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Education is a central element of resettled families' lives, and providing support to parents and children to learn about and integrate into the education system is essential.

International audience; Seed storage is often accompanied by a progressive loss of germination vigour and viability. In the present study, we have used Arabidopsis thaliana (L.) Heynh. seeds as a model, and carried out differential proteomics to investigate seed vigour. In our system, based on a controlled deterioration treatment (CDT), we compared seed lots treated for different time periods up to 7 days. Germination tests showed a progressive decrease of seed vigour depending on the duration of CDT. Proteomic analyses revealed that loss in seed vigour can be accounted for by protein changes in the dry seed and by an inability of the low vigour seeds to display a normal proteome during germination. Furthermore, the CDT strongly increased the extent of protein oxidation (i.e. carbonylation), which will in turn induce a loss of functional properties of proteins and enzymes and/or enhance their susceptibility towards proteolysis. These results highlighted essential mechanisms for germinative quality such as translational capacity and mobilization of seed storage reserves.

International audience; The present report describes how the soluble germinal angiotensin 1-converting enzyme (gACE) appears in the epididymal fluid, where it has been identified in some laboratory rodents and domestic ungulates. We showed that this gACE results from an active proteolytic process that releases the enzyme's extracellular domain from sperm in a precise spatiotemporal location during epididymal transit and that this process involves serine protease activity. Using polyclonal antibodies against the C-terminal intracellular sequence of ACE, a fragment of approximately 10 kDa was detected on the sperm extract only in the epididymal region, where the gACE release occurs. The fluid enzyme was purified, and the cleavage site was determined by mass spectrometry to be between Arg(622) and Leu(623) of the mature sheep gACE sequence (equivalent to Arg(627) and Arg(1203) of the human mature gACE and somatic ACE sequences, respectively). Thereafter, the C-terminal Arg was removed, leaving Ala(621) as a C-terminal. Using an in vitro assay, gACE cleavage from sperm was strongly increased by the presence of epididymal fluid from the release zone, and this increase was inhibited specifically by the serine protease-inhibitor AEBSF but not by para-aminobenzamidine. None of the other inhibitors tested, such as metallo- or cystein-protease inhibitors, had a similar effect on release. It was also found that this process did not involve changes in gACE phosphorylation.

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National audience

chap. 18; International audience

In this study, we purified the first member of a new ribonuclease (RNase) A family from fluid of the proximal caput of the boar epididymis. This protein, named "Train A," is the most abundant compound secreted in the anterior part of the boar epididymis. After 2D electrophoresis, it is characterized by more than 10 isoforms ranging in size from 26 to 33 kDa and pI from 5 to 8.5. Several tryptic peptides were N-terminal sequenced, and an antiserum against one of these peptides was obtained. The protein was immunolocalized in the epididymal epithelium of the proximal caput, especially in the Golgi zone and the apical cytoplasm of the principal cells. In the lumen, spermatozoa were negative but droplets of reaction product were observed within the lumen. Full lengths of Train A cDNA were obtained from a lambdagt11 boar caput epididymis library and sequenced. The deduced protein is composed of 213 amino acids, including a 23-amino acid peptide signal and a potential N-glycosylation site. The mRNA of this protein has been retrieved and partially sequenced in the bull, horse, and ram, and homologous cDNA is found in databanks for the rat, mouse, and human. All the sequences are highly conserved between species. This protein and its mRNA are male-specific and exclusively expressed in the proximal caput of the epididymis, the only site where they have been found. Train A presents an RNase A family motif in its sequence. The RNase A family is a group of several short proteins (20-14 kDa) with greater and lesser degrees of ribonucleolytic activity and with supposed different roles in vivo. However, the presence of a long-conserved N-terminal specific sequence and the absence of RNase catalytic site for Train A indicate that Train A protein is a member of a new family of RNase A.

International audience