Canonical and Alternative Pathways in Cyclin-Dependent Kinase 1/Cyclin B Inactivation upon M-Phase Exit in Xenopus laevis Cell-Free Extracts

Cyclin-Dependent Kinase 1 (CDK1) is the major M-phase kinase known also as the M-phase Promoting Factor or MPF. Studies performed during the last decade have shown many details of how CDK1 is regulated and also how it regulates the cell cycle progression. Xenopus laevis cell-free extracts were widely used to elucidate the details and to obtain a global view of the role of CDK1 in M-phase control. CDK1 inactivation upon M-phase exit is a primordial process leading to the M-phase/interphase transition during the cell cycle. Here we discuss two closely related aspects of CDK1 regulation in Xenopus laevis cell-free extracts: firstly, how CDK1 becomes inactivated and secondly, how other actors, like kinases and phosphatases network and/or specific inhibitors, cooperate with CDK1 inactivation to assure timely exit from the M-phase

Mould walls inclination and dendritic morphology of CMSX-4 blades airfoils

The airfoils of single-crystalline turbine blades were studied. The blades made of industrial CMSX-4 superalloy were obtained by the Bridgman technique. Five different withdrawal rates (1–5 mm/min) were used. Series of as-cast samples were prepared by cutting the airfoils at different distances from the blade root. The metallographic sections, prepared for each cutting planes, were subjected to scanning electron microscopy observations and Laue diffraction studies. Macrostructure images of whole area of airfoil cross-sections were obtained by stitching several obtained SEM images. Morphology of dendrites in different area of cross-sections were analysed. The crystal orientation of each sample were determined by analysis of Laue pattern. It was found that the morphology of dendrites changes in the area, where the dendrite growth is limited by inclined mould walls

Translationally Controlled Tumor-Associated Protein

EditorialInternational audienceTranslationally-controlled tumor-associated protein (TCTP) has been discovered in 1983 in mouse erythroleukemia cells. Over the years, it became evident that TCTP is an important player in a number of basic cell physiology events in cancer, embryo development, cell cycle, apoptosis, proliferation, growth, stress response, allergy, gene regulation, and heat-shock response. However, despite the nearly three decades of research, we only start to understand the role of TCTP in physiology of animal and plant embryo development as well as in numerous pathologies through its participation in cell cycle, proliferation, and growth regulation. The exact roles of TCTP in many complex cellular processes still remain a mystery. One of the key questions in cancer research is the role of TCTP in tumor reversion, the rare event leading to tumor regression and a “miraculous” cure: is TCTP involved in gene regulation or rather modification of the cytoskeleton of cancer cells during this process? It seems plausible that a novel type of posttranslational modification of TCTP, such as SUMOylation, by regulating its nuclear localization and/or its association with the centrosomes (both subjects featured in this issue) is responsible for some of the TCTP functions in normal and cancer cells. From presented in this issue very comprehensive and up-to-date reviews on TCTP functions, it clearly transpires that TCTP has a potential to be a crucial target for anticancer therapies. However, more research on the regulation of TCTP and its involvement in various molecular and cellular pathways and its association with subcellular structures is needed for the improvement of our understanding of this oncogene and the development of novel TCTP-targeted cancer therapies. We hope that our special TCTP issue will help in stimulation of scientific research in this field

Phosphorylated ERK5/BMK1 transiently accumulates within division spindles in mouse oocytes and preimplantation embryos

MAP kinases of the ERK family play important roles in oocyte maturation, fertilization, and early embryo development. The role of the signaling pathway involving ERK5 MAP kinase during meiotic and mitotic M-phase of the cell cycle is not well known. Here, we studied the localization of the phosphorylated, and thus potentially activated, form of ERK5 in mouse maturing oocytes and mitotically dividing early embryos. We show that phosphorylation/dephosphorylation, i.e. likely activation/inactivation of ERK5, correlates with M-phase progression. Phosphorylated form of ERK5 accumulates in division spindle of both meiotic and mitotic cells, and precisely co-localizes with spindle microtubules at metaphase. This localization changes drastically in the anaphase, when phospho-ERK5 completely disappears from microtubules and transits to the cytoplasmic granular, vesicle-like structures. In telophase oocytes it becomes incorporated into the midbody. Dynamic changes in the localization of phospho-ERK5 suggests that it may play an important role both in meiotic and mitotic division. (Folia Histochemica et Cytobiologica 2011, Vol. 49, No. 3, 528–534

Developmental Herpetology - state of the art of amphibian and reptile developmental biology

International audienceThe Special Issue which you are now reading is the offspring of a vivid backstage conversation during a scientific meeting with the Editor-in-Chief of The International Journal of Developmental Biology (Int. J. Dev. Biol.) on the importance of research on the developmental biology of Amphibians and Reptiles yesterday, today and in the future. As you can see, we managed to convince the Editor-in-Chief that the matter is indeed important. We hope you will enjoy the outcome

Cytostatic factor inactivation is induced by a calcium-dependent mechanism present until the second cell cycle in fertilized but not in parthenogenetically activated mouse eggs

Cytostatic factor (CSF) is an activity responsible for the metaphase II arrest in vertebrate oocytes. This activity maintains a high level of maturation promoting factor (MPF) in the oocyte and both activities are destroyed after fertilization or parthenogenetic activation. To study some of the characteristics of the mechanism involved in MPF and CSF destruction, we constructed hybrid cells between metaphase II arrested oocytes and early embryos obtained after fertilization or artificial activation. We found that the behavior of hybrid cells differed depending upon the type of oocyte activation. Initially, the reaction of both types of hybrid cells was similar, the nuclear envelope broke down and chromatin condensation was induced. However, while metaphase II oocytes fused with parthenogenetic eggs remained arrested in M-phase, the oocytes fused with fertilized eggs underwent activation and passed into interphase. This ability of fertilized eggs to induce oocyte activation was still present at the beginning, but not at the end of the second embryonic cell cycle. Oocyte activation induced by fusion with a fertilized egg could be prevented when calcium was chelated by BAPTA. Thus, element(s) of the mechanism involved in calcium release triggered by a sperm component at fertilization remain(s) active until the second cell cycle and is (are) inactivated before the end of the 2-cell stage

Proteomics reveals a switch in CDK1-associated proteins upon M-phase exit during the Xenopus laevis oocyte to embryo transition.

International audienceCyclin-dependent kinase 1 (CDK1) is a major M-phase kinase which requires the binding to a regulatory protein, Cyclin B, to be active. CDK1/Cyclin B complex is called M-phase promoting factor (MPF) for its key role in controlling both meiotic and mitotic M-phase of the cell cycle. CDK1 inactivation is necessary for oocyte activation and initiation of embryo development. This complex process requires both Cyclin B polyubiquitination and proteosomal degradation via the ubiquitin-conjugation pathway, followed by the dephosphorylation of the monomeric CDK1 on Thr161. Previous proteomic analyses revealed a number of CDK1-associated proteins in human HeLa cells. It is, however, unknown whether specific partners are involved in CDK1 inactivation upon M-phase exit. To better understand CDK1 regulation during MII-arrest and oocyte activation, we immunoprecipitated (IPed) CDK1 together with its associated proteins from M-phase-arrested and M-phase-exiting Xenopus laevis oocytes. A mass spectrometry (MS) analysis revealed a number of new putative CDK1 partners. Most importantly, the composition of the CDK1-associated complex changed rapidly during M-phase exit. Additionally, an analysis of CDK1 complexes precipitated with beads covered with p9 protein, a fission yeast suc1 homologue well known for its high affinity for CDKs, was performed to identify the most abundant proteins associated with CDK1. The screen was auto-validated by identification of: (i) two forms of CDK1: Cdc2A and B, (ii) a set of Cyclins B with clearly diminishing number of peptides identified upon M-phase exit, (iii) a number of known CDK1 substrates (e.g. peroxiredoxine) and partners (e.g. HSPA8, a member of the HSP70 family) both in IP and in p9 precipitated pellets. In IP samples we also identified chaperones, which can modulate CDK1 three-dimensional structure, as well as calcineurin, a protein necessary for successful oocyte activation. These results shed a new light on CDK1 regulation via a dynamic change in the composition of the protein complex upon M-phase exit and the oocyte to embryo transition