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P Russell

Publications and source records attributed to P Russell.

At least 235 records · Page 13Linked to original sources
PubMed1991

A fission yeast B-type cyclin functioning early in the cell cycle.

We have cloned a fission yeast gene, cig1+, encoding a 48 kd product that is most similar to cyclin B proteins. The cig1+ protein has a "cyclin box" approximately 40% identical to B-type cyclins of other species, but lacks the "destruction box" required for proteolysis of mitotic cyclins. Deletion of cig1+ had no observable effect on cell viability or progression through G2 or M phase, but instead caused a marked lag in the progression from G1 to S phase. G1 constituted approximately 70% of the cell cycle in cig1 deletion strains, as compared with less than 10% in cig1+ strains. Constitutive cig1+ overexpression was lethal, causing cessation of growth and arrest in G1. Expression of cig1+ failed to rescue an S. cerevisiae strain lacking CLN Start cyclins. Thus, cig1+ identifies a new class of B-type cyclin acting in G1 or S phase that appears to be functionally distinct from all previously described cyclin proteins.

Amino Acid Sequence↗
PubMed1991

Dephosphorylation and activation of a p34cdc2/cyclin B complex in vitro by human CDC25 protein.

Oocytes arrested in the G2 phase of the cell cycle contain a p34cdc2/cyclin B complex which is kept in an inactive form by phosphorylation of its p34cdc2 subunit on tyrosine, threonine and perhaps serine residues. The phosphatase(s) involved in p34cdc2 dephosphorylation is unknown, but the product of the fission yeast cdc25+ gene, and its homologues in budding yeast and Drosophila are probably positive regulators of the transition from G2 to M phase. We have purified the inactive p34cdc2/cyclin B complex from G2-arrested starfish oocytes. Addition of the purified bacterially expressed product of the human homologue of the fission yeast cdc25+ gene (p54CDC25H) triggers p34cdc2 dephosphorylation and activates H1 histone kinase activity in this preparation. We propose that the cdc25+ gene product directly activates the p34cdc2-cyclin B complex.

Amino Acid Sequence↗
PubMed1991

Deletion mutation in an eye lens beta-crystallin. An animal model for inherited cataracts.

The most prevalent proteins in the lens of the eye are called crystallins, and it is thought that aberrant crystallins may cause opacification of lens tissue. The Philly mouse, a strain with an inherited cataract, has an abnormal beta B2-crystallin, the principal beta-crystallin in the mouse. The cDNA that codes for the beta B2-crystallin protein has been cloned and sequenced from both the normal and the cataractous Philly mouse. The normal mouse beta B2 cDNA is 756 nucleotides in length with 618 nucleotides of open reading frame. An in-frame deletion of 12 nucleotides has occurred in the Philly mouse cDNA, which results in the loss of 4 amino acids. The sequence of the mutant beta B2 was analyzed against the reported structure of the normal bovine beta B2-crystallin determined by x-ray crystallography. The region, in which the deletion of the amino acids occurs near the COOH terminus, is essential for the formation of the tertiary structure of the beta B2-crystallin. The loss of these residues could explain the alterations that are seen with the Philly beta B2 protein and may account for the instability of the Philly beta B2 protein. This abnormal beta B2-crystallin may be the cause of the cataract in this animal.

Amino Acid Sequence↗
PubMed1991

Fission yeast p107wee1 mitotic inhibitor is a tyrosine/serine kinase.

The fission yeast wee1+ gene product is a dose-dependent, negative regulator of entry into mitosis. wee1+ encodes a protein of relative molecular mass 107,000 (Mr 107K), the C-terminal third of which has strong similarities with the serine/threonine protein kinase family. Here we report that p107wee1 immune complexes phosphorylate p107wee1 equally on serine and tyrosine residues, and also phosphorylate an exogenous substrate, angiotensin II, on tyrosine. Both kinase activities are attributable to p107wee1 because they are also observed when wee1+ is expressed in heterologous systems; both are abolished by a point mutation in the ATP-binding domain, and both behave like an asymmetric monomer of Mr114K on gel filtration and density-gradient centrifugation. Thus the wee1+ gene product is representative of a novel class of protein kinase that phosphorylates both serine and tyrosine residues.

Animals↗
PubMed1991

p80cdc25 mitotic inducer is the tyrosine phosphatase that activates p34cdc2 kinase in fission yeast.

We have investigated the mechanism by which fission yeast p80cdc25 induces mitosis. The in vivo active domain was localized to the C-terminal 23 kDa of p80cdc25. This domain produced as a bacterial fusion protein (GST-cdc25) caused tyrosyl dephosphorylation and activation of immunoprecipitated p34cdc2. Furthermore, GST-cdc25 dephosphorylated both para-nitrophenyl-phosphate (pNPP) and casein phosphorylated on serine in vitro. Reaction requirements and inhibitor sensitivities were the same as those of phosphotyrosine phosphatases (PTPases). Analysis of cdc25 C-terminal domains from a variety of species revealed a conserved motif having critical residues present at the active site of PTPases. Mutation of the cdc25 Cys480 codon, corresponding to an essential cysteine in the active site of PTPases, abolished the phosphatase activity of GST-cdc25. These data indicate that cdc25 proteins define a novel subclass of eukaryotic PTPases, and strongly argue that cdc25 proteins directly dephosphorylate and activate p34cdc2 kinase to induce M-phase.

Amino Acid Sequence↗
PubMed1991

Two-dimensional gel electrophoresis of human lens epithelium: a study of spatial protein patterns and aging.

The polypeptides in portions of human lens epithelium from individuals of various ages were resolved by two-dimensional (2D) gel electrophoresis. The epithelium was divided into a central area of 12.5 mm2, a surrounding area of 50.2 mm2, and an area which was outside of this region. The proteins were solubilized in urea and run on gels to determine if differences existed in the major polypeptides of the lens epithelium with regard to position in the lens as well as if differences might occur with age. The patterns obtained were remarkably similar in regions of the epithelium examined. The results also demonstrated that the composition of the major polypeptides were not substantially altered in the adult epithelium with age. Two-dimensional gels of proteins from a 3-month-old specimen prepared in a manner similar to that used during extracapsular cataract extraction were also similar to the other samples; however, this sample contained slightly increased staining of proteins greater than 30 kD.

Adult↗
PubMed1991

Effects of naphthalene metabolites on cultured cells from eye lens.

Naphthalene is toxic to the eye and results in opacification of the lens. To investigate the metabolic events that may be occurring in the lens epithelial cells, a cell line of lens from a transgenic mouse was incubated with various metabolites of naphthalene. Naphthoquinone at 50 microM was toxic to most cells with a depletion of glutathione levels noted within 6 h of incubation. At 10 microM, naphthoquinone caused an increase in specific activity of the enzyme DT-diaphorase. This enzyme is thought to be a defense against quinones since semiquinone formation is thought to be lessened. Naphthalene-1,2-dihydrodiol at 50 microM also caused an increase in the specific activity of the DT-diaphorase, while at 10 microM no apparent change occurred in the cells. Although there was evidence of metabolic alterations in the cells with the metabolites of naphthalene, the protein profile by two-dimensional gel electrophoresis did not change and there was no indication of an increase in carbonyl formation in the soluble proteins of the cells. These experiments indicate that the metabolites of naphthalene can cause alteration in the metabolism of the lens cells but may not cause apparent changes in the major proteins within the lens epithelium.

Animals↗
PubMed1991

cdc25 M-phase inducer.

In this paper, we have described the critical experiments leading to the discovery and analysis of the cdc25 M-phase inducer. We have shown that timing of mitosis is sensitive to the level of cdc25+ expression and that the cellular concentration of p80cdc25 increases as cells approach mitosis. From these observations we conclude that, in S. pombe, rate of accumulation of p80cdc25 plays an important role in determining the timing of mitosis. We postulate that under a given set of conditions, a critical level of p80cdc25 activity is required to undergo mitosis. The actual level that is required can vary depending on ploidy, growth rate, nutritional status of the cell, and perhaps other parameters. These signals may be monitored through the weel pathway leading to tyrosyl phosphorylation of p34cdc2. We have shown that p80cdc25 encodes a phosphate that acts by directly dephosphorylating the Tyr-15 residue of p34cdc2. Our studies strongly indicate that this aspect of the mitotic control network is generally conserved among eukaryotes. It is conceivable, however, that the mode of regulation of cdc25 activity may vary from species to species. Clearly, in S. cerevisiae the cdc25+ homolog, MIH1, in contrast to cdc25+, is not rate-limiting for M-phase onset. It will be important to determine whether the level of cdc25+ homologs in other organisms also oscillates during the cell cycle, or whether their activity is controlled by localization or posttranslational mechanisms, such as phosphorylation. Furthermore, our finding of more than one cdc25+ homolog in a single species suggests an additional level of complexity to the control of M-phase onset by cdc25 in higher eukaryotes that will require further investigation.

Amino Acid Sequence↗
PubMed1991

The effects upon helping behaviour of wearing pro-gay identification.

Shoppers were approached either by an ostensibly pro-gay person, wearing a tee-shirt bearing a pro-gay slogan, or by a person wearing an unmarked tee-shirt, asking for change of a pound. Help was less likely to be given to the ostensibly pro-gay person. This effect was not attenuated when the request was justified by a socially acceptable reason for needing change (although, overall, help was more likely to be given when requests were so justified). There were no effects of the sex of the subject or the sex of the person asking for change; nor were there any interactions involving those variables. The results suggest that the negative attitudes towards homosexuals reported in other studies have clear behavioural correlates and that 'homophobia' may be widespread in the general population.

Attitude↗
PubMed1990

Tumour-induced host stromal-cell transformation: induction of mouse spindle-cell fibrosarcoma not mediated by gene transfer.

Tumour-induced host-cell transformation has been addressed by examining human tumours in situ and following xenograft to nude mice. We have found evidence for the transformation of host stromal fibroblasts both in vivo and following the introduction of the tumours to in vitro culture. The in vitro culture of one such xenograft--derived from a human prostatic adenocarcinoma--resulted in the outgrowth of a transformed aneuploid mouse cell line. This transformed line was tumourigenic both in BALB/c nu/nu (nude) mice and in heterozygous nu/+mice, with the morphology of a spindle-cell sarcoma. The cell line did not express human isozymes or human histocompatibility antigens, nor were human chromosomes present. Moreover, human DNA sequences were not detected by human Alu repeat sequence element probing in the transformed cell line grown either in vitro or in vivo. The line contained retroviral long terminal repeat sequences but there was no evidence of proviral activation. These findings indicate that tumour cells may cause transformation of neighbouring stromal cells; that this transformation may proceed in the absence of DNA transfer or activation of endogenous proviruses; and that the means of this observed transformation may involve humoral factors elaborated by the tumour cells.

Adenocarcinoma↗
PubMed1990

Two homologous mixed müllerian tumor lines of the ovary and their characteristics.

Two cell lines, NF and JoN, derived from human ovarian carcinosarcomas, were established in tissue culture and in nude mice. Both lines, growing in monolayers, showed morphologic features of adenocarcinoma cells (NF being aneuploid with a modal number of 53, and JoN being pseudodiploid with a modal number of 44). Intermediate filaments were demonstrated immunohistochemically; the JoN line expressed keratin, but not vimentin or desmin, whereas the NF line expressed vimentin and desmin, but not keratin. Plasminogen activator activity was found in both lines. It is concluded that both of these lines are potentially useful models for studying the diverse characteristics of malignant mixed Müllerian tumors.

Adenocarcinoma↗
PubMed1990

Regulation of mitosis by cyclic accumulation of p80cdc25 mitotic inducer in fission yeast.

The coordination of somatic cell division with cell size must be accomplished by the accumulation of mitotic inducers or the dilution, in the course of cell growth, of mitotic inhibitors. In fission yeast (Schizosaccharomyces pombe), cell size at mitosis is determined by expression of the cdc25+ and nim1+ inducer genes and of the inhibitor gene wee1+, which between them regulate the M-phase protein kinase p34cdc2. We now report that both the phosphoprotein product of cdc25+ (p80cdc25, with apparent relative molecular mass 80,000) and the corresponding messenger RNA increase in concentration as cells proceed through interphase, peaking at mitosis. We propose that the cell-cycle timing of mitosis in somatic cells is regulated by the cyclic accumulation of the cdc25 mitotic inducer, which on reaching a critical level results in activation of p34cdc2 protein kinase. Accumulation of this inducer could play a part in coordinating cell division with growth.

Cell Cycle Proteins↗
PubMed1990

Uterine cervical punch biopsy specimens can be analyzed by 1H MRS.

Biopsy specimens of the uterine cervix, including colposcopically directed punch biopsy specimens of females with atypical Papanicolaou smear tests, are suitable for analysis by magnetic resonance (MR) spectroscopy. A narrow lined lipid MR spectrum, characteristic of malignant tissue, is obtained from a 6-mm3 biopsy specimen of histologically confirmed squamous carcinoma of the cervix. In contrast, specimens containing inflammatory cells generate a broad component only centered at 1.3 ppm with a T2 relaxation value of less than 350 ms. Most biopsy specimens which contain dysplastic cells or evidence of human papilloma virus (HPV) infection have a discernible lipid spectrum similar to that of the malignant tissue specimen. Long T2 relaxation values found in malignant tissue specimens at 1.3 and 1.2 ppm are observed in some but not all of the biopsies which show evidence of HPV infection. The suitability of small tissue samples, such as punch biopsy specimens, for study by MR illustrates the sensitivity of this technique and its potential as an aid to histopathological discrimination between the various precursor states of cervical cancer.

Biopsy↗
PubMed1990

Induction of the enzyme aldose reductase in a lens epithelial cell line from a transgenic mouse.

A lens epithelial cell line established from a transgenic mouse synthesizes high levels of the enzyme aldose reductase which converts sugars to polyols. This enzyme has been implicated in the formation of sugar cataracts in animals and with diabetic complications in man. The mouse aldose reductase has been characterized and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis has an apparent molecular mass of 38,000, similar to the enzyme in rat and man. The cellular enzyme is inhibited by two aldose reductase inhibitors: Sorbinil (IC50 = 1.8 X 10(-7) M) and Alcon 1576 (IC50 = 7.8 X 10(-8) M). The amount and the specific activity of the aldose reductase can be further increased in the cells by raising the osmolarity of the medium to 500 mOSM. Although the amount of aldose reductase is increased approximately sevenfold under these conditions, alpha-crystallin, one of the main lens specific proteins, remained at about the same concentration. No detectable increase in sorbitol was found within the cells, in contrast to published reports on renal cells in which this polyol increases under similar hyperosmotic conditions; however, in the lens cells there was a five-fold increase in the inositol content, suggesting that this polyol rather than sorbitol may be used to compensate for some of the changes in the osmolarity. The induction of the enzyme aldose reductase without the apparent accumulation of its product suggests a complex mechanism for osmoregulation in the lens cells.

Aldehyde Reductase↗
PubMed1990

Interaction of an altered beta-crystallin with other proteins in the Philly mouse lens.

An altered beta B2-crystallin is synthesized in the lens of the Philly mouse. This beta B2 has a more acidic isoelectric point than the beta B2 that is isolated from normal mouse lens. The altered beta B2 is immunologically reactive with antibody to the amino terminal of the beta B2-crystallin, but appears to be present in only very small quantities in the Philly lens. When the soluble proteins are isolated from the Philly lens and chromatographed by gel exclusion chromatography, the beta B2 can be found primarily in the heavy molecular weight fraction. Some immunoreactive material was also found throughout the higher molecular weight beta-crystallin region, beta H, and the lower molecular weight region, beta L. These results would indicate that the altered beta B2-crystallin in the Philly lens can interact with the other beta-crystallins in the lens; however, interactions of the beta B2-crystallin with the other proteins of the lens may cause rapid aggregation of the cellular proteins leading to the formation of the heavy molecular weight material. The increased number of these aggregates may eventually lead to the cataract formation in the Philly mouse.

Animals↗
PubMed1990

Ethanol diffuses across the gastric muscle wall.

Ethanol inhibits the electrical and mechanical activities of gastric smooth muscle, but only at concentrations higher than can be provided by serum delivery. We speculated that the ethanol concentration in the gastric wall may exceed plasma levels by direct diffusion across the mucosa and through the muscle layers. A model of acute ethanol ingestion was created by partitioning the stomachs of three dogs with a surgical stapling device and instilling carbon 14 (14C)-labeled ethanol into the proximal segments. The flux of ethanol was traced by counting 14C in serial sections of the gastric wall. A mucosa-to-serosa gradient of 14C activity was established. The data indicate that direct diffusion from the mucosal surface will produce dramatically higher concentrations of ethanol in gastric muscle than would be anticipated from serum levels alone.

Animals↗
PubMed1990

Human homolog of fission yeast cdc25 mitotic inducer is predominantly expressed in G2.

Entry into mitosis during the somatic cell cycle is regulated in response to signals that monitor the completion of DNA replication, the integrity of the nuclear genome, and, possibly, the increase in cellular mass during the cell cycle. It has been postulated that the operation of this cell cycle control involves the gradual accumulation of rate-limiting mitotic inducers, which trigger nuclear division when their cellular concentration reaches a critical level. We have cloned a human gene, which we call CDC25, whose product may function as a mitotic inducer. This human gene encodes a protein with a predicted molecular mass of 53,000 daltons whose C-terminal domain shares about 37% sequence identity with the fission yeast cdc25+ mitotic inducer. The human CDC25 gene rescues the defect of a fission yeast temperature-sensitive (ts) cdc25ts mutant that is unable to initiate mitosis. In HeLa cells CDC25 mRNA levels are very low in G1 and increase at least 4-fold as cells progress towards M phase. These data suggest that in human cells, as in fission yeast, the accumulation of CDC25 mitotic inducer during G2 may play a key role in regulating the timing of mitosis.

Amino Acid Sequence↗