GENOMAS DEL

CLOROPLASTO Y LA MITOCONDRIA

prof. Jorge Hernández T., Ph.D.

La Teoría Endosimbiótica

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
 Evidence for Endosymbiotic Theory
• Mitochondria have very similar characteristics to purple-aerobic bacteria. They both
use oxygen in the production of ATP, and they both do this by using the Kreb’s Cycle
and oxidative phosphorylation. Similarly, chloroplasts are very similar to
photosynthetic bacteria in that they both have similar chlorophyll that harnesses light
energy that is converted into chemical energy.

• Mitochondria and chloroplasts are similar in size to bacteria, 1 to 10 µm.

• Mitochondria and chloroplasts DNA (circular), RNA, ribosomes, chlorophyll (for

chloroplasts), and protein synthesis is similar to that for bacteria. It was also
determined that mitochondria and chloroplasts divide independently of the cell they
live in.

• Both mitochondria and chloroplasts have double phospholipid bilayers. This appears
to have arisen by mitochondria and chloroplasts entering eukaryotic cells via
endocytosis.

• Strong phylogenetic evidence

• No histones
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
Gram (-) Gram (-)

prof. Jorge Hernández T., Ph.D.

Transferencia horizontal
de genes

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
 Mechanisms of DNA transfer from chloroplast and
mitochondrial DNA to the nucleus

• Bulk DNA – recombination between escaped organelle DNA and

nuclear DNA
– Experimental DNA transfer in yeast
– Non-coding sequence frequently transferred
– Whole organelle sequences transferred
– Degradation of the organelle genomes in pollen could make
DNA fragments available for uptake
(MUST HAPPEN)
• cDNA intermediates
– Nuclear copies of organelle genes often lack organelle-specific
introns and edited sites
(MAY HAPPEN)

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
Reproducción in vivo de
la Transferencia

prof. Jorge Hernández T., Ph.D.

Nuclear Chloroplast
promoter promoter

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
 Chloroplast Gene Transfer

Rate estimates from tobacco chloroplasts

– 1 transfer in 5 million leaf cells
– 1 transfer in 16 000 pollen grains

Higher rates of transfer in the pollen?

Degradation of the organelle genomes in pollen could
make DNA fragments available for uptake

prof. Jorge Hernández T., Ph.D.

Transferencia y adaptación
al genoma nuclear

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
 Why are the organelle genomes
maintained?

• Hydrophobicity -hydrophobic proteins are

poorly imported (excess of membrane
embedded proteins)
• Fitness advantage if coding sequence and
regulation are in same location
• Other constraints (e.g. RNA editing,
genetic code)

prof. Jorge Hernández T., Ph.D.

Genoma cloroplásticos
secuenciados

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
El genoma cloroplástico

prof. Jorge Hernández T., Ph.D.

 Chloroplast DNA (cpDNA)
General features:
1. double-stranded, circular molecule
2. no histones, but have bound proteins (e.g.,
Hu), organized into nucleoids
3. G-C content typically less than nuclear DNA
4. multiple copies (~30-100) per plastid (i.e., all
cp genes are multi-copy)
5. can be 10-20% of the total DNA in leaves

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
Major protein complexes of the chloroplast thylakoid membrane
of Arabidopsis thaliana

Polypeptide subunits encoded in the chloroplast

Polypeptide subunits encoded in the nucleus

Allen etHernández
prof. Jorge al., 2011T., Ph.D.
prof. Jorge Hernández T., Ph.D.
 Volvox carteri

Dunaliella salina

Chlamydomonas reinhardtii

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
Genes cloroplásticos

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
Ribulose-1,5-bisphosphate carboxylase/oxygenase,
RuBPCase (Rubisco)
• Catalyzes carboxylation of ribulose-1,5
bisphosphate: CO2 + RuBP  3PGA (x 2)

• 2 subunits, large (LS) and small (SS)

– 8 copies of each per holoenzyme

• LS gene (rbcL) in the chloroplast

• SS gene (rbcS) in the nucleus

• extremely abundant, because inefficient

– Pyrenoid in algae is mostly RuBPCase

• regulated during light-dark cycles

– enzyme more active in the light
– also synthesized mainly in the light

prof. Jorge Hernández T., Ph.D.

 Translational regulation of RuBPCase LS by SS

Incoming SS somehow promotes translation of rbcL mRNA

Buchanan et al.
There also seems to be autoregulation of rbcL translation:
Cohen et al. (2006) Plant Physiol. 141, 1089; Wostrikoff and Stern (2007) PNAS 104, 6466
prof. Jorge Hernández T., Ph.D.
 Retrograde Signaling & Regulation

Retrograde Regulation
- Regulation of nuclear genes by
the chloroplast
- Nuclear genes typically
encode chloroplast proteins
- Signaled by:
(1) Developmental state of the
plastid/gene expression
(2) Photo-oxidative stress

Anterograde Regulation
- Regulation of chloroplast genes
by nuclear gene products
- Occurs at most levels of
expression

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
 Translational regulation of RuBPCase LS by SS

Incoming SS somehow promotes translation of rbcL mRNA!

Bogorad
lab

Fig. 9.16 in Buchanan et al.

There also seems to be autoregulation of rbcL translation:
Cohen et al. (2006) Plant Physiol. 141, 1089; Wostrikoff and Stern (2007) PNAS 104, 6466
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
The Protein Import Machinery of Chloroplast

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
El genoma mitocondrial

prof. Jorge Hernández T., Ph.D.

 Human Mitochondrial DNA (mtDNA)

The human mitochondrion contains 5-10 identical, circular

molecules of DNA. Each consists of 16,569 base pairs
carrying the information for 37 genes which encode:
• 2 different molecules of ribosomal RNA (rRNA)
• 22 different molecules of transfer RNA (tRNA) (at
least one for each amino acid)
•13 polypeptides
The rRNA and tRNA molecules are used in the machinery
that synthesizes the 13 polypeptides.
The 13 polypeptides are subunits of the protein complexes in
the inner mitochondrial membrane, including subunits of
NADH dehydrogenase, cytochrome c oxidase, and ATP
synthase. However, each of these protein complexes also
requires subunits that are encoded by nuclear genes,
synthesized in the cytosol, and imported from the cytosol into
the mitochondrion. prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
Complete mitochondrial
genome maps for
Dunaliella salina,
Chlamydomonas
reinhardtii
Volvox carteri

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
The Protein
Import
Machinery of
Mitochondria

prof. Jorge Hernández T., Ph.D.

El genoma cloroplástico vs
el mitocondrial

prof. Jorge Hernández T., Ph.D.

prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.
prof. Jorge Hernández T., Ph.D.