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Ongoing DNA transfer from the mitochondrial to the nuclear genome
Miria Ricchetti (mricch@ pasteur.fr)
Bioinformatic and Comparative Genome Analysis CourseHKU-Pasteur Research Center, Hong Kong, China
August 17-29, 2009
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1. The genomes of eukaryote cells
The genomes of eukaryote cells
chloroplasts (in plants)
mitochondria
nucleus
Endosymbiont theory
Mitochondria and chloroplasts are relics of free-living bacteria that formed a symbiotic association with the
precursor of the eukaryotic cell
Origin of mitochondria and chloroplasts
EUCARYOTES
Algae
Plants
Fungi
ProtozoaSlime moulds
Gram-positive bacteria
Spirochetes
Green photosynthetic
bacteria
EUBACTERIA
Methanogens
Extreme halophiles
Thermoacidophiles
Sulfolobus
Thermoplasma
ARCHAEBACTERIA
UNIVERSAL ANCESTOR
Animals
Purple photosynthetic
bacteria
Mitochondria
Cyanobacteria
Chloroplasts
Origin of mitochondria and chloroplasts
Mitochondria generate most of the cell's supply of adenosine triphosphate (ATP), used as a source of chemical energy.
Mitochondria are also involved in other processes, such as signaling, cellular differentiation, cell death, as well as the control of the cell cycle and cell growth.
Mitochondria have been implicated in several human diseases, including mitochondrial disorders and cardiac dysfunction, and may play a role in the aging process.
Mitochondria
McBride HM et al. 2006. Curr. Biol. 16: R551-600;Bossy-Wetzel E, et al 2003. Curr Opin Cell Biol 15, 706-16..
View of mitochondria within a cell
Chloroplasts, also called plastids, are organelles found in plant cells and other eukaryotic organisms that conduct photosynthesis. During photosynthesis, chloroplasts capture light energy to conserve free energy in the form of ATP and reduce NADP to NADPH.
Chloroplasts
Eberhard S. et al. 2008. Ann. Rev. Geneti. 42 463-515Puthiyaveetil S, Allen JF. 2009, Proc Biol Sci. 276:2133-45
85,779 bp
Mitochondrial genome
Saccharomyces cerevisiae
Nuclear genome
Homo sapiens 16,554 bp2.9 x 109 bp
1.2 x 107 bp
Genomes size
Reclinomonas americana
size mt genome (kbp)
protein-coding genes
67
Plasmodium falciparum 3
Homo sapiens 13
average 12-24
Saccharomyces cerevisiae 17
16.5
85.8
Smallest free-living -proteobacterial genomeBartonella henselae < 2000 1600
Arabidopsis thaliana 367
69
6
(protist)
and most vertebrates
(parasitic protist)
Mt genome size
45
Mitochondrial Genome Products
The mitochondrial genome:
• encodes some proteins but most proteins for organelle function are encoded by chromosomal genes and imported from cytoplasm
• encodes most of RNA components of protein synthesis (mRNA, rRNA, tRNA)
in Saccharomyces cerevisiae mt proteins are coded:
about 400 in the nucleus(200 are of bacterial origin)
17 in the mt
Genes coding for mitochondrialfunction and propagation
Mitochondrial genome products (in humans)
Protein Complex Encoded by Encoded by Mitochondrial NuclearGenome Genome
__________________________________________________________________
Oxidative phosphorylationNADH dehydrogenase 7 subunits >41 subunitsSuccinate CoQ reductase 0 subunits 4 subunitsCytochrome b-c1 complex 1 subunit 10 subunitsCytochrome c oxidase complex 3 subunits 10 subunitsATP synthase complex 2 subunits 14 subunits
Protein synthesis apparatus 2 rRNAs none 22 tRNAs none (13 mRNAs)
Ribosomal proteins none (~80 in total)
Other mitochondrial proteins none All, e.g., mitochondrial
DNA pol, RNA pol,other enzymes,
structural proteins
• Circular
• Much smaller than nuclear genome with a compact genetic organization
(human 16.5 kb, yeast 85 kb, melon 2500 kb)
• in humans:
• ~ 10 identical molecules per mitochondrion
(100s-10,000s mitochondria per cell)
• 37 genes ; no introns (but present in S. cerevisiae)
• 13 are protein-coding genes (respiratory complex)
• 24 are non-coding RNA genes
2 ribosomal RNAs, 22 transfer RNAs
Mitochondrial Genome
Genetic Organization of Human Mitochondrial DNA
ND1
ND2
CO1
CO2
CO3ND3
ND4L
ND4
ND5
CYB
16S rRNA
23S rRNA
ATPase 8
HSTRAND
LSTRAND
D-loop
H strand synthesis H strand transcription
L strand transcription
L strand synthesis
ND6
16.5 kb
DNSA
7
ATPase 6
Genes encoding proteins rRNA genes tRNA genes
Genetic organization of S. cerevisiae mitochondrial DNA
CO3
par
15S rRNA
CO1
ATPase 8ATPase 6
var
21S rRNA
CO2
85.7 kb
ATPase 9
Cyt
ochr
ome
b
Exons
Noncoding
Introns
Mitochondria and human diseases
Mitochondrial dysfunctions are responsible for several human diseases, in particular neurodegenerative disorders. This is the result of of the mitochondria's central role in energy production, reactive oxygen species (ROS) biology, and apoptosis. Mitochondrial DNA, whose integrity is strictly related to that of the mitochondrion itself, appears to be involved in pathologies and in the process of ageing. mtDNA mutations are sufficient by themselves to generate major clinical phenotypes
Bossy-Wetzel E et al. 2003. Curr Opin Cell Biol 15, 706-16.Scott SV, et al. 2003. Curr Opin Cell Biol 15, 482-8.Wallace DC & Fan W 2009 Genes Dev. 23:1714-36
Mitochondria and human diseases
Also defects in mtDNA maintenance are associated with an increasing number of human diseases (i.e. optic atrophy)
Spelbrink JN et al. 2001 Nat Genet, 28:223-31Trifunovic A et al. 2004 Nature, 429:417-23Alexander C et al. 2000 Nat Genet, 26:211-5.Delettre C et al. 2000 Nat Genet, 26:207-10
Mitochondria and human diseases
Some diseases are associated with the mutagenic insertion of mt DNA in the nuclear genome.
(it will be discussed later)
Turner C et al. 2003. Hum Genet 112, 303-9.Borensztajn K et al. 2002. Br J Haematol 117, 168-71Goldin E et al. 2004. Hum Mutat. 24, 460-5.Willett-Brozick JE et al. 2001. Hum Genet 109, 216-23.
protein-coding genes
average 130
ancestral cyanobacterial genome 3000
Chloroplast genome size
group of Apicomplexans 30
253Porphyra purpurea(red alga)
(parasitic protists)
• transfer to the nucleus
• function substituted by unrelated nuclear coded proteins (multi subunit RNA pol single subunit RNA pol)
• loss of function: ex. complex I (nad) in S. cerevisiae
Genome size reduction in mitochondria
• movement of nucleic acid from the mt to the nucleus• integration of the DNA in the nuclear genome• expression of the transfered gene (different codon usage)• protein must acquire a transit peptide to allow access to the organelle• coordination of the expression to different energy needs
Successful gene transfer requires:
from: Gray et al, Genome Biology (2001)
Phylogenetic trees constructed by nuclear and mt DNA, suggest thatmt and nuclear genomes have evolved in concert throughout much,if not most, of the evolutionary history of the domain Eukarya.
Why do organellar genes and up in the nucleus?
• haploid genome• relatively small size• maternal inheritance• normally does not undergo recombination• relatively rapid sequence evolution
higher risk of genetic drift
Characteristics of the mt genome
Some references and reviews on the endosymbiotic theory and on the origin of organelles
Margulis, Lynn, 1970, Origin of Eukaryotic Cells, Yale University Press.
Raven JA, Allen JF. 2003. Genomics and chloroplast evolution: What did cyanobacteria do for plants? Genome Biol. 4:209
Andersson SG, Karlberg O, Canback B, Kurland CG. 2003. On the origin of mitochondria: a genomics perspective. Philos. Trans. R. Soc. London Ser. B Biol. Sci. 358:165–7–9
Keeling PJ, Palmer JD. 2008. Horizontal gene transfer in eukaryotic evolution. Nat. Rev. Genet. 9:605–18
2. Intercompartemental DNA transfer
Theoretical DNA transfer in eukaryote cells
mitochondria
nucleus
chloroplasts (in plants)
Chloroplast DNA transfer in the nucleus
Chloroplast DNA transfer in the nucleus
• entire genes (during evolution)• infA in angiosperms• 33 kb of cpDNA on chromosome 10L of rice• transfer of chloroplast DNA into the nuclear genome of Nicotiana tabacum (preintegrated neomycine phosphotransferase gene) frequency: 1:16,000 • chloroplast DNA fragments (NUPT), ongoing
Millen et al, 2001 Plant Cell 13: 645; Yuan et al, 2002 Mol. Gen. Genet. 267: 713 Huang et al, 2003 Nature. 422: 472
Nuclear DNA transfer in mitochondria
Nuclear DNA transfer in mitochondria
• 1 possible case (intra or interspecies ?): gene homologue of bacterial MutS in the mt of the coral Sarcophyton glaucum
• interspecies (horizontal transfer): ribosomal (rps2 and rps11) and respiratory (atp1) proteins between distantly related flowering plants
Pont-Kingdon et al, 1998 J.Mol.Evol. 46: 419; Bergthorsson et al, 2003 Nature 424: 197
Chloroplast DNA transfer in mitochondria
Chloroplast DNA transfer in mitochondria
• tRNA set in higher plants
• 12 Kbp of DNA (originated from the inverted repeated region of the chloroplast genome) in maize • 5 independent transfers of the chloroplast rbcL to the mt genome in angiosperms (rbcL :ribulose-1,5-biposphate carboxylase/oxygenase)
• 17 stretches of plastid-like sequences found in the mt DNA of Oryza sativa L. (rice) 17-6653 bp; 6.3% of mt genome; 61-100% identity
Ellis J. 1982. Nature 299:678–79 ; Cummings et al, 2003 Curr. Genet 43: 131; Stern & Lonsdale, 1982 Nature 299: 698; Notsu et al, 2002 Mol. Genet. Genomics 268: 434)
Chloroplast DNA transfer in mitochondriaand then in the nucleus ?
In plants, a few (nuclear) genes that code for mt proteins are derived by duplication from nucleargenes of ancestrally chloroplast origin
(rps13 in some angiosperms, ribosomal protein S13 in Arabidopsis thaliana)
Adams et al, 2002 Plant Cell 14: 931; Mollier et al, 2002 Curr. Genet. 40: 405
Nuclear DNA transfer in chloroplasts
• two open reading frames, int and dpoB, in the large inverted repeat of the chloroplast genome of the green alga Oedogonium cardiacum no sequence similarity with genes in chloroplast genomesMt donor unkonwn
Nuclear DNA transfer in chloroplasts
Brouard JS et al. 2008. BMC Genomics 9:290
Mitochondrial DNA transfer in chloroplasts
not found… until now
Mitochondrial DNA transfer in the nucleus
Mitochondrial DNA transfer in nucleus
• entire genes (during evolution)• mt DNA frangments (NUMT), ongoing
• genes (ceased in animals, detected in plants)
Mt DNA insertions in the nuclear genome
ex. rsp10 gene (protein of the mt ribosome) in angiosperms
from: Knoop et al. 1995, Curr. Genet; Adams et al. 2000, Nature
Ongoing gene transfer from mt to the nucleus(in plants)
see: Sandoval et al. 2004 Gene 324:139
Ongoing gene transfer from mt to the nucleus (in plants)
SDH2
RPS14
RPL5 RPS14
Nucleus
Mitochondria rice
RPL5 RPS14 wheat
RPL5 RPS14 maize
RPL5 rice
RPL5 wheat
RPL5 maize
RPS14
RPS14
Bittner-Eddy et al. 1994 J. Mol. Biol. 235: 881
Dual expression (mt and nucleus) of atp9 in Neurospora
The nuclear and the mt copy of atp9 are expressedat different stages of the life cycle
(germinating spores versus vegetative cells)
Adams et al. 1999 PNAS 24:13863
Dual expression (mt and nucleus) of cox2 in legumes
• genes (ceased in animals, detected in plants)
• 620 kbp insertion in Arabidopsis thaliana
Mt insertions in the nuclear genome
from Stupar et al. 2001, PNAS 98, 5099. See also Lin et al. 1999, Nature 402, 761.
Mt DNA insertion (620 kbp)centromere
telomere
D D D BA AA C
A
D
B
C
duplications
Large mt DNA insertion in the centromeric regionof chromosome 2 of Arabidopsis thaliana
• genes (ceased in animals, detected in plants)
• 620 kbp insertion in Arabidopsis thaliana
• 7.9 kbp in felines
Mt insertions in the nuclear genome
A 7.9 Kbp insertion of mt origin in thenuclear genome of domestic cat
7.9 kb of a typically 17.0-kb mitochondrial genome inserted to a specific nuclear chromosomal position in the domestic cat.
the intergrated segment has subsequently become amplified38-76 times and now occurs as a tandem repeat macrosatellite
Lopez et al. 1994, J. Mol. Evol. 39, 554
Mt insertions in the nuclear genome
• genes (ceased in animals, detected in plants)
• 620 kbp insertion in Arabidopsis thaliana
• 7.9 kbp in felines
• DNA fragments of about 300 bp (NUMTs) till > 14 kbp (in most studied eukaryotes)
size
mt DNA (bp) nuclear (Mbps) NUMTs (BLASTN threshold 10-4
) % 10-3
Homo sapiens 16.571 2910 279.17 9,.6
Mus musculus 16.299 2500 53.453 2.1Rattus norvegicus 16.3 2800 6.34 0.2
Fugu rubripes 16.447 320 5.624 1.8Caernorhabdit is elegans 13.794 97 126 0.1
Drosophila melanogaster 19.517 122.7 534 0.4Anopheles gambiae 15.363 278.2 0 0.0
Ciona intestinalis 14.788 116.7 11.51 9.9Plasmodium falc iparum 5.697 22.9 152 0.7
Saccharomyces cerevisiae 85.779 12.5 1.241 9.9schisaccharomyces pombe 19.431 12.5 1.614 12.9
Arabidopsis thaliana 366.924 115.4 198.105 171.7Oryza sativa 490.52 420 409.104 97.4
Mitochondrial, nuclear and NUMT sizes in some eukaryotic genomes
from Richly & Leister 2004 Mol Biol Evol 21, 1081
Yeast species S. cerevisiae C. glabrata K. thermotolerans K. lactis D. hansenii Y. lipolytica
Nuclear genome size (Mb) 12.1 12.3 10.4 10.6 12.2 20.5
Mitochondrial genome size (kb) 85.7 20 23.5 40.2 29.4 47.9
NUMT number 32 14 1 8 145 47
Total transferred mitochondrial DNA (bp) 2356 1423 25 403 9377 2005
Transferred mitochondrial DNA (%) 2.7 7.1 0.1 1 31.8 4.2
NUMTs in six hemiascomycetous yeast species
from Sacerdot et al. 2008 FEMS Yeast 8, 846-850
Why variable abundance of NUMTs in different species ?
from Richly & Leister 2004 Mol Biol Evol 21, 1081
• frequency of DNA transfer ?• vulnerability of mt to stress & other factors?• n° of mt/cell ? (i.e. Plasmodium)• n° of somatic cell divisions from zygotes to meiosis• efficiency of nuclear import of mt DNA and/or its integrattion into the nuclear genome
• rate of loss of NUMTs ?• rates of DNA loss varies from fragments and among species• however, no NUMT loss has been show till now
Types of NUMTs and NUPTs
from Leister. 2006 Trends in Genetics 21, 655-663
3. How do NUMTs integrate in the nuclear genome?
Mitochondrial DNA integrate in the nuclear genome during double-strand break (DSB) repair in yeast
Ricchetti et al.1999 Nature, 402:96-100Yu & Gabriel 1999 Mol. Cell 4: 873-881
DSB
intact chromosome
mt DNA+
Ricchetti et al.1999, cited
Mt DNA insertion in the nuclear genomevia Non-Homologous End-Joining (NHEJ)
mt DNA insertionNHEJ
Analysis of mt insertions during the repair of DSBs in yeast
DSB
(A)
ATTACCCTGTTAT attattattttttattattaataataataatttatagggtttattctgttttatcataaatacgtaaatatctaacttagctctcaaattatattacT A A CAGGGTAAT
(ATAA)
ATTACCCTGTT A T ctttattatatttaagaatattattataattattattattattattatttttaataattaaaaatattaataataagtaaatattaattattgttcatttaatcattccaaaaatttaggtaatgatactgcttcgatcttaattggcatatttgcatgacctgtcccacacaactcagaacatgctc c ggccacgggagccggaaccccgaaaggaggaataagataaatatatagCAGGGTAAT
(T)
(A)
ATTACCCTGTTA agtttccatagaagtaataataataataaatatattaaatattaatata a t tattaattaaaataactaatttagatcaatctaaaaaatctaagtgtttagatgataataaagaatatttattaaagtattctattactttaatattttT A ACAGGGTAAT
ATTACCCTGT T A T ttagaatatttttaattaaataatataattaaatgaataccaaacttatattatatttatatttatatttatatttcT AACAGGGTAAT
(A)
34pAT9
34pAS16
34pAS15
622pBS8
ATTACCCTGTTAT3'TAATGGGAC 3'TATTGTCCCATTA
+ CAGGGTAATDigested I-SceI sites:
Mt insertions found at DSB sites in yeast
Ricchetti et al.1999, cited
NUMTs in the yeast nuclear genome
85,779 bp
Mitochondrial genome
Saccharomyces cerevisiae
Nuclear genome
1.2 x 107 bp
by DSB repair:
6 NUMTs size: single 47-97 bpmultiple 166-382 bp
homology :100%
by BLAST search:
30 NUMTs size: 22-230 bp homology :86-100%
Ricchetti et al.1999, cited
Yeast mitochondrial genome and origin of NUMTs
at DSBsin the yeast genomefound > once
NUMTS
4. Impact of NUMTs insertion on the eukaryotic genomes
• Sequence analysis of the genome of Homo sapiens
• PCR sampling of humans from different ethnic backgrounds
211 NUMTs detected in humans
Ricchetti et al.2004 PLoS Biology, 2:96-1313
NUMT
NUMT
amplified DNA fragment
NUMT present
NUMT absent
PCR amplification of NUMTsin the human genome
Insertion polymorphism of three NUMTs
1 2 3 4 5 6 7 8 9individual sex NUMT 1-74 NUMT 2-132 NUMT 2-53 NUMT 12-89 NUMT 13-75 NUMT 18-192 NUMT 11-541Caucasian-1 Female -/- +/+ -/- -/+ +/+ -/- +/+Caucasian-2 Male -/- +/+ +/+ -/- +/+ -/- -/-Caucasian-3 Male -/+ +/+ -/+ -/- +/+ -/- +/+Caucasian-4 Male -/- +/+ -/+ -/- +/+ +/+ * +/+Caucasian-5 Female -/+ +/+ -/+ -/- +/+ -/- -/+Caucasian-6 Male -/- +/+ -/+ -/- +/+ -/- -/+Caucasian-7 Female -/- +/+ -/- -/- +/+ -/- * -/+Caucasian-8 Male -/- +/+ +/+ -/+ +/+ -/- -/-Caucasian-9 Male -/- +/+ -/+ -/+ +/+ +/+ * +/+Caucasian-10 Female -/+ +/+ -/+ +/+ +/+ -/+ -/+
African-1 Male +/+ +/+ -/+ * -/- +/+ +/+ * -/-African-2 Female -/+ -/+ */* -/- -/- +/+ -/+ -/+African-3 Male -/- +/+ +/+ -/+ +/+ -/+ -/+
Pygmy-1 (Biaka) Male +/+ * +/+ -/+ -/- +/+ -/- -/-Pygmy-2 (Mbuti) Male -/- +/+ +/+ -/+ +/+ -/- -/-Pygmy-3 (Mbuti) Female -/- +/+ -/- -/+ +/+ -/+ -/-Pygmy-4 (Biaka) Female -/- +/+ -/+ * -/- +/+ -/- -/-
Chinese-1 Male -/+ +/+ -/- -/- +/+ -/+ -/-Chinese-2 Male -/- +/+ -/+ -/+ -/+ -/- * -/+
Japanese-1 Male -/- +/+ -/+ -/- -/+ -/+ -/+Japanese-2 Female -/- +/+ -/+ -/- +/+ -/- -/+
Insertion polymorphism of NUMTs in humans
Ricchetti et al.2004, cited
Frequency of alleles carrying the NUMT
NUMT %
2-132 9813-75 952-53 4818-192 2912-89 211-74 21
1 2 3 4 5 6 7 8 9individual sex NUMT 1-74 NUMT 2-132 NUMT 2-53 NUMT 12-89 NUMT 13-75 NUMT 18-192 NUMT 11-541Caucasian-1 Female -/- +/+ -/- -/+ +/+ -/- +/+Caucasian-2 Male -/- +/+ +/+ -/- +/+ -/- -/-Caucasian-3 Male -/+ +/+ -/+ -/- +/+ -/- +/+Caucasian-4 Male -/- +/+ -/+ -/- +/+ +/+ * +/+Caucasian-5 Female -/+ +/+ -/+ -/- +/+ -/- -/+Caucasian-6 Male -/- +/+ -/+ -/- +/+ -/- -/+Caucasian-7 Female -/- +/+ -/- -/- +/+ -/- * -/+Caucasian-8 Male -/- +/+ +/+ -/+ +/+ -/- -/-Caucasian-9 Male -/- +/+ -/+ -/+ +/+ +/+ * +/+Caucasian-10 Female -/+ +/+ -/+ +/+ +/+ -/+ -/+
African-1 Male +/+ +/+ -/+ * -/- +/+ +/+ * -/-African-2 Female -/+ -/+ */* -/- -/- +/+ -/+ -/+African-3 Male -/- +/+ +/+ -/+ +/+ -/+ -/+
Pygmy-1 (Biaka) Male +/+ * +/+ -/+ -/- +/+ -/- -/-Pygmy-2 (Mbuti) Male -/- +/+ +/+ -/+ +/+ -/- -/-Pygmy-3 (Mbuti) Female -/- +/+ -/- -/+ +/+ -/+ -/-Pygmy-4 (Biaka) Female -/- +/+ -/+ * -/- +/+ -/- -/-
Chinese-1 Male -/+ +/+ -/- -/- +/+ -/+ -/-Chinese-2 Male -/- +/+ -/+ -/+ -/+ -/- * -/+
Japanese-1 Male -/- +/+ -/+ -/- -/+ -/+ -/+Japanese-2 Female -/- +/+ -/+ -/- +/+ -/- -/+
Insertion polymorphism of NUMTs in humans
Ricchetti et al.2004, cited
NUMTs as genetic markers
Podnar et al 2007 J Mol Evol 64: 308-320. Unusual origin of a nulcear pseudogene in the Italian wallizard: intergenomic and interspecific transfer of a large section of the mitochondrial genome in the genus Podarcis (Lacertidae).
Vartanian JP, Wain-Hobson S. 2002 Proc Natl Acad Sci U S A 99: 7566-7569. Analysis of a library of macaque nuclear mitochondrial sequences confirms macaque origin of divergent sequences from old oral polio vaccine samples.
NUMTS as genetic tools to follow the geographic distribution of species or populations and to resolve phylogenetic ambiguities
Human-specific NUMTs
Specie-specific NUMTs
Ricchetti et al.2004 PLoS Biology, 2:96-1313Hazkani-Covo & Graur 2007 Mol. Biol. Evol. 24: 13-18
1 2 3 4 5 6 7 8 9 10 11 12NUMT code chr size (bp) % mt coord PCR size PCR size PCR amplification Chimp
identity no NUMT NUMT +/+ -/- +/- sequenced1-74 * 1 74 100 8919-8992 344 418 2 14 5 1 -1-90 * 1 90 94 8447-8542 927 1017 21 0 0 1 +
1-192 1 192 89 16386-16552 825 1017 21 0 0 2 +2-53 * 2 53 100 1762-1814 330 383 4 5 12 2 -
2-132 * 2 132 99 608-739 365 497 20 0 1 1 -
2-272 * 2 272 94 6735-7006 712a 984a21 0 0 0 -
3-76 * 3 76 96 4349-4424 389 465 21 0 0 1 +3-1323 3 1323 95 1392-2714 487 1810 20 0 0 0 -
4-93 * 4 93 96 14966-15058 326 419 21 0 0 0 -4-131 * 4 131 96 958-1088 300 431 21 0 0 0 -4-152 4 152 94 2895-3046 286 438 21 0 0 2 +4-240 4 240 91 2221-2460 868 1108 21 0 0 0 -
4-342 * 4 342 94 9323-9664 319a 661a21 0 0 0 -
5-949 5 949 79 6591-7902 919 21c0 0 1 +
5-8781 upd5 8781 88 6388-15168 507b 21c
0 0 3 +
5-8781 dwd5 8781 88 6388-15168 751b 21c
0 0 2 +6-527 6 527 90 2414-2930 709 1236 21 0 0 0 +7-100 * 7 100 98 1609-1704 338 438 21 0 0 1 -7-106 7 106 94 12946-13051 406 512 21 0 0 0 -7-164 7 164 96 2414-2570 270 434 21 0 0 1 +
7-505 * 7 505 85 1708-2212 260 765 21e0 0 0 +
8-59 * 8 59 94 803-861 638 697 21 0 0 0 +8-84 * 8 84 98 14846-14929 472 556 21 0 0 0 -
8-1470 * 8 1470 96 8405-8538 406b 21c0 0 2 +
11-72 * 11 72 98 14645-14716 244 316 21 0 0 0 -11-163 11 163 96 6636-6798 351 514 21 0 0 0 -
11-2451 11 2451 93 518-2968 400 2851 21 0 0 0 +12-68 * 12 68 94 4236-4303 351 419 21 0 0 0 +12-89 * 12 89 98 3786-3874 478 567 1 13 7 0 -13-75 * 13 75 94 9508-9578 260 331 19 0 2 0 -13-123 13 123 91 5103-5225 325 448 21 0 0 0 -13-256 13 256 99 978-1233 346 602 21 0 0 0 -
14-1023 14 1023 93 5578-6600 242 1265 0 21 0 0 -17-69 * 17 69 97 10128-10195 192 261 21 0 0 0 -17-653 17 653 95 6812-7464 452 1105 21 0 0 0 -18-156 18 156 94 14366-14521 374 530 21 0 0 1 -18-192 18 192 97 7969-8160 204 396 3 12 6 5 -20-70 * 20 70 92 12947-13016 445 515 21 0 0 0 -22-47 * 22 47 100 6176-6222 338 385 21 0 0 2 -X-267 X 267 95 684-950 293 560 21 0 0 0 -
Y-66 * Y 66 100 6494-6559 186 252 0 NDf 13f1 -
Y-71 * Y 71 100 1268-1338 361 432 0 8g 13g2 -
Y-3107 Y 3107 83 1155-4863 467 0 NDf 13f0 +
11-541 h 11 541 94 16074-60 195 736 4 8 8 0 ND
NUMT
A CB
PCR amplification and sequence analysis of NUMTs from humans and chimpanzees
Ricchetti et al.2004,cited
0
5
10
15
20
25
30
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X Y
total n° of NUMTs
human-specific NUMTs
50
100
150
200
250
n°
of N
UM
Ts
chro
mos
ome
size
(in
Mb
p)
Human-specific NUMTs in human chromosomes
No strict correlation between chromosome size and nb of NUMTs
Ricchetti et al.2004, cited
1 2 6 753 4 8 9 10 11
13
12
14 1615 17 18 19 20 21 22 X Y
Distribution of human-specific NUMTs in chromosomes
Ricchetti et al.2004, cited
5000 Alu sequences have colonised human chromosomesin the last 4-6 Myrs
from: Batzer & Deininger et al, 2002 Nature. Genet. Rev. 3, 370
180 Alu seq : 1 NUMT
27 NUMTs are specific to humans and have colonised humanchromosomes in the last 4-6 Myrs
Ricchetti et al 2004, cited
Colonisation rate of the human genome
NUMTs as potential mutagens
0
5
10
15
20
25
number of NUMTs
NUMTs common tohumans and chimpanzees
human-specific NUMTs NUMTs with insertionpolymorphism in humans
exon
intron
promoter
intergene
Insertion sites of NUMTs in the human genome
Ricchetti et al.2004, cited
80% NUMTs inserted in genesgenes (with introns) < 25% of the human genome
NUMTs preferentially insert in genes (in humans)
Diseases associated with the insertion of NUMTs
Diseases associated with the insertion of a NUMT include disorders as diverse as:
• a sporadic case of the Pallister-Hall syndrome, a multiple congenital anomaly characterised by hypothalamic hamartoma and bone deformities
Turner C et al. 2003. Hum Genet 112, 303-9
Diseases associated with the insertion of NUMTs
Diseases associated with the insertion of a NUMT include disorders as diverse as:
• a familial plasma factor deficiency, a severe type I factor VII deficiency resulting in severe bleeding
Borensztajn K et al. 2002. Br J Haematol 117, 168-71
Diseases associated with the insertion of NUMTs
Diseases associated with the insertion of a NUMT include disorders as diverse as:
• a mucolipidosis Type IV, showing a moderate phenotype of this usually severe neurodegenerative disorder
Goldin E et al. 2004. Hum Mutat. 24, 460-5
Diseases associated with the insertion of NUMTs
Diseases associated with the insertion of a NUMT include disorders as diverse as:
• a familial bipolar affective disorder associated to a constitutional chromosomal translocation with a NUMT at the junction site
Willett-Brozick JE et al. 2001 Hum Genet 109, 216-23
Diseases associated with the insertion of NUMTs
The diversity of these pathologies reflects a mutagenic process that can target a large variety of genes.
Three diseases related to NUMT insertions
NUMT
251 bp NUMT insertion inIVS acceptor splice site
Disease
severe type I factor (F) VII deficiency(rare bleeding disorder, family history)Borensztajn et al, 2001 Brit. J. Haemat. 117, 168
sporadic case of Pallister-Hall syndrome(de novo insertion)Turner et al, 2003 Hum. Genet. 112, 303
72 bp NUMT insertion into exon 14 of the GLI3 gene
41 bp NUMT insertion at thebreakpoint junction of a reciprocal
constitutional translocationt(9;11)(p24;q23)
bipolar affective disorder(cosegregation in family pedigree)
Willet-Brozick et al, 2001 Hum. Genet 109, 216
ctgcccagcctgctcagcctcacgcccgcccagcagtaccgcctcaaggccaag
ctgcccagcctgctcagcctcacgcccggtctaacaacatggctttctcaacttttaaaggataacagctatccattggtcttaggccccaaaaattttcccagcagtaccgcctcaaggccaag
LPSLLSLTPAQQYRLKAKLPSLLSLTPVSNNMAFSTFKG*
from: Turner et al, 2003 Hum. Genet. 112, 303
wt
+NUMT
wt+NUMT
Disease related to NUMT insertion:sporadic case of Pallister-Hall syndrome (GLI3)
17 known disease-related Alu insertions1,500,000 Alu seq/ human genome
4 known disease-related NUMT insertions211 NUMTs /human genome
Impact of new NUMT and Alu insertions on human diseases
NUMT insertions can modify the exon/intron pattern
hypothetical protein
1 32 4
1 32
a
bNUMT 12-89
hypothetical protein
1 2 3 4 5
1 2 3 4 65
a
bNUMT 17-653
hypothetical protein
1 2 3 4 5 6 7 8
1 2 3 4 5 6 7 8 9 10
a
bNUMT 5-8781
protein Q8N7L5
1 2 3 4 5 6 7
1 2 3 4 5 6 7 8
a
bNUMT 1-74
Some NUMT insertions in genes
Ricchetti et al.2004, cited
Generation of novel nuclear exons by NUMTs
Nuclear insertions of organelle DNA in yeast, H. sapiens, Arabidopsis, and rice:45 insertions contributed sequences to a total of 49 protein-coding exons in 34 genes.
Noutsos et al. 2007 Trends Genet. 23:597–601
NUMTs as regulatory sequences?
Discussion on unpublished data by Chatre and Ricchetti, 2009
• lysis of mitochondria ?• capture of cytoplasmic nucleic acids ? • illegittimate transport ?• membrane fusion ?
5. How do nucleic acids move to the nucleus?
Mitochondria-nuclear interactions
Human HeLa cells. Subpopulation of perinuclear mitochondria under normal growth conditions. By Laurent Chatre, Institut Pasteur
Perspectives
Analysis of the possible regulatory function of NUMTs in other eukaryotes,including humans
Analysis of mt-nuclear interactions and mt DNA release under normalgrowth and stress conditions
Acknowledgements
Laurent ChatreBenjamin Montagne
Cecile FairheadFredj TekaiaBernard Dujon
Unité de Génétique Moléculaire des Levures
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