Horloges de la Vie

Colloque Nature et Artifice , Fondation Singer-Polignac, 29/04/2014

Lhomme face a levolution de sa propre essence

Miroslav Radman

HORLOGE UNIVERSEL DE LVOLUTION (LIGNE GERMINALE) EST CONSTANT ET
CHRONOLOGIQUE:
La vitesse de lvolution des squences gnomiques est tonnamment
uniforme de la bactrie lhomme: 10
-9 /
paire des bases/an (M. Kimura, A.
Wilson).

HORLOGE SOMATIQUE (LA LONGVIT INDIVIDUELLE) EST BIOLOGIQUE,
SPECIFIQUE DE LESPCE, MAIS FLXIBLE:
La vitesse de dhorloge somatique (longvit) varie de 10.000 fois entre les
espces animales et elle augmente exponentiellement avec lge : la cintique
de la mortalit et morbidit est exponentielle (La loi de Gompertz).

QUELLE EST LA CHIMIE DE CES DEUX HORLOGES DA LA VIE?
Lhorloge de la vie et lhorloge du cancer sont corrls

La mort nest pas indispensable
La maladie nest pas invitable
Il existent des organismes qui resistent la mort et aux maladies: espces
immortelles (ci-dessous + Rotifres Bdelloides + rat taupe: une longevit
denviron 30 ans; le cancer jamais observ)


The marine medusa Turritopsis
Nutricula (pictured) and several
species (Hydra) reproduce and
essentially live forever
Many bacteria are effectively
immortal. This one (Deinococcus
radiodurans) is also incredibly
tough resisting massive radiation
doses
This strange beast (a Tardigrade)
can survive in space two weeks
in a vacuum at -272C exposed
to massive doses of UV-C and
cosmic rays
Human
LA LONGEVIT EST FLXIBLE (le nmatode):
Comme tout phnotype, la survie dpende du
gnotype et environnement, et ?
Garsin, Science Juin 2003
Longevit est flexible mme chez lhomme:
Age-related death rates for France 1979-2008
Evolution with age of all causes of death in France in 2008 normalized by the total population
for each age group: a common cause for all incurable age-related diseases ???
(compilation: Franois-Xavier Pellay)

About 90 % causes of death are age-related: living is the most life-threatening activity.
The exponential increase in colon cancer incidence with age
(5
th
power) typical of all major causes of human mortality.
Irreversible protein carbonylation accumulates exponentially in aging people
Age-related changes in the amount of oxidized proteins in cultured human dermal fibroblasts.
Is protein carbonylation just a biomarker of aging, its consequence, or its cause?
Normal individuals
Patients with progeria
Patients with Werner's syndrome

Data from:
Oliver et al., JBC 262:
5488-5491 (1987)
PROTEINS ARE NOT EQUALLY SUSCEPTIBLE TO OXIDATION
Senescent human cells accumulate oxidized proteins but
most of our proteins are inox (B. Friguet et al)
Nystrom 2005
Steady-state of oxydative damage and breakdown determines the level
of cellular proteome damage: the level of carbonylated proteins
reflects, at any time, the result of the rates of damage and the
selective protein turnover.
INTRACELLULAR PROTEIN CARBONYLATION (PC) DURING AGING OF DIVERSE SPECIES
PC IS BIOMARKER OF AGING IT MESURES BIOLOGICAL AGE; BUT, IS IT ITS CAUSE?
Data from Oliver et al 1987, Garland et al 1990, Adachi et al 1987, Starke-Reed et al 1989, Sohal et al 1993; our lab.

Effect of protein oxidation (red points) on a schematized cell
function
Protein Oxidation Correlates with High Mutation Rates
measured as MutL-eGFP foci (Elez et al., Curr Biol 2010)
WT
WY GroES/EL +++
WT del tig
mutH
mutH del tig
mutH GroES/EL+++
Krisko & Radman, PLoS Gen 2013
Spontaneous and radiation-induced protein carbonylation
in E. coli and Deinococcus radiodurans
Anita Krisko et M.R, PNAS, 2010
Radiation-induced Protein Carbonylation, not DNA damage,
Correlates with Cell Killing
Is protein carbonylation the
Chemistry of Death ?

YES!
(Krisko & Radman, PLoS Genetics, 2013)
A. Krisko & M. Radman, PNAS, 2010
Exogenous and endogenous sources of free radicals
PROTEIN
DAMAGE
Immortal stem cell status correlates with low protein cabonylation:
iPS-ing fibroblasts lowers their protein carbonylation,
i.e., protein damage is reversible!
Cell line nmol carbonyl
/ mg protein
Mouse Skin fibroblasts 9.26
P83 MEF P2 4.58
P83 iPS (10 pd)
CHO fibroblasts
2.43
3.88
28 ES cells I
P83 iPS (10
th
passage)
0.51
2.43
E. coli
D. radiodurans
2.O5
0.23
Anita Krisko, Alfonso Bellacosa & M.R. (unpublished)

NT
HCC
GRP90
HSP60
MutA
Preliminary analysis of a human Hepatocarcinoma (HCC) vs. Non-tumoral (NT) adjacent
tissue
(2D OxyDIGE): green is protein, red is carbonylation
Fernando Martin, unpubl.
Oxidation of three morphs of a human protein and the loss of their
function with age in the three carriers of these protein morphs
Split
Croatia

Chaperone activities lower proteome oxidation;
Low m.w. proteins are resistant to oxidation irrespective of chaperones
Martin Baraibar (B. Friguet lab) & Anita Krisko (unpubl)
Hill plot of radiation-induced protein carbonylation
in E. coli and D. radiodurans
ED50 Dra/Eco = 21
ED50 Dra/Eco = 25
PATHWAYS OF PROTEIN CARBONYLATION
Les causes de loxydation
The Causes of Protein Carbonylation
The Causes of Protein Carbonylation
Dukan et al., PNAS 2000
ROS up Susceptibility to ROS
up
La longevit est flexible: Augmentation continue de
lsprence de vie chez les femmes
UN estimate 1980
UN estimate 1990
UN estimate 2000
Oeppen & Vaupel Science 2002
Declining early/mid-life mortality Declining later-life mortality
Human Survival Curves 1860 vs. 1990

GRADIENT OF CELL MORBIDITY, DOWN TO MORTALITY, SOLELY BY INCREASING OXIDATIVE PROTEOME
DAMAGE: Manipulating protein synthesis fidelity (ribosomes) and folding accuracy (chaperones and
chaperonins) creates correlation between cell fitness and protein oxidation at constant levels of ROS!
ANTI-MUTATOR EFFECT AND CARBONYLATION SUPPRESSION BY 1mM
TROLOX (VIT E) SHOW THAT OXIDATION DURING MISFOLDING IS THE
ULTIMATE FUNCTIONAL (MUTAGENIC) DAMAGE.
Carbonyls
Mutation rate
+ Trolox
Krisko & Radman,
PLoS Genetics, 2013
Protein misfolding leads to an increased susceptibility to protein carbonylation:
Effect of single amino acid substitution on susceptibility to oxidative damage
Human a synuclein: A53T mutation in familial Parkinson's disease emerging at 30-35.
Radman-Krisko (prov.patent.appl.)