A kinase-anchoring protein (AKAP)95 recruits human chromosome-associated protein (hCAP)-D2/Eg7 for chromosome condensation in mitotic extract.

Association of the condensin multiprotein complex with chromatin is required for chromosome condensation at mitosis. What regulates condensin targeting to chromatin is largely unknown. We previously showed that the nuclear A kinase-anchoring protein, AKAP95, is implicated in chromosome condensation. We demonstrate here that AKAP95 acts as a targeting protein for human chromosome-associated protein (hCAP)-D2/Eg7, a component of the human condensin complex, to chromosomes. In HeLa cell mitotic extract, AKAP95 redistributes from the nuclear matrix to chromatin. When association of AKAP95 with chromatin is prevented, the chromatin does not condense. Condensation is rescued by a recombinant AKAP95 peptide containing the 306 COOH-terminal amino acids of AKAP95. Recombinant AKAP95 binds chromatin and elicits recruitment of Eg7 to chromosomes in a concentration-dependent manner. Amount of Eg7 recruited correlates with extent of chromosome condensation: resolution into distinct chromosomes is obtained only when near-endogenous levels of Eg7 are recruited. Eg7 and AKAP95 immunofluorescently colocalize to the central region of methanol-fixed metaphase chromosomes. GST pull-down data also suggest that AKAP95 recruits several condensin subunits. The results implicate AKAP95 as a receptor that assists condensin targeting to chromosomes.

pEg7, a new Xenopus protein required for mitotic chromosome condensation in egg extracts.

We have isolated a cDNA, Eg7, corresponding to a Xenopus maternal mRNA, which is polyadenylated in mature oocytes and deadenylated in early embryos. This maternal mRNA encodes a protein, pEg7, whose expression is strongly increased during oocyte maturation. The tissue and cell expression pattern of pEg7 indicates that this protein is only readily detected in cultured cells and germ cells. Immunolocalization in Xenopus cultured cells indicates that pEg7 concentrates onto chromosomes during mitosis. A similar localization of pEg7 is observed when sperm chromatin is allowed to form mitotic chromosomes in cytostatic factor-arrested egg extracts. Incubating these extracts with antibodies directed against two distinct parts of pEg7 provokes a strong inhibition of the condensation and resolution of mitotic chromosomes. Biochemical experiments show that pEg7 associates with Xenopus chromosome-associated polypeptides C and E, two components of the 13S condensin.

Effects of Oxyrrhis marina (Dinoflagellata), bacteria and vitamin D(2) on population dynamics of Tisbe holothuriae (Copepoda).

Laboratory experiments were performed to determine which food combinations had the most favorable effect on population dynamics of the harpacticoid copepod Tisbe holothuriae. In addition to basic diets of TetraMin (T), rice (R) or a mixture of both (T+R), supplements consisting of the following were given: the heterotrophic dinoflagellate Oxyrrhis marina, a bacterial suspension and vitamin D(2). Results show a beneficial effect of bacteria at a given concentration on T. holothuriae, which is enhanced in combination with vitamin D(2). A higher concentration of bacteria together with vitamin D(2), however, has a reverse effect leading to greater mortality, lower sex ratio and number of ovigerous females, and a dramatic decrease in production of eggs, regardless of the basic diet. We conclude that different concentrations of a given bacterial community may have variable effects on the reproduction of harpacticoid copepods and that, in addition to the bacteria, vitamin D(2) may also play a role in invertebrate nutrition.

The Xenopus laevis aurora-related protein kinase pEg2 associates with and phosphorylates the kinesin-related protein XlEg5.

We have previously reported on the cloning of XlEg5, a Xenopus laevis kinesin-related protein from the bimC family (Le Guellec, R., Paris, J., Couturier, A., Roghi, C., and Philippe, M. (1991) Mol. Cell. Biol. 11, 3395-3408) as well as pEg2, an Aurora-related serine/threonine kinase (Roghi, C., Giet, R., Uzbekov, R., Morin, N., Chartrain, I., Le Guellec, R., Couturier, A., Dorée, M., Philippe, M., and Prigent, C. (1998) J. Cell Sci. 111, 557-572). Inhibition of either XlEg5 or pEg2 activity during mitosis in Xenopus egg extract led to monopolar spindle formation. Here, we report that in Xenopus XL2 cells, pEg2 and XlEg5 are both confined to separated centrosomes in prophase, and then to the microtubule spindle poles. We also show that pEg2 co-immunoprecipitates with XlEg5 from egg extracts and XL2 cell lysates. Both proteins can directly interact in vitro, but also through the two-hybrid system. Furthermore immunoprecipitated pEg2 were found to remain active when bound to the beads and phosphorylate XlEg5 present in the precipitate. Two-dimensional mapping of XlEg5 tryptic peptides phosphorylated in vivo first confirmed that XlEg5 was phosphorylated by p34(cdc2) and next revealed that in vitro pEg2 kinase phosphorylated XlEg5 on the same stalk domain serine residue that was phosphorylated in metabolically labeled XL2 cells. The kinesin-related XlEg5 is to our knowledge the first in vivo substrate ever reported for an Aurora-related kinase.

[Development of bacterial populations of telluric, enteric, and marine origin in planktonic systems exposed to urban pollution; mathematical data analysis]. / Etude de l'évolution des populations bactériennes d'origine tellurique, entérique et marine dans des systèmes planctoniques soumis à une pollution urbaine, traitement mathématique des données.

A study of the comparative distribution of different types of bacterial populations in the sea area where the city of Marseille rejects its waste water shows that, when in contact with the marine environment, a great part of the allochthonic bacterial population undergoes physiological stress expressed by an abrupt reduction of the effectives and by weak values of the energetic charge. The presence of coliforms and streptococci in waste water lens, at important distances from discharging conduits, shows the possibility of a large diffusion for a fraction of the bacterial populations that withstand the initial stress. From mathematical analyses, an inverse correlation is revealed between detergents and the bacterial population as a whole, and between phenols and various allochthonic bacterial populations. The above results were evidenced through a sampling strategy based upon a 24-h-cycle study and a search for mathematical correlations between several biological, chemical, and physical parameters.