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Evolution of the mitochondrial proteome from the large-scale patterns to the nitty-gritty details

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Protistology
intact-looking to half-digested), but there are also
some naked chloroplasts with three surrounding
membranes in the ciliate cytoplasm. To identify
the origin of these chloroplasts, we attempted to
analyze PCR products obtained from isolated whole
ciliate cells using various specific primers. The
diatom-targeted primers yielded a probable nuclear
SSU rDNA, and phylogenetic analyses based
on Neighbor Joining showed that this sequence
pertains to the centric diatom genus Discostella
(Stephanodiscaceae). It was especially close to
that of the diatom symbiont of the dinoflagellate
Peridiniopsis penardii. We are now carrying out a
detailed study of the ciliate and its symbiont from
taxonomic and life history points of view.
EVOLUTION OF THE MITOCHONDRIAL
PROTEOME, FROM THE LARGE-SCALE
PATTERNS TO THE NITTY-GRITTY DETAILS
Huynen M.A., Duarte I., Szklarczyk R., Elurbe D.
CMBI, Radboud University Medical Centre, Nijmegen, Netherlands
huynen@cmbi.ru.nl
I will review what has been inferred about the changes at the level of the proteome that accompanied
the evolution of the mitochondrion from an
Alphaproteobacterium, quantifying the amount of
gene loss, gene replacement and gene gain. Most
of the loss and replacements that separate current
day mitochondria from Alphaproteobacteria took
place before the radiation of the eukaryotes. Detailed
analyses of the evolution of the mitochondrial
complexes like the ribosome and oxidative phosphorylation show that also the acquisition of new
proteins occurred mainly before the radiation of
the eukaryotes, supporting an early acquisition of
mitochondria in eukaryotic evolution. Secondly I
will analyze the accumulation of new supernumerary
subunits and assembly factors from pre-existing
protein families for one protein complex in detail:
complex I. There is a spectrum of protein function
conservation between the complex I representatives
and their non-complex I homologs. In general, the
new complex I proteins appear to have been recruited
from proteins that are active in mitochondria:
proteins one expects to be expressed when and
where complex I is active. Within complex I
and its assembly there are many cases of neofunctionalization after gene duplication, one case
of sub-functionalization, and one case in which a
complex I protein itself appears to have been the
source of the evolution of a new protein in another
complex. Complex I and its assembly can therewith
be regarded as a treasure trove for pathway evolution.
· 29
CONDITIONAL EXPRESSION SYSTEM IS
NOT SUITABLE FOR DEVELOPMENTAL
STUDIES IN LEISHMANIA
Ishemgulova A. 1, Kraeva N. 1, Faktorov á D. 2,
Podešvová L.1, Lukeš J.2,3,4, Yurchenko V.1,2,5
1
- Life Science Research Centre, Faculty of Science,
University of Ostrava, 710 00 Ostrava, Czech Republic
2
- Czech Academy of Sciences, 370 05 Č esk é
Budějovice (Budweis), Czech Republic
3
- Faculty of Sciences, University of South Bohemia,
370 05 České Budějovice (Budweis), Czech Republic
4
- Canadian Institute for Advanced Research,
Toronto, ON M5G 1Z8, Canada
5
- Department of Pathology, Albert Einstein College
of Medicine, Bronx, NY, 10461, USA
vyacheslav.yurchenko@osu.cz
The genus Leishmania unites parasitic protists of the
family Trypanosomatidae causing leishmaniases,
several closely related diseases that affect human
and animal populations mainly in the tropical and
subtropical regions. The clinical manifestations
vary from spontaneously healing skin lesions to
progressive and potentially fatal visceral infections.
Leishmaniases represent a global health problem
with over 350 million people at risk and an annual incidence rate of 2–10 million worldwide.
Conventional and conditional systems allow for a
controlled activation or repression of gene expression in time and space. Such systems are nowadays
widely used to analyse a variety of cellular processes
in numerous parasites including Leishmania.
A T7-driven, tetracycline-inducible system for
protein expression was established in a human
pathogen Leishmania mexicana. The gene expression in this strain is tightly regulated and dose- and
time dependent. We believe that it can be widely
used by the parasitology community to analyse
effects of genes of interest on biology, physiology
and virulence of parasites causing cutaneous
leishmaniases. This system was used to analyse gene
expression profiles during L. mexicana differentiation (procyclics, metacyclics, and amastigotes). The
transcription/translation of the gene of interest was
severely decreased upon Leishmania differentiation
into metacyclic and amastigotes. However, the
same expression profile was documented for the T7
polymerase. The expression was demonstrated to be
not locus-specific but dependent on untranslated
regions flanking open reading frames of studied
genes. We concluded that the previously established
conventional gene expression systems might have
certain limitations in their common applications.
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scala, large, patterns, evolution, proteome, nitty, detailsv, mitochondria, gritty
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