PITC derivatives in amino acid analysis

Refined methods for separating PTC-amino acids on reverse phase columns may pose a challenge to traditional ion exchange techniques.     

Meiotic arrest and aneuploidy in MLH3-deficient mice

MutL homolog 3 (Mlh3) is a member of a family of proteins conserved during evolution and having dual roles in DNA mismatch repair and meiosis. The pathway in eukaryotes consists of the DNA-binding components, which are the homologs of the bacterial MutS protein (MSH 2 6), and the MutL homologs, which bind to the MutS homologs and are essential for the repair process. Three of the six homologs of MutS that function in these processes, Msh2, Msh3 and Msh6, are involved in the mismatch repair of mutations, frameshifts and replication errors, and two others, Msh4 and Msh5, have specific roles in meiosis. Of the four MutL homologs, Mlh1, Mlh3, Pms1 and Pms2, three are involved in mismatch repair and at least two, Pms2 and Mlh1, are essential for meiotic progression in both yeast and mice. To assess the role of Mlh3 in mammalian meiosis, we have generated and characterized Mlh3(-/-) mice. Here we show that Mlh3(-/-) mice are viable but sterile. Mlh3 is required for Mlh1 binding to meiotic chromosomes and localizes to meiotic chromosomes from the mid pachynema stage of prophase I. Mlh3(-/-) spermatocytes reach metaphase before succumbing to apoptosis, but oocytes fail to complete meiosis I after fertilization. Our results show that Mlh3 has an essential and distinct role in mammalian meiosis.     

Propagating solitary waves along a rapidly moving crack front

A rapidly moving crack in a brittle material is often idealized as a one-dimensional object with a singular tip, moving through a two-dimensional material. However, in real three-dimensional materials, tensile cracks form a planar surface whose edge is a rapidly moving one-dimensional singular front. The dynamics of these fronts under repetitive interaction with material inhomogeneities (asperities) and the morphology of the fracture surface that they create are not yet understood. Here we show that perturbations to a crack front in a brittle material result in long-lived and highly localized waves, which we call 'front waves' These waves exhibit a unique characteristic shape and propagate along the crack front at approximately the Rayleigh wave speed (the speed of sound along a free surface). Following interaction, counter-propagating front waves retain both their shape and amplitude. They create characteristic traces along the fracture surface, providing cracks with both inertia and a new mode of dissipation. Front waves are intrinsically three-dimensional, and cannot exist in conventional two-dimensional theories of fracture. Because front waves can transport and distribute asperity-induced energy fluctuations throughout the crack front, they may help to explain how cracks remain a single coherent entity, despite repeated interactions with randomly dispersed asperities.     

High brain densities of the immunophilin FKBP colocalized with calcineurin.

The immunophilins cyclophilin and FK506 binding protein (FKBP) are small, predominantly soluble proteins that bind the immunosuppressant drugs cyclosporin A and FK506, respectively, with high affinity, and which seem to mediate their pharmacological actions. The Ca(2+)-dependent protein phosphatase, calcineurin, binds the cyclophilin-cyclosporin A and FKBP-FK506 complexes, indicating that calcineurin might mediate the actions of these drugs. A physiological role for the immunophilins in the nervous system is implied by a close homology between the structure of NINA A, a protein in the neural retina of Drosophila, and cyclophilin, as well as by the high density of FKBP messenger RNA in brain tissue. Here we report that the levels of FKBP and mRNA in rat brain are extraordinarily high and that their regional localization is virtually identical to that of calcineurin, indicating that there may be a physiological link between calcineurin and the immunophilins. We also show that at low concentrations FK506 and cyclosporin A enhance the phosphorylation of endogenous protein substrates in brain tissue and in intact PC12 cells, indicating that these drugs may inhibit phosphatase activity by interacting with the immunophilin-calcineurin complexes.     

Dissection of the protein kinase cascade by which nerve growth factor activates MAP kinases.

Mitogen activated protein (MAP) kinases (MAPKs) are a family of protein-serine/threonine kinases activated as an early intracellular response to a variety of hormones and growth factors. They are unique in requiring both serine/threonine and tyrosine phosphorylation to become active and are the only examples of protein-serine/threonine kinases activated by tyrosine phosphorylation. Nerve growth factor (NGF) promotes differentiation of phaeochromocytoma (PC12) cells, which respond by conversion within hours from a chromaffin-like to a sympathetic neuron-like phenotype. NGF stimulation of PC12 cells increases the activity of two protein kinases by greater than 20-fold within minutes, both strikingly similar to MAPKs. They are inactivated by either protein-tyrosine phosphatases or the protein-serine/threonine phosphatase termed protein phosphatase 2A (ref. 8), they activate protein S6 kinase-II (refs 9, 10), and they phosphorylate identical threonine residues on myelin basic protein (our unpublished results) to those phosphorylated by other MAPKs. Immunological data indicate that these protein kinases, termed peak-I and peak-II (Fig. 1a) are probably ERK2 and ERK1, respectively, two widely expressed MAPK isoforms. Here we identify the 'MAP kinase kinases' (MAPKKs) in PC12 cells which are activated by NGF and report that MAPKKs are dependent on serine/threonine phosphorylation for activity and promote phosphorylation of serine/threonine and tyrosine residues on MAPKs.     

RandomX-chromosome inactivation in interspecific hybrids ofMeriones libycus (♂) ×Meriones shawi (♀) (Rodentia: Gerbillinae)

Résumé Des cultures obtenues à partir d'hybrides femelles entre deux espèces deMeriones à 44 chromosomes et différant par l'acrocentrie (M. shawi) ou la métacentrie (M. libycus) de l'X ont permis l'étude de clones cellulaires. C'est alors tantôt l'X métacentrique, tantôt l'X acrocentrique qui se révèle inactivé («latereplicating»). Bien que la proportion 1/1, significative d'une inactivation due uniquement au hasard, n'ait pas été rigoureusement observée, ces résultats sont nettement en faveur de l'hypothèse deLyon.

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Supported by Project No. 417 from the U.S. Children's Bureau and National Science Foundation (Grant No. GB 5676X).     

The role of protein phosphorylation in neural and hormonal control of cellular activity

Protein phosphorylation is now recognized to be the major general mechanism by which intracellular events in mammalian tissues are controlled by external physiological stimuli. However, only recently has the idea that different cellular functions are controlled by common protein kinases and protein phosphatases started to gain widespread acceptance. Thus there is an integrated network of regulatory pathways, mediated by phosphorylation-dephosphorylation, that allows diverse cellular events to be coordinated by neural and hormonal stimuli. The evidence that supports this concept is reviewed, with emphasis on the role of protein phosphorylation in enzyme regulation.     

Half-life of dehydroepiandrosterone sulfate (DHAS) during pregnancy]

During pregnancy the half-life of DHAS was measured after non tritiated DHAS infusion. At 30 weeks, the mean value obtained for DHAS half-life was 3.64 h +/- 0.74 and it was 3.67h +/- 0.38 at 38 weeks. The half-life of DHAS is shorter in pregnant than in non-pregnant women. Among 14 pathological pregnancies that we studied, 13 cases exhibited a DHAS half-life significantly longer than the average value obtained in normal pregnancies. In retarded fetal growth, the half-life of DHAS was more closely related to fetal than to placental weight.