Biomedical subjects

A Seddon

Publications and source records attributed to A Seddon.

9 recordsLinked to original sources

PubMed1992

Anti-viral activity of human recombinant heparin-binding proteins HBNF and MK.

Herpes simplex viruses bind to cell surface heparan sulfate proteoglycans, as a first step of viral infection. We report here that two recombinant heparin-binding proteins HBNF and MK inhibit infectivity of human herpes simplex viruses types 1 and 2 and human cytomegalovirus. Carboxymethylated HBNF and MK, which retain affinity for heparin-Sepharose, do not exhibit anti-viral activities. Arguments are presented that anti-viral effects of HBNF and MK are due to the competition for the specific binding to the cell surface heparan sulfate proteoglycans.

Alkylation↗

PubMed1991

Structure/activity relationships in basic FGF.

Although the FGFs have been subject to extensive biological studies, only limited progress has been made so far in determining the critical elements of structure-activity relationships in the FGFs. Among the recognized structural elements with potential to affect the biological activity of FGFs are the cysteine residues, and the heparin- and receptor-binding domains. These features have been studied using a variety of experimental approaches, but the available data are inconclusive. For example, ambiguity regarding the presence of a disulfide structure in FGFs was not resolved until the availability of x-ray crystal structure data. Furthermore, the functionally important heparin- and receptor-binding domains have been poorly characterized, with some interpretations being controversial. In this report, we describe a novel fragment of basic FGF (bFGF) with high biological activity [Ser78,96-bFGF(70-153)]. This fragment was generated by pronase treatment of heparin-bound recombinant Glu3,5Ser78,96-bFGF mutant and is active in vitro at an ED50 of about 100 ng/ml. The structure of the fragment and the manner by which it was generated provide additional insight into important aspects of structure-activity relationships in FGFs. Specifically, we conclude that (a) the cysteines in our bFGF mutant do not form a disulfide bond, (b) the high-affinity heparin binding of bFGF critically depends on an intact 3-dimensional structure of the growth factor rather than on specific heparin-binding sequence domains, and (c) the bFGF sequence between residues 70 and 122 is important for high biological activity.

Binding Sites↗

PubMed1991

Isolation from bovine brain and structural characterization of HBNF, a heparin-binding neurotrophic factor.

A heparin-binding protein with neurotrophic activity for perinatal rat neurons, termed HBNF, was purified to homogeneity from bovine brain utilizing pH 4.5 extraction, ammonium sulfate precipitation, cation exchange and heparin-Sepharose affinity chromatographies, and reverse phase HPLC. In the presence of protease inhibitors during extraction, a protein with an apparent molecular weight of 18 kDa was obtained in a yield of approximately 0.5 mg/kg brain tissue. The amino acid sequence of the first 114 residues of HBNF was determined and found to highly homologous to the cDNA-derived amino acid sequence of human HBNF, a 136-residue protein. Bovine and human HBNFs have identical molecular weights as judged by SDS gel electrophoresis and very similar amino acid compositions. This and overall sequence conservation suggest that bovine HBNF is also a 136 amino acid protein with a calculated molecular weight of approximately 15.5 kDa. The apparent discrepancy between calculated and observed molecular weights of bovine HBNF (and of human HBNF of which the complete sequence is known) is most likely a result of the highly basic nature of HBNF. If protease inhibitors were omitted during tissue extraction, two additional proteins with lower apparent molecular weights and identical N-terminal sequences were isolated, with the smallest forms being the major product. Amino acid analysis showed that the smaller forms correspond to C-terminally truncated HBNFs with calculated molecular weights of 13.6 and 12.4 kDa, lacking approximately 14 and 22 residues. Comparison of the HBNF protein sequence with sequences stored in the Protein Identification Resource/Genbank databases reveals high homology to the translation product of the MK-1 gene, which is retinoic acid-inducible in embryonic carcinoma cells and developmentally expressed during gestation in mice.

Amino Acid Sequence↗

PubMed1990

The identification of a second cell cycle control on the HO promoter in yeast: cell cycle regulation of SW15 nuclear entry.

HO encodes a site-specific endonuclease that initiates mating type switching in S. cerevisiae. It is expressed only transiently during the cell cycle of mother cells, as they undergo Start, but not in daughter cells. Since SWI5 appears to be the only HO transcription factor missing when daughter cells undergo Start, we were interested in the intracellular distribution of SWI5 at cell division. We discovered that SWI5 is found equally concentrated in the nuclei of both mother and daughter cells at the end of anaphase, suggesting that its subsequent fate must somehow differ. Prior to the end of anaphase, SWI5 accumulates in the cytoplasm and only moves into the nucleus when cells enter G1. A version of the HO promoter that has lost its dependence on Start is nevertheless still strongly cell cycle regulated and is activated when SWI5 moves into the nucleus.

Cell Compartmentation↗

PubMed1989

Vascular endothelial cell function and ultrastructure in thrombotic microangiopathy following allogeneic bone marrow transplantation.

We report studies on vascular endothelial function and ultrastructure in 2 cases of fatal cyclosporin (CS)-associated thrombotic microangiography following allogeneic bone marrow transplantation (BMT). Spontaneous vascular release of prostacyclin (PGI2) from a vein sample ex vivo was absent, and scanning electron microscopy (SEM) showed surface changes indicative of vascular endothelial damage (case 1). PGI2 release from cultured human umbilical vein endothelial cells incubated with patients' serum in vitro was normal in both cases. Plasma von Willebrand factor (vWF) antigen and ristocetin cofactor activity levels were raised in both patients, 5.06 and 7.02 (case 1) and 3.60 and 2.01 (case 2) (normal ranges 0.59-1.57 and 0.42-1.74 U/ml), respectively, but multimer patterns were normal. The SEM appearances coupled with the absent PGI2 release and raised vWF levels suggest that vascular endothelial damage is central to the pathogenic process in thrombotic microangiopathy following allogeneic BMT but the mechanisms appear to be distinct from those in the haemolytic uraemic syndrome and de novo thrombotic thrombocytopenic purpura. The precise role of CS in this process remains to be identified.

Adult↗

PubMed1988

Characterization of a transcription factor involved in mother cell specific transcription of the yeast HO gene.

The yeast HO gene, which encodes an endonuclease involved in initiating mating type interconversion, is expressed in mother cells but not in daughters. It has been demonstrated that the SWI5 gene, which is an activator of HO expression, plays a critical role in this differential mother/daughter expression of HO. In this paper we describe the cloning and sequencing of the SWI5 gene. The predicted amino acid sequence derived from the cloned SWI5 gene shows homology with the repeated DNA-binding domains ('zinc fingers') of Xenopus transcription factor TFIIIA. A region of the HO promoter involved in the SWI5-dependent transcriptional activation of HO was identified by deletion analysis of the HO promoter in the chromosome, and by testing the ability of HO DNA fragments to activate transcription in the context of a heterologous promoter. The SWI5 gene product was overproduced in yeast from the GAL1-10 promoter, since the SWI5 protein is made at very low levels in wild-type strains, and protein extracts were used to demonstrate that the SWI5 protein binds in vitro to a segment of the HO promoter required for transcriptional activation in vivo.

Amino Acid Sequence↗

PubMed1987

Cell cycle regulation of SW15 is required for mother-cell-specific HO transcription in yeast.

In haploid homothallic yeast, cell division gives rise to a mother cell that transiently transcribes the HO gene (as it undergoes START) and a daughter cell that does not. Consequently, only mother cells switch their mating types. Here, we test the proposition that a transcription factor called SWI5 is the "determinant" of mother-cell-specific HO transcription; that is, that SWI5 is the only factor missing in daughter cells. We show that SWI5 RNAs are cell-cycle regulated so that they are only produced after the post-START window of HO transcription. This regulation is vital for mother-cell specificity since constitutive transcription of SWI5 causes daughter cells to switch their mating types. We propose that SWI5 gene products are partitioned asymmetrically at cell division.

Base Sequence↗

PubMed1987

Stimulation of fetal growth associated with decreased maternal plasma oestrogen levels.

Reduced maternal and fetal plasma oestrogen levels were produced in rats by daily administration of an LH-RH agonist (obtained from IMPERIAL CHEMICAL INDUSTRIES; ICI product 118630) to the dams from day 12 of gestation. Fetuses from the treated mothers were significantly heavier and longer at delivery on day 21 of gestation than fetuses from control mothers. Placental weights were also greater in the treated group.

Animals↗

PubMed1986

Immuno-neutralization of maternal plasma oestradiol stimulates fetal growth in rats.

On day 12 of pregnancy, 7 rats were injected with 2 ml of an anti-oestradiol antiserum a quantity capable of binding 2.4 micrograms oestradiol. Seven further (control) rats were injected with an antiserum raised against human serum globulin. After sacrifice on day 21 of pregnancy, the fetuses from treated rats showed an increase in both placental weight (0.851 +/- 0.023 g, n = 98 vs 0.752 +/- 0.019 g, n = 97) and fetal weight (4.446 +/- 0.426 g, n = 98 vs 3.403 +/- 0.211 g, n = 97). There was no significant difference in protein, DNA and RNA contents between the fetuses from treated mothers and those from control mothers. This experiment confirms that oestrogens may play a role in fetal weight gain.

Animals↗