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Biomedical subjects

A Sobel

Publications and source records attributed to A Sobel.

At least 55 records · Page 3Linked to original sources
PubMed1997

KIS is a protein kinase with an RNA recognition motif.

Protein phosphorylation is involved at multiple steps of RNA processing and in the regulation of protein expression. We present here the first identification of a serine/threonine kinase that possesses an RNP-type RNA recognition motif: KIS. We originally isolated KIS in a two-hybrid screen through its interaction with stathmin, a small phosphoprotein proposed to play a general role in the relay and integration of diverse intracellular signaling pathways. Determination of the primary sequence of KIS shows that it is formed by the juxtaposition of a kinase core with little homology to known kinases and a C-terminal domain that contains a characteristic RNA recognition motif with an intriguing homology to the C-terminal motif of the splicing factor U2AF. KIS produced in bacteria has an autophosphorylating activity and phosphorylates stathmin on serine residues. It also phosphorylates in vitro other classical substrates such as myelin basic protein and synapsin but not histones that inhibit its autophosphorylating activity. Immunofluorescence and biochemical analyses indicate that KIS overexpressed in HEK293 fibroblastic cells is partly targetted to the nucleus. Altogether, these results suggest the implication of KIS in the control of trafficking and/or splicing of RNAs probably through phosphorylation of associated factors.

Amino Acid Sequence↗
PubMed1997

Stathmin: a tubulin-sequestering protein which forms a ternary T2S complex with two tubulin molecules.

Stathmin is an important regulatory protein thought to control the dynamics of microtubules through the cell cycle in a phosphorylation-dependent manner. Here we show that stathmin interacts with two molecules of dimeric alphabeta-tubulin to form a tight ternary T2S complex, sedimenting at 7.7 S. This complex appears in slow association-dissociation equilibrium in the analytical ultracentrifuge. The T2S complex is formed under a variety of ionic conditions, either from GTP- or GDP-tubulin or from the tubulin-colchicine complex. The S16/25/38/63E mutated stathmin in contrast is in rapid equilibrium with tubulin in the T2S complex. The T2S complex cannot polymerize in microtubules nor in ring oligomers. Stathmin acts as a pure tubulin-sequestering protein via formation of the T2S complex. It does not act directly on microtubule ends to promote catastrophe nor enhance microtubule dynamics.

Animals↗
PubMed1997

The cytosolic phosphoprotein stathmin is expressed in the olfactory system of the adult rat.

Stathmin is a cytosolic protein expressed particularly in the developing nervous system, whose phosphorylation is correlated with the action of multiple extracellular stimuli regulating cell proliferation and differentiation. In this study, we used an antibody that specifically recognizes the carboxyterminal region of stathmin to analyze the distribution of this protein in the olfactory system of adult rats, and found a high and selective immunoreactivity in immature olfactory receptors of the olfactory neuroepithelium and in cells of the rostral migratory stream. These results reveal an expression of stathmin in regions of the adult nervous system characterized by striking structural plasticity and cell renewal, suggesting that this protein could play a role in the differentiation of newly generated cell populations.

Animals↗
PubMed1997

[Observance in the matter of health].

Despite the common-sense nature of ordinary rules of "good health" and general acceptance of scientific knowledge, non-compliance to medical prescriptions is frequent. In a survey conducted in 1994, the French Committee on Education and Health observed a 59.5% rate of self-prescription. In addition, 57.9% of all physician's prescriptions are modified by the patient. Most patients do not really question the competence of the prescriber nor the pertinence of the prescribed treatment: their non-compliance is simply an expression of their desire to "adapt" the prescription to their own personal needs. This classical problem of non-compliance to medical prescriptions takes on a much more dramatic tone in case of HIV infection. For the individual patient, the risk concerns not only his own health status, but also his conception of personal liberty and self-identify. For the general public and health authorities, the risk concerns disease spread, health care costs, viral mutations, resistance to treatment and production of scientific knowledge. In their article in this issue of La Presse Médicale, N. Munzenberger et al. address this problem in an analysis of patient compliance during an HIV treatment trial. They observed that clinical research in HIV is highly influenced by social interrelationships due to the different or contradictory personal objectives of the participants (prescribers and patients). Thus it is essential for those who design prospective protocols to take into account social relationships and manage clinical trials within the real social environment. Finally, the results reported underline the importance of reconsidering the myth of a "double-blind" protection against bias and the true meaning of "informed consent".

Clinical Trials as Topic↗
PubMed1996

Growth and cell density-dependent expression of stathmin in C2 myoblasts in culture.

Stathmin is a 19-kDa, ubiquitous cytoplasmic phosphoprotein whose expression is strongly regulated during tissue development and maturation and which was proposed as a general relay integrating diverse intracellular signaling pathways. Since myoblasts tend to align and differentiate in vitro toward myotubes above a certain density in culture, we examined the expression of stathmin as a function of cell density in the C2 myogenic cell line. Whereas stathmin was hardly detectable in low-to medium-density cultures corresponding to less than 1 microgram soluble protein/cm2, it became expressed to a stable level above this threshold of cell density. This cell density effect on stathmin expression was not mediated by a diffusible factor, since myogenic C2 or fibroblastic 3T3 cells grown at low and high density within the same culture flask displayed the same pattern of density-dependent stathmin expression, significant stathmin levels being observed only in the dense moiety of the flask. Interestingly, culture conditions which indirectly perturb cell-cell contacts, such as low Ca2+ or incubation with cytoskeleton disrupting agents such as nocodazole or cytochalasin D, prevented the expression of stathmin in C2 cells even at high density. More directly, anti-E-cadherin immunoglobulins, interfering with direct cell-cell contacts of the E-cadherin expressing S180 sarcoma-derived 2B2 cells, also prevented the expression of stathmin in these cells even at high density. Altogether, our results indicate that cell-cell contacts, probably mediated by adhesion molecules such as cadherins, are responsible for the high-density-induced expression of stathmin, which might then participate, in particular in the case of myogenic cells, in the control of the proliferation of cells and of their entry into the differentiation process.

Animals↗
PubMed1996

Identification and molecular characterization of Unc-33-like phosphoprotein (Ulip), a putative mammalian homolog of the axonal guidance-associated unc-33 gene product.

The control of neuritic extension and guidance is critical for the development, maturation, and regeneration of functional neuronal circuits. We identified a neuronal 64-85 kDa phosphoprotein, the expression of which in mouse brain is regulated during development, reaching a peak at approximately 5 d postnatal, when maturation of neurons and synaptic connections is highly active. The amino acid sequence of the mouse protein deduced from its cloned cDNA reveals similarities with that of the neuritic outgrowth- and guidance-related product of the unc-33 gene in Caenorhabditis elegans. The regulation of its phosphorylation in response to nerve growth factor, as well as its localization in neurites and growth cones and at the neuromuscular junction, further indicates that Ulip (for Unc-33-like phosphoprotein) is not only a structural but likely is also a functional mammalian homolog of Unc-33, potentially involved in the control of neuritic outgrowth and axonal guidance.

Animals↗
PubMed1996

[Selected clinical characteristics of insulin dependent diabetes (IDDM) discovered in the years 1990-1992 in patients 0-29 years of age from the Rzeszów voivodship].

The aim of the study was to analyse the clinical symptoms preceding the final diagnosis of IDDM, some features of the illness at the moment of the diagnosis and variants of treatment. We examined 54 patients, among them 28 males and 26 females. There were 23 and 31 persons between 0-14 and 15-29 years of age, respectively. The data were accumulated using questionnaires. We have shown that the newly diagnosed IDDM cases were characterised by typical clinical symptoms, early detection and 100% hospitalization. Most of the patients or their parents underwent the diabetic education during the patients first stay at hospital. Only monocomponent or human insulins were used in the case of patients between 0-14 years of age. Among the group 15-29 years of age in 13% of patients ordinary insulins (regular, NPH, Lente group) were administered as well. Intensive insulin therapy was used exclusively, but sporadically in the patients 15-29 years of age.

Adolescent↗
PubMed1995

Stathmin interaction with a putative kinase and coiled-coil-forming protein domains.

Stathmin is a ubiquitous, cytosolic 19-kDa protein, which is phosphorylated on up to four sites in response to many regulatory signals within cells. Its molecular characterization indicates a functional organization including an N-terminal regulatory domain that bears the phosphorylation sites, linked to a putative alpha-helical binding domain predicted to participate in coiled-coil, protein-protein interactions. We therefore proposed that stathmin may play the role of a relay integrating diverse intracellular regulatory pathways; its action on various target proteins would be a function of its combined phosphorylation state. To search for such target proteins, we used the two-hybrid screen in yeast, with stathmin as a "bait." We isolated and characterized four cDNAs encoding protein domains that interact with stathmin in vivo. One of the corresponding proteins was identified as BiP, a member of the hsp70 heat-shock protein family. Another is a previously unidentified, putative serine/threonine kinase, KIS, which might be regulated by stathmin or, more likely, be part of the kinases controlling its phosphorylation state. Finally, two clones code for subdomains of two proteins, CC1 and CC2, predicted to form alpha-helices participating in coiled-coil interacting structures. Their isolation by interaction screening further supports our model for the regulatory function of stathmin through coiled-coil interactions with diverse downstream targets via its presumed alpha-helical binding domain. The molecular and biological characterization of KIS, CC1, and CC2 proteins will give further insights into the molecular functions and mechanisms of action of stathmin as a relay of integrated intracellular regulatory pathways.

Amino Acid Sequence↗
PubMed1995

Stathmin is a major substrate for mitogen-activated protein kinase during heat shock and chemical stress in HeLa cells.

Stathmin is a ubiquitous, highly conserved 19-kDa cytoplasmic protein whose expression and phosphorylation are regulated in relation to cell proliferation, differentiation or activation, in many biological systems. In this report, we show that stathmin undergoes major phosphorylation in HeLa cells submitted to heat or chemical stress. Heat-shock-induced stathmin phosphorylation was very rapid, as maximal incorporation of phosphate was observed at 5 min. Phosphorylation of stathmin might, therefore, occur as a very early step in the intracellular response to heat shock. The sites of phosphorylation of stathmin involved during the stress response were identified as mostly Ser25 and, to a lesser extent, Ser38. These sites are both followed by a proline residue, and known to be good substrates in vitro for mitogen-activated protein kinase (MAP-kinase) and p34cdc2 kinase, respectively. In lysates from heat-shocked cells, an increased stathmin-kinase activity, distinct from the histone-H1-kinase activity, was found to phosphorylate stathmin mostly on Ser25, the main site for MAP-kinase in vitro. This stathmin-kinase coeluted quantitatively with the stress-activated MAP-kinase from an FPLC MonoQ column. Furthermore, a stathmin kinase activity was precipitated from lysates of heat-shocked HeLa cells by an anti-(MAP-kinase) serum. Together, these results indicate that the phosphorylation of stathmin by MAP-kinase is likely to be a significant component of the signalling array controlling the cellular response to stress, and they further underline the general involvement of stathmin in intracellular signalling.

Enzyme Activation↗
PubMed1995

The protein kinase C inhibitor H7 blocks phosphorylation of stathmin during TPA-induced growth inhibition of human pre-B leukemia REH6 cells.

The human pre-B acute lymphoblastic leukemia cell line REH6 was used to analyze the regulation of a ubiquitous intracellular phosphoprotein stathmin (Mr 19,000, pl = 5.6-6.2). We demonstrated by 32P-labeling that the short (1 h) treatment of the REH6 cells with the tumor promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), resulted in a rapid phosphorylation of at least three (P1, P2 and P3) stathmin isoforms without an alteration of stathmin isoform expression. Furthermore, Western blot analysis with specific antiserum showed that the prolonged period (48 h) of TPA treatment partially reduced protein levels particularly of two (N2 and P2) stathmin isoforms. The potent and relatively specific protein kinase C (PKC) inhibitor, 1,(5-isoquinolinesulphonyl)2methylpiperasine dihydrochloride (H7), partially inhibited these TPA effects, whereas the specific calmodulin inhibitor R24571 (calmidazolium) had no effect upon these events. Our findings suggest that stathmin phosphorylation in REH6 cells could be in part mediated by PKC activation.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1995

Stathmin gene expression in mammary gland and in Nb2 cells.

Mammary gland growth occurs essentially during pregnancy and induction of milk synthesis is triggered at parturition. Prolactin is mammogenic in vivo but only marginally in vitro. Prolactin induces milk synthesis in vivo and in cultured mammary cells. Prolactin is also strictly required for the multiplication of the rat lymphoid Nb2 cells. Stathmin is an ubiquitous and highly conserved phosphoprotein which seems to be involved in the intracellular mechanisms which trigger cell multiplication and differentiation. In the present study, the concentration of stathmin mRNA has been evaluated during the pregnancy-lactation-weaning cycle in mouse and rabbit. Stathmin mRNA appeared at its highest level during pregnancy and it was almost undetectable during lactation. Prolactin injected into mid-pregnant rabbits induced milk synthesis and this effect was not accompanied by any modification of stathmin mRNA concentration. In cultured primary rabbit mammary cells, prolactin induced casein gene expression without any alteration of stathmin mRNA concentration. In Nb2 cells, prolactin induced a progressive increase of stathmin mRNA concentration. This effect was not significant until after 4 h of prolactin action. These data suggest that stathmin is involved in mammary and Nb2 cell multiplication but may not be necessary for mammary cell differentiation.

Actins↗
PubMed1995

[Some aspects of epidemiology in insulin-dependent diabetes mellitus (IDDM) (I)].

Epidemiological studies based on registries from many centers in the world revealed geographical variation of IDDM, with the highest incidence in Scandinavian countries. There is a gradient north to south in occurrence of IDDM, and differences in incidence between urban and rural populations. Poland is a country with a rather low incidence of IDDM, not exceeding 6/100,000 in the population 0-29 years of age until 1992. Generally a higher incidence of IDDM has been observed in Whites, there are differences between migrating populations ethnically homogenous. The highest incidence occurs in 10-14 age group, with male sex predominance.

Adolescent↗
PubMed1995

[Some aspects of epidemiology in insulin-dependent diabetes mellitus (IDDM) (II)].

Seasonality is a characteristic feature of IDDM incidence. The lowest incidence is observed in summer, whereas the highest in winter/autumn. The risk of IDDM increases in children with independently: breast feeding shorter then 3 months, feeding with cow's milk instead of breast feeding, early introducing of solid diet in babies, or high consumption of cow's milk in later years. Children with new onset IDDM have elevated levels of the antibodies against cow's milk proteins and beta-lactoglobulin. It has been proved that there is a connection with HLA DQalfaArg52 i DQbeta-Asp57 and polymorphic region of insulin gene, and the IDDM susceptibility.

Animals↗
PubMed1994

Molecular characterization of human stathmin expressed in Escherichia coli: site-directed mutagenesis of two phosphorylatable serines (Ser-25 and Ser-63).

Stathmin, a probable relay protein possibly integrating multiple intracellular regulatory signals [reviewed in Sobel (1991) Trends Biochem. Sci. 16, 301-305], was expressed in Escherichia coli at levels as high as 20% of total bacterial protein. Characterization of the purified recombinant protein revealed that it had biochemical properties very similar to those of the native protein. It is a good substrate for both cyclic AMP-dependent protein kinase (PKA) and p34cdc2, on the same four sites as the native eukaryotic protein. As shown by m.s., the difference in isoelectric points from the native protein is probably due to the absence of acetylation of the protein produced in bacteria. C.d. studies indicate that stathmin probably contains about 45% of its sequence in an alpha-helical conformation, as also predicted for the sequence between residues 47 and 124 by computer analysis. Replacement of Ser-63 by alanine by in vitro mutagenesis resulted in a ten times less efficient phosphorylation of stathmin by PKA which occurred solely on Ser-16, confirming that Ser-63 is the major target of this kinase. Replacement of Ser-25, the major site phosphorylated by mitogen-activated protein kinase in vitro and in vivo, by the charged amino acid glutamic acid reproduced, in conjunction with the phosphorylation of Ser-16 by PKA, the mobility shift on SDS/polyacrylamide gels induced by the phosphorylation of Ser-25. This result strongly suggests that glutamic acid in position 25 is able to mimic the putative interactions of phosphoserine-25 with phosphoserine-16, as well as the resulting conformational changes that are probably also related to the functional regulation of stathmin.

Base Sequence↗
PubMed1994

Zidovudine therapy and HIV encephalitis: a 10-year neuropathological survey.

OBJECTIVE: To assess the effect of zidovudine on productive HIV infection of the brain. DESIGN: To correlate the incidence of HIV-specific neuropathology with zidovudine therapy. PATIENTS: We examined 192 AIDS cases neuropathologically; 97 had never been treated with zidovudine, 72 had received zidovudine for over 3 months and until death, 23 had their treatment terminated more than 1 month before death. RESULTS: The incidence of HIV encephalitis/HIV leukoencephalopathy (HIVE/HIVL) and of multinucleated giant cells (MGC) was significantly lower in patients who had received zidovudine than in those who had never received zidovudine. The yearly incidence of HIVE/HIVL increased between 1982 and 1987 probably because of improved survival, and decreased between 1987 and 1990 although the percentage of patients treated with zidovudine increased. Since 1991 the incidence of HIVE/HIVL and of MGC increased slightly. The percentage of patients treated with zidovudine until death decreased and that of patients whose treatment was terminated increased concomitantly. In 1989 and 1990, most patients whose treatment was terminated had MGC and HIVE/HIVL. In 1991 and 1992 this incidence decreased markedly, coinciding with the introduction of dideoxyinosine therapy. CONCLUSION: Zidovudine treatment significantly reduces the occurrence of productive HIV infection of the brain in AIDS. Discontinuing zidovudine therapy may favour the occurrence of HIV encephalitis. Substitution therapy with dideoxyinosine also appears to protect against HIV-specific brain pathology.

AIDS Dementia Complex↗
PubMed1994

[Maternal age at the moment of birth and occurrence of insulin dependent diabetes mellitus (IDDM) in the child].

The aim of the study was to determine whether advanced maternal age (over 35 years) on birth of a child had any influence on the incidence of IDDM in her offspring. The analysis has also taken into account the presence of other factors which may have some bearing on the incidence of the disease. They are the following: viral infections prior to the onset of IDDM, sequence of the affected child's birth in relation to other siblings, and family history of diabetes. The study included 243 persons who fell ill with IDDM at the age of 0-29 years in the period of January 1st 1980 to December 31st 1992 and the pair matched control group. They were all residents of the Rzeszów province at the onset of the disease. We have found that a positive family history of diabetes, viral infection especially epidemic parotitis, and parity are the factors which increase the incidence of IDDM. Advanced maternal age, over 35 years on child's birth, along with the above-mentioned factors, increase the incidence of IDDM.

Adolescent↗
PubMed1993

Stathmin: cellular localization of a major phosphoprotein in the adult rat and human CNS.

Stathmin is a ubiquitous, 19 kDa cytoplasmic protein the phosphorylation of which is associated with many cellular signaling pathways. It is particularly abundant in neurons and reaches a peak of expression in the neonatal period, although it remains highly expressed in the adult brain. In order to determine whether this abundant expression is associated with discrete cellular populations that are still at an immature stage during adulthood, as suggested by others, the cellular localization of stathmin was investigated in the adult rat and human central nervous system. Western blotting with a specific antiserum indicated that stathmin was ubiquitous in the brain and spinal cord but that its relative concentration varied up to 2.6 times between regions. To characterize the distribution of stathmin within the brain, its cellular localization was analyzed by immunocytochemistry. Highly immunoreactive neurons and oligodendrocytes were observed, and stathmin immunoreactivity was localized to the perikaryon and all processes, but not the nucleus. Most brain and spinal cord cell groups showed stathmin immunoreactivity, although the extent and intensity of labeling differed largely from one place to another. Particularly numerous stathmin-immunoreactive neuronal cell bodies were found in the pyriform, cingulate, and neocortex, as well as in many cholinergic nuclei of the basal forebrain and brainstem, in the medial thalamus, in various brainstem nuclei, in the dorsalmost layers of the spinal cord, and in brain areas lacking a blood-brain barrier to macromolecules. In addition to neuronal populations, stathmin-antibodies intensely labeled choroid plexuses. Many other brain regions exhibited moderate neuronal immunostaining. The distribution of stathmin-immunoreactive processes was in some areas relatively heterogeneous. Intense immunoreactivity was observed in some fiber tracts (corpus callosum, anterior commissure, inferior cerebellar peduncle, etc.) but was missing in others (internal capsule, posterior commissure, etc.). Some brain areas rich in immunoreactive neurons also displayed an intense immunoreactivity of the neuropile, whereas others contained either immunoreactive cells or fibers. In the human brain, stathmin immunostaining occurred in many areas, corresponding to those identified in the rat, with the exception of the cerebral cortex, the hippocampal fascia dentata, and the substantia nigra. The present results support our suggestion that, in addition to its involvement in cell proliferation and differentiation, stathmin may also be related to regulation of differentiated cell functions, as it appears to be a major signaling protein in widespread areas of the adult brain in both rat and human.

Aged↗