nuclear DNA genes,
mitochondrial membrane recruitment and extension, protein import,
protein allocation to different mitochondrial compartments, and assembly
of the electron transport chain complexes in the mitochondrial membrane.
• Mitochondrial biogenesis increases metabolic enzymes for glycolysis,
oxidative phosphorylation and ultimately a greater mitochondrial metabolic
capacity.
• All this is carried out in coordination with the dynamics of the existing
mitochondrial network.
Import of Proteins
• Mitochondria are produced from the transcription and translation of genes
both in the nuclear genome and in the mitochondrial.
• The majority of mitochondrial protein comes from the nuclear genome, while
the mitochondrial genome encodes parts of the electron transport chain along
with mitochondrial rRNA and tRNA.
• As the majority of proteins are encoded in the nucleus, a mechanism for
targeting, import and correct assembly exists to ensure correct mitochondrial
function and shape.
• mRNAs are translated in the cytosol after activation of the nuclear genome to
precursor proteins.
• These precursor proteins are synthesized with an N-terminal positively charged
presequence capable of forming a basic, amphipathic α-helix.
• Precursor proteins will be transported to one of four areas of the
mitochondria, which include the outer membrane, inner membrane,
intermembrane space, and matrix.
• They traverse inner and outer mitochondrial membranes in an
unfolded conformation by crossing through protein translocases.
• The mitochondrial membrane potential and the action of matrix
Hsp70 (heat-shock protein 70) are used to drive translocation.
• The presequence is then cleaved by a matrix protease and, often with
the aid of molecular chaperones, the imported protein is folded.
• However, a majority of mitochondrial protein precursors use different
protein-import pathways to the other mitochondrial compartments.
• Precursor proteins that follow these routes do not typically contain
the N-terminal targeting signals, but harbour targeting information
within their mature sequence.
• Currently, four major membrane protein translocase complexes are
known.
• The TOM (translocase of the outer membrane) is the universal entry gate
for all proteins that are imported into mitochondria. The different protein
pathways then diverge at this point.
• TIM (translocase of the inner membrane) sorts matrix-targeted precursors.
• The PAM (presequence translocase-associated motor) regulates matrix
Hsp70 action to drive precursors into the matrix.
• Finally, the outer membrane SAM (sorting and assembly machinery) inserts
β-barrel proteins into the outer membrane. All of these processes are an
integral part of mitochondrial biogenesis.
• As polypeptides as delivered to their residence sites in the different
mitochondrial compartments, many associate with partner polypeptides
and with prosthetic groups to produce complexes.
 Fission and Fusion
• Mitochondria form a dynamic network that can change shape and size. These changes,
known as mitochondrial dynamics, are driven by two complementary processes:
mitochondrial fission and fusion.
• Mitochondrial fusion and fission are both controlled by GTPases of the dynamin family.
• Through mitochondrial fission, smaller mitochondria are created to which new
mitochondrial content can be added in mitochondrial biogenesis.
• The process of mitochondrial fission is directed by Drp1 (dynamin related protein), a
member of the cytosolic dynamin family. This protein forms a spiral around
mitochondria and constricts to break apart both the outer and inner membranes of
the organelle.
• A second protein important for mitochondrial fission is human fission protein 1
homologue (Fis1). This protein binds to the mitochondrial outer membrane with the
help of C-terminal trans-membrane domain.
• An increase in mitochondrial fission would create many fragmented mitochondria,
which has been shown to be useful for eliminating damaged mitochondria and for
creating smaller mitochondria for efficient transporting to energy-demanding areas.
• Mitochondrial fusion can be divided into two processes, the fusion of the mitochondrial
outer membrane followed by that of the inner membrane.
• The outer membrane fusion requires proteins known as mitofusins, (Mfn1 and Mfn2).
• Inner membrane fusion mainly involves an inner membrane-localized protein Opa1
(named based on its identification as a mutated gene in optic atrophy).
• The mitofusins are transmembrane dynamin-related GTPases, which induce the joining
of 2 mitochondrial “fragments” by forming dimers across the interface. The tethering of
mitochondria together is followed by GTP hydrolysis, which induces conformation
changes to cause mitochondrial fusion.
• Opa1 is a dynamin related GTPase that interacts with cardiolipin, a mitochondrial inner
membrane lipid, and is mostly found in cristae, consistent with its role in maintaining
cristae morphology.
• The upgraded mitochondrion can then fuse back with the existing mitochondrial
network, to create a larger and more elongated mitochondrion, that will be more
efficient in the generation of cell energy.
• Multiple research studies have observed correlated increases between mitochondrial
respiratory capacity with Mfn1, Mnf2, and Drp1 gene expression after endurance
exercises.
 Regulation of Biogenesis
• Multiple intrinsic cellular signalling pathways monitor mitochondrial
functioning and trigger organelle biogenesis.
• PGC-1 α , a member of the peroxisome proliferator-activated receptor gamma
(PGC) family of transcription coactivators, is the master regulator of
mitochondrial biogenesis.
• It is a co-transcriptional regulation factor that induces mitochondrial
biogenesis by activating different transcription factors like nuclear respiratory
factor 1 ( NRF-1) and nuclear respiratory factor 2 (NRF-2) which promote the
expression of TFAM (transcription factor A mitochondrial; also known as
mtTFA).
• NRF-1 controls the expression of a significant number of the proteins that
make up the five respiratory complexes, as well as proteins integral to
mitochondrial import and heme biosynthesis.
• It is also able to integrate nuclear control of the transcriptional and
replicative activity of the mitochondrial genome through the direct
modulation of transcription factor A mitochondrial (TFAM) and transcription
factor B proteins (TFBs) gene expression. NRF-2 is also able to regulate the
expression of proteins in the electron transport chain.
• NRF-1 and NRF-2 are important contributors to the sequence of events
leading to the increase in transcription of key mitochondrial enzymes, and
they have been shown to interact with TFAM, which drives transcription
and replication of mtDNA .
• PGC-1α stimulates mitochondrial biogenesis and respiration in muscle cells
through an induction of UCP (uncoupling protein) 2 and through induction
of NRF-1 and NRF-2 gene expression.
• Moreover, PGC-1α binds to and co-activates the transcriptional function of
NRF-1 on the promoter for Tfam
• PGC-1α also interacts with and co-activates other transcription factors such
as PPARs, thyroid hormone, glucocorticoid, oestrogen and ERRs (oestrogen-
related receptors) α and γ .
• ERRs are orphan nuclear receptors that target vast gene networks involved
in all aspects of energy homoeostasis, including fat and glucose
metabolism, as well as mitochondrial biogenesis and function .
• AMPK (AMP-activated protein kinase) is another major regulator of
mitochondrial biogenesis which regulates intracellular energy metabolism in
response to acute energy crises.
• It regulates mitochondrial biogenesis by phosphorylating and activating PGC-
1α upon sensing an energy deficiency in muscle.
• AMP kinase pathway monitors ATP levels, the calcium/calmodulin-
dependent kinase and p38 mitogen-activated kinase which monitors Ca++
homeostasis, mTOR a moderator of overall cell homeostasis, the cAMP/PKA
pathway, ROS signalling and NO and CO2 levels.
• These stimuli activate the synthesis of mitochondrial genes, both on mt-DNA
and nuclear DNA through a series of transcription factors mostly under
control of a family of coactivators (PGC-1α and PGC-1β).
• Signalling through PGC-1α is sufficient to increase total mitochondrial mass,
reactive oxygen species scavenging enzymes, oxidative phosphorylation
components, mitochondrial metabolic pathways, protein import complexes,
proteins involved in fission and fusion, and the levels of mitochondrial
sirtuins (protein regulating age and death of cells).
TORC [transducer of regulated CREB
(cAMP-response-element-binding
protein)-binding protein]
p38 MAPK (mitogen-activated protein
kinase)
SIRT1 [sirtuin (silent mating type
information regulation 2 homologue) 1]
PPARs- Peroxisome proliferator
activated receptor
CREB- cAMP Response Element Binding
Protein
YY1- Transcription factor
• The result of mitochondrial biogenesis is, effectively, an increase in
mitochondrial number and mass.
• Mitochondrial biogenesis is influenced by environmental stress such
as exercise, caloric restriction, low temperature, oxidative stress, cell
division and renewal and differentiation.