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J Gautier

Publications and source records attributed to J Gautier.

At least 37 records · Page 2Linked to original sources
PubMed1999

Developmental regulation of induced and programmed cell death in Xenopus embryos.

We have analyzed the role of cell death during early Xenopus development and have identified two distinct types of cell death programs during the period between fertilization and the tadpole stage. One is a maternal cell death program that is activated at the onset of gastrulation following damage to the pre-midblastula transition embryo, resulting in the death of non-viable cells. The activation of this cell death program at a specific time during development is a maternally programmed event under the control of a developmental timer set at fertilization, and does not depend on the type of stress applied, on cell cycle progression, or de novo protein synthesis. Subsequently, a second program corresponding to programmed cell death is initiated as part of the normal development of the embryo. Programmed cell death starts at the onset of gastrulation and we have analyzed its spatio-temporal patterns by a whole-mount in situ DNA end labeling technique (the TUNEL protocol).

Animals↗
PubMed1999

Does spinal morphology influence the occurrence of low back pain? A retrospective clinical, anthropometric, and radiological study.

OBJECTIVE: Insurance companies are showing considerable interest for measurement devices claimed to be effective in evaluating risks associated with occupational or sporting activities. The aim of this study was to evaluate the potential influence of spinal morphology on the risk of low back pain. METHODS: 1500 male construction work apprentices aged 16 to 23 years underwent spinal measurements at the Caen Teaching Hospital from 1991 to 1996. The 384 subjects with abnormal measurements or a history of spinal pain underwent plain radiography and a second set of spinal measurements done by a different examiner using a different device. The following radiological parameters were studied: incidence angle, junction couple, Ferguson's index, L1-L5 lordosis angle and L1-S1 lordosis angle. RESULTS: In the 152 retrospectively studied subjects, no significant differences in static spinal parameters were found between subjects with and without a history of low back pain. The type of lordosis was correlated with the incidence angle but not with the junction couple. CONCLUSION: In our population, spinal measurements were not correlated with a specific morphological pattern of the lumbar spine or pelvis, and neither was a history of low back pain correlated with a specific pattern of lumbar and pelvic balance.

Adolescent↗
PubMed1998

Programmed cell death during Xenopus development: a spatio-temporal analysis.

Programmed cell death (PCD) is an integral part of many developmental processes. In vertebrates little is yet known on the patterns of PCD and its role during the early phases of development, when embryonic tissue layers migrate and pattern formation takes place. We describe the spatio-temporal patterns of cell death during early Xenopus development, from fertilization to the tadpole stage (stage 35/36). Cell death was analyzed by a whole-mount in situ DNA end-labeling technique (the TUNEL protocol), as well as by serial sections of paraffin-embedded TUNEL-stained embryos. The first cell death was detected during gastrulation, and as development progressed followed highly dynamic and reproducible patterns, strongly suggesting it is an important component of development at these stages. The detection of PCD during neural induction, neural plate patterning, and later during the development of the nervous system highlights the role of PCD throughout neurogenesis. Additionally, high levels of cell death were detected in the developing tail and sensory organs. This is the first detailed description of PCD throughout early development of a vertebrate, and provides the basis for further studies on its role in the patterning and morphogenesis of the embryo.

Animals↗
PubMed1998

Developmental regulation of MCM replication factors in Xenopus laevis.

At the midblastula transition (MBT) during Xenopus laevis development, zygotic transcription begins [1], and the rapid, early cleavage cycles are replaced by cell-division cycles that lengthen and acquire G (gap) phases [2] and checkpoints [3-5]. This cell-cycle remodeling may result from either a loss of maternal products, the transcription of zygotic genes, or the replacement of maternal proteins by zygotic gene products. We have identified an example of the third possibility: distinct maternal and zygotic genes encoding a member of the minichromosome maintenance (MCM) protein family. The mcm genes were identified in yeast by mutations that blocked replication of artificial chromosomes or perturbed the G1/S transition in the cell cycle [6,7]. In Xenopus eggs, the MCM2-MCM7 proteins assemble as multimeric complexes at chromosomal origins of replication [8-14]. The sequential, cell-cycle-dependent assembly of the origin replication complex (ORC), CDC6 protein and the MCM complex at origins of replication ensures that DNA replicates only once per cell cycle [15,16]. The periodic association of the MCM complex with chromatin may be regulated via phosphorylation by cyclin-dependent kinases (Cdks) [11]. We have cloned the first example of a developmentally regulated mcm gene, zygotic mcm6 (zmcm6), expressed only after gastrulation when the cell cycle is remodeled. The zMCM6 protein assembles into MCM complexes and differs from maternal MCM6 (mMCM6) in having a carboxy-terminal extension and a consensus cyclin-Cdk phosphorylation site. There may also be maternal-zygotic pairs of other MCMs. These data suggest that MCMs are critical for cell-cycle remodeling during early Xenopus development.

Animals↗
PubMed1997

A developmental timer that regulates apoptosis at the onset of gastrulation.

Recent work identified an apoptotic program in gastrulation stage Xenopus embryos (Anderson, J.A., Lewellyn, A.L., Maller, J.L., 1997. Mol. Biol. Cell 8, 1195-1206; Stack, J.H., Newport, J.W., 1997. Development 124, 3185-3195). Here, we characterize in detail this maternal cell death program, which is set up at fertilization and abruptly activated at the onset of gastrulation, following DNA damage or treatment of embryos with inhibitors of transcription, translation, or replication, between the time of fertilization and the midblastula transition (MBT). This apoptotic pathway is activated under tightly regulated developmental control(s): if the same treatments are applied after the MBT the apoptotic response is abrogated. Embryos displayed many characteristic apoptotic features, including DNA fragmentation, caspase activation, and embryonic death was blocked in vivo by the ectopic expression of Bcl-2, or injection of the caspase-3 inhibitor z-DEVD-fmk. The precise timing and the execution of this maternal cell death program is set at fertilization and does not depend on the type of stress applied, on cell cycle progression, or on de novo protein synthesis. This maternal developmental program might palliate the lack of cell cycle checkpoints in the pre-MBT embryo.

Animals↗
PubMed1996

Phosphorylation of MCM4 by cdc2 protein kinase inhibits the activity of the minichromosome maintenance complex.

In eukaryotes, tight regulatory mechanisms ensure the ordered progression through the cell cycle phases. The mechanisms that prevent chromosomal DNA replication from taking place more than once each cell cycle are thought to involve the function of proteins of the minichromosome maintenance (MCM) family. Here, we demonstrate that Xenopus MCM4, a member of the MCM protein family related to Spcdc21/ ScCDC54, is part of a large protein complex comprising several other MCM proteins. MCM4 undergoes cell cycle-dependent phosphorylation both in cleaving embryos and in cell-free extracts. MCM4 phosphorylation starts concomitantly with the clearing of the MCM complex from the chromatin during S phase. Phosphorylation is carried out by cdc2/cyclinB protein kinase, which phosphorylates MCM4 in vitro at identical sites as the ones phosphorylated in vivo. Phosphorylation is specific for cdc2 protein kinase since MCM4 is not a substrate for other members of the cdk family. Furthermore, phosphorylation of MCM4 dramatically reduces its affinity for the chromatin. We propose that the cell cycle-dependent phosphorylation of MCM4 is a mechanism which inactivates the MCM complex from late S phase through mitosis, thus preventing illegitimate DNA replication during that period of the cell cycle.

Animals↗
PubMed1995

Regulation of cell cycle progression following DNA damage.

DNA damage causes an arrest in cell cycle progression. Checkpoints, which monitor the state of the DNA, exist throughout the cycle and negatively regulate cell cycle transitions when damage is detected. The molecular basis of how these checkpoints are activated, and interact with the cell cycle machinery, is just beginning to be understood. Studies in yeast have identified a number of genes involved in a G2 DNA damage checkpoint, while in mammalian cells a G1 checkpoint has been extensively studied.

Cell Cycle↗
PubMed1995

Transluminal recanalization of occluded iliac arteries: a surgical experience.

The purpose of this study was to evaluate the feasibility of transluminal techniques in an unselected group of patients and to assess long-term outcome in successful procedures. All patients in whom iliac artery recanalization was attempted were included in this study. Patients with an occluded prosthesis or recent embolism were excluded. A total of 37 patients were studied. Assessment of the success or failure of the procedure was based on the results of control arteriography. Recanalization was deemed successful in 31 patients. This study demonstrates that transluminal recanalization of occluded iliac arteries by a surgeon is indeed feasible. Primary patency was 66% at 24 months. The potential risk, however, is long-term restenosis. Thus follow-up examination every 6 months is recommended including pressure index measurements after exercise and color Doppler ultrasonography of the recanalized zone.

Adult↗
PubMed1992

A cdc2-related kinase oscillates in the cell cycle independently of cyclins G2/M and cdc2.

The Eg1 gene in Xenopus laevis is related in sequence to the cdc2+ gene. We show here that the Eg1 gene product (cdk2) possesses histone H1 protein kinase activity and binds to PSTAIR antibodies as well as to Sepharose beads linked to the 13-kDa product of the suc 1 gene (p13suc1). Eg1 protein kinase is active only in an Mr approximately 200,000 complex with other proteins but is not associated with any of the three known Xenopus mitotic cyclins or with any newly synthesized protein in egg extracts that exhibit cell cycle oscillations in vitro. The protein kinase activity of Eg1 oscillates in the mitotic cell cycle, being high in M-phase and low in interphase. Hyperactivation of cdc2 kinase by the addition of cyclin A has no effect on the activity or oscillatory behavior of Eg1. Inhibition of cdc2 kinase activation by emetine or RNase treatment of oscillating extracts does not inhibit the activation of Eg1 but does block deactivation normally seen during exit from mitosis. These results indicate that Eg1 is regulated by a cell cycle clock independently of cyclin and cdc2 kinase.

Amino Acid Sequence↗
PubMed1991

cdc25 is a specific tyrosine phosphatase that directly activates p34cdc2.

cdc25 controls the activity of the cyclin-p34cdc2 complex by regulating the state of tyrosine phosphorylation of p34cdc2. Drosophila cdc25 protein from two different expression systems activates inactive cyclin-p34cdc2 and induces M phase in Xenopus oocytes and egg extracts. We find that the cdc25 sequence shows weak but significant homology to a phylogenetically diverse group of protein tyrosine phosphatases. cdc25 itself is a very specific protein tyrosine phosphatase. Bacterially expressed cdc25 directly dephosphorylates bacterially expressed p34cdc2 on Tyr-15 in a minimal system devoid of eukaryotic cell components, but does not dephosphorylate other tyrosine-phosphorylated proteins at appreciable rates. In addition, mutations in the putative catalytic site abolish the in vivo activity of cdc25 and its phosphatase activity in vitro. Therefore, cdc25 is a specific protein phosphatase that dephosphorylates tyrosine and possibly threonine residues on p34cdc2 and regulates MPF activation.

Amino Acid Sequence↗
PubMed1991

Cyclin B in Xenopus oocytes: implications for the mechanism of pre-MPF activation.

Using a polyclonal antibody raised against B2 cyclin from Xenopus laevis, we show that prophase-arrested Xenopus oocytes contain a stockpile of cyclin B2 protein. During progesterone-induced maturation, an increase in the synthesis of cyclin B2 is observed, although Western blotting experiments show that this new synthesis does not significantly increase the mass of cyclin over the maternal stockpile. In the oocyte cyclin B2 is already present in two forms which differ in the extent of phosphorylation, but the phosphorylated form becomes predominant as oocytes progress towards germinal vesicle breakdown (GVBD), coincident with cdc2 protein kinase activation. These two events do not depend upon formation of a new complex between cyclin and cdc2 protein kinase, since these two proteins are already found associated in resting oocytes, prior to activation of the kinase.

Animals↗
PubMed1990

The cyclin B2 component of MPF is a substrate for the c-mos(xe) proto-oncogene product.

Previous studies from this laboratory have shown that purified MPF from Xenopus eggs contains cyclin B2 complexed with cdc2 kinase. The activation of MPF during oocyte maturation is known to require expression of the c-mos(xe) proto-oncogene. We show here that immunoprecipitates of either v-mos from Moloney murine sarcoma virus-transformed NIH 3T3 cells or c-mos from Xenopus eggs phosphorylate cyclin B2 in vitro. Phosphopeptide analysis reveals a pattern similar to that observed with cdc2 kinase. Moreover, ablation of c-mos(xe) from oocytes by antisense oligonucleotide injection reduces the rate of cyclin B2 phosphorylation in oocyte extracts by 40%. These results suggest that the mechanism of activation of MPF by c-mos(xe) involves phosphorylation of the cyclin component.

Animals↗
PubMed1990

Cyclin is a component of maturation-promoting factor from Xenopus.

Highly purified maturation-promoting factor (MPF) from Xenopus eggs contains both cyclin B1 and cyclin B2 as shown by Western blotting and immunoprecipitation using Xenopus anti-B-type cyclin antibodies. Immunoprecipitates with these antibodies display the histone H1 kinase activity characteristic of MPF, for which exogenously added B1 and B2 cyclins are both substrates. Protein kinase activity against cyclin oscillates in maturing oocytes and activated eggs with the same kinetics as p34cdc2 kinase activity. These data indicate that B-type cyclin is the other component of MPF besides p34cdc2.

Adenosine Triphosphate↗
PubMed1989

Dephosphorylation and activation of Xenopus p34cdc2 protein kinase during the cell cycle.

Genetic studies in the fission yeast Schizosaccharomyces pombe have established that a critical element required for the G2----M-phase transition in the cell cycle is encoded by the cdc2+ gene. The product of this gene is a serine/threonine protein kinase, designated p34cdc, that is highly conserved functionally from yeast to man2 and has a relative molecular mass of 34,000 (34 K). Purified maturation-promoting factor (MPF) is a complex of p34cdc2 and a 45K substrate that appears in late G2 phase and is sufficient to drive cells into mitosis. This factor has been identified in all eukaryotic cells, and in vitro histone H1 is the preferred substrate for phosphorylation. The increase in the activity of H1 kinase in M-phase is associated with a large increase in total cell protein phosphorylation which is believed to be a consequence of MPF activation. We show here that the H1 kinase activity of p34cdc2 oscillates during the cell cycle in Xenopus, and maximal activity correlates with the dephosphorylated state of p34cdc2. Direct inactivation of MPF in vitro is accompanied by phosphorylation of p34cdc2 and reduction of its protein kinase activity.

Animals↗
PubMed1989

Mammalian growth-associated H1 histone kinase: a homolog of cdc2+/CDC28 protein kinases controlling mitotic entry in yeast and frog cells.

Mammalian growth-associated H1 histone kinase, an enzyme whose activity is sharply elevated at mitosis, is similar to cdc2+ protein kinase from Schizosaccharomyces pombe and CDC28 protein kinase from Saccharomyces cerevisiae with respect to immunoreactivity, molecular size, and specificity for phosphorylation sites in H1 histone. Phosphorylation of specific growth-associated sites in H1 histone is catalyzed by yeast cdc2+/CDC28 kinase, as shown by the in vitro thermal lability of this activity in extracts prepared from temperature-sensitive mutants. In addition, highly purified Xenopus maturation-promoting factor catalyzes phosphorylation of the same sites in H1 as do the mammalian and yeast kinases. The data indicate that growth-associated H1 kinase is encoded by a mammalian homolog of cdc2+/CDC28 protein kinase, which controls entry into mitosis in yeast and frog cells. Since H1 histone is known to be an in vivo substrate of the mammalian kinase, this suggests that phosphorylation of H1 histone or an H1 histone counterpart is an important component of the mechanism for entry of cells into mitosis.

Animals↗
PubMed1989

Maturation-promoting factor and the regulation of the cell cycle.

Maturation-promoting factor (MPF) is a cell cycle control element able to cause metaphase when injected into amphibian oocytes or when incubated with nuclei in a cell-free system. Highly purified MPF consists of a complex between a 34K (K = 10(3) Mr) serine/threonine protein kinase, identified as a Xenopus homolog of the cdc2+ gene product, p34cdc2, and a 45K substrate, identified as a Xenopus B-type cyclin. p34cdc2 is also present in purified preparations of chromatin-derived growth-associated histone H1 kinase from Novikoff hepatoma cells. p34cdc2 is active when dephosphorylated and inactive when phosphorylated during oocyte meiotic cell cycles and in mitotic cell cycles following egg activation. Analysis of the substrate specificity of p34cdc2 indicates a consensus sequence for phosphorylation of (K/R)S/TP(X)K/R. Among substrates identified with this consensus are histone H1 and the pp60c-src proto-oncogene, which is known to be activated and phophorylated in mitosis. MPF injection into oocytes activates ribosomal protein S6 kinase II, which is also a lamin kinase. The mechanism of activation is indirect, possibly involving the c-src proto-oncogene. Continued analysis of regulation of MPF activation/inactivation and characterization of substrates for phosphorylation will have important implications for cell cycle and cell growth control.

Animals↗