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Biomedical subjects

N Zhang

Publications and source records attributed to N Zhang.

At least 199 records · Page 11Linked to original sources

Cloning and analysis of expression of a ubiquitin carboxyl terminal hydrolase expressed during oogenesis in Drosophila melanogaster.

Using the enhancer trap approach we have searched for genes with important functions in oogenesis. We selected a line of flies with a P insertion, carrying the Escherichia coli lac Z gene, which showed beta-galactosidase expression in the nurse cell nuclei during oogenesis. Surrounding the P insertion we discovered a cluster of transcription units with enriched expression in the ovary. One of these encodes a protein with extensive sequence similarity to the human and yeast ubiquitin carboxyl terminal hydrolase. Analysis of a fusion protein including the putative ubiquitin carboxyl terminal hydrolase indicated that this protein does have the appropriate enzyme activity, and the gene was assigned the name ubiquitin carboxyl terminal hydrolase uch-D. The expression of this gene is enriched in the nurse cells and transcripts are transported to the embryo. Transcripts are abundant for the first few hours of development. The transcripts are found to be enriched on the ventral side of the oocyte and nurse cells. Rather little is known about the ubiquitin pathway in Drosophila and the discovery of this gene enables us to make predictions as to the roles it may play during early embryogenesis.

Amino Acid Sequence↗

Characterization, sequence, and replication of a small cryptic plasmid from Selenomonas ruminantium subspecies lactilytica.

A 2.5-kb cryptic plasmid, pJDB21, from the gram-negative ruminal anaerobe, Selenomonas ruminantium subspecies lactilytica, was mapped and sequenced. Five open reading frames (ORFs) were predicted and expression of two ORFs was demonstrated. Analysis of the predicted amino acid sequence of the ORF 1 protein indicated approximately 30% homology with the replication protein (rep) common to many gram-positive plasmids, and a highly conserved sequence representing the origin of replication in these plasmids was located upstream of ORF 1. This finding was consistent with a rolling circle form of replication for pJDB21. Transformation of Escherichia coli K-12 UB1636pol Ats with pJDB21 showed that the plasmid replicated independently of DNA polymerase I and produced a single-stranded DNA intermediate. Deletion analyses localized the E. coli replication function to a 1.4-kb sequence that was mapped to the predicted rep gene.

Amino Acid Sequence↗

Sequence and expression of a Drosophila melanogaster cDNA encoding a putative ribosomal protein.

An abundant ovarian cDNA from Drosophila melanogaster has been cloned and sequenced. The predicted protein sequence is similar to that of the ribosomal protein 1024 of Dictyostelium discoideum, the 40S ribosomal protein ys11 of Saccharomyces cerevisiae and a 22 kd protein from Trypanosoma brucei. It seems, therefore, that the Drosophila cDNA also encodes a ribosomal protein. Transcripts are found at all stages of the life cycle but are especially abundant in the ovary, suggesting that this mRNA is maternally stored for utilization in embryogenesis to enable the rapid production of ribosomal proteins and assembly of ribosomes.

Amino Acid Sequence↗

[The influence of prostate-specific antigen p30 on human fertility].

The influence of prostate-specific antigen p30 on human fertility was studied with purified p30, domestic anti-p30 monoclonal and polyclonal antibodies. The results showed that the anti-p30 antibody presented in human serum had no negative effect upon human fertility and the anti-p30 antibody produced by heteroimmunization did not interfere with the process of human fertility in vitro. All these imply that it is unsuitable to recommend p30 as a contraceptive vaccine antigen.

Contraception, Immunologic↗

The HOX-11 (TCL-3) homeobox proto-oncogene encodes a nuclear protein that undergoes cell cycle-dependent regulation.

Molecular analysis of the t(10;14) chromosomal translocation found in pediatric patients with T-cell acute lymphoblastic leukemia has led to the identification of the HOX-11 (TCL-3) protooncogene. The HOX-11 cDNA contains an open reading frame encoding a homeoprotein with features of DNA-binding. The majority of the t(10;14) chromosomal translocation breakpoints have been mapped to the 5' end of the HOX-11 gene, supporting the notion that deregulation of the HOX-11 gene by the t(10;14) chromosomal translocation contributed importantly to leukemia formation. To further define the role of the HOX-11 homeoprotein, we have prepared rabbit antiserum against a trpE-HOX-11 fusion protein. The purified anti-HOX-11 IgG immuno-precipitated a protein with apparent relative molecular mass of 40 kD. Biochemical fractionation demonstrated that the protein is localized in the nucleus. Furthermore, the HOX-11 RNA and protein appeared to be modulated during the cell cycle, with the highest level of expression at G1/S phase boundary. Taken together, these data suggest that the HOX-11 gene product may function as a transcription factor for G1 progression in the cell cycle.

Amino Acid Sequence↗

Migration of Drosophila germ cells: analysis using enhancer trap lines.

Cell migration is a common feature of development. In order to understand more about the factors that control these movements we have embarked on further analysis of the migration of Drosophila germ cells. This process involves passage of the germ cells across the gut primordium and migration toward the mesoderm where the somatic gonad forms. We are particularly interested in the early phase of this migration when the germ cells interact with the amnioproctodeal invagination, the developing gut, before entering into association with the mesoderm. We will summarize the results of our and other studies of these events before describing a number of enhancer trap lines which show expression in the amnioproctodeal invagination during the early phase of germ cell migration. These reveal more about the complexity of this tissue and suggest this tissue is capable of guiding the early phase of germ cell migration.

Animals↗

In search of the identity of the cerebellar climbing fiber transmitter: immunocytochemical studies in rats.

Quantitative immunogold cytochemistry at the electron microscopic level was used to assess the endogenous contents of glutamate, aspartate, homocysteic acid, and glutamine (a precursor of glutamate) in the cerebellar climbing fiber terminals. Of the three excitatory amino acids, only glutamate appeared to be enriched in these terminals. The climbing fiber terminals also displayed immunoreactivity for glutamine. The level of aspartate immunoreactivity was far higher in the nerve cell bodies in the inferior olive than in their terminals in the cerebellar cortex. Homocysteic acid immunolabelling was concentrated in glial cells including the Golgi epithelial cells in the Purkinje cell layer. Our immunocytochemical data indicate that glutamate is a more likely climbing fiber transmitter than aspartate and homocysteic acid.

Amino Acids↗

Detection of anti-SSB antibodies in patients with rheumatic diseases.

Anti-SSB antibodies were measured by ELISA in patients with various kinds of connective tissue diseases using SSB antigen purified from fresh rabbit thymus. The SSB antigen reacted with anti-SSB standard serum, and the positive rates in SS, SLE, RA, PBS and MCTD were 55.1%, 48.3%, 32.8%, 30.8% and 26.3%, respectively. The titers of anti-SSB antibodies were higher in SS and SLE patients than in other connective tissue disease patients. However, 10% of normal individuals were found to have anti-SSB antibodies with low titers. The anti-SSB antibodies detected were mainly of IgG isotype. Preliminary analysis of clinical data showed no relationship between anti-SSB and systemic involvement in SS.

Antibodies, Antinuclear↗

S1 nuclease hypersensitive sites in an oligopurine/oligopyrimidine DNA from the t(10;14) breakpoint cluster region.

Recurring chromosomal translocations are frequently seen in cancers, especially in leukemias and lymphomas. The genes affected by these chromosomal translocations appear to play an important role in oncogenesis. The mechanism underlying the formation of chromosomal translocation is a subject under extensive study. In chromosomal translocations involving the Ig and TCR loci, complete heptamer-spacer-nonamer signal motifs are usually present at the break of the Ig and TCR genes, indicating the involvement of V-D-J recombinase(s). On the other hand, in only about 50% of the cases signal motif sequences have been found at the break in the other participating chromosome, suggesting that different mechanisms may be involved in the scission of the corresponding chromosome. Here we report the identification of an oligopurine/oligopyrimidine DNA in the t(10;14) breakpoint cluster region associated with T-cell acute lymphoblastic leukemia. S1 nuclease mapping revealed multiple S1 hypersensitive sites in the oligopurine/oligopyrimidine DNA. These data suggest a role for oligopurine/oligopyrimidine sequences (non-B DNA) in the formation of chromosomal translocation.

Base Sequence↗

Differential cellular distribution of two sulphur-containing amino acids in rat cerebellum. An immunocytochemical investigation using antisera to taurine and homocysteic acid.

An antiserum to homocysteic acid was raised in rabbits. Immunogens were prepared by coupling this amino acid to bovine serum albumin by means of glutaraldehyde and paraformaldehyde. When applied to semithin or ultrathin sections of rat cerebellum, the antiserum produced selective labelling of glial cells and processes, including the Bergmann fibers. No enrichment of immunoreactivity was detected in nerve terminals of the major excitatory fiber systems. The distribution of homocysteic acid-like immunoreactivity was very different from that of taurine (another sulphur-containing amino acid), as judged from consecutive semithin sections labelled with a postembedding immunoperoxidase procedure and from ultrathin sections labelled with a postembedding double immunogold procedure. Taurine-like immunoreactivity was concentrated in Purkinje cells and was low in glial elements. Our data suggest that the cerebellum contains a glial pool of homocysteic acid (and/or precursors that may undergo spontaneous oxidation to homocysteic acid) and that this amino acid is unlikely to act as a cerebellar transmitter.

Animals↗

Distribution of glutamate-like and glutamine-like immunoreactivities in the rat organ of Corti: a light microscopic and semiquantitative electron microscopic analysis with a note on the localization of aspartate.

The light- and electron microscopic localization of glutamate and glutamine in the rat organ of Corti was studied by means of antisera raised against the respective amino acids coupled to carrier proteins. The light microscopic analysis was performed in semithin sections treated according to the peroxidase-antiperoxidase procedure. The two amino acids were visualized in the same ultrathin sections by use of postembedding immunocytochemistry with two different gold particle sizes. The distribution of aspartate-like immunoreactivity was also recorded, but only at the light microscopic level. In the hair cells, the level of glutamate-like immunoreactivity was higher than that in supporting cells but lower than that in the presumed glutamatergic terminals of cerebellar parallel and mossy fibres. The latter types of terminal were sampled from ultrathin sections that had been incubated under the same conditions as the cochlear sections. Within the hair cells, gold particles signalling glutamate were enriched on mitochondria but not on clusters of synaptic vesicles. Glutamine-like immunoreactivity was present in hair cells as well as supporting cells. The glutamate/glutamine ratio, expressed as the ratio between the respective gold particle densities, was considerably lower for hair cells compared with the cerebellar excitatory terminals. No consistent difference was found between outer and inner hair cells in relation to the levels and subcellular distribution of glutamate and glutamine immunoreactivities. Aspartate-like immunoreactivity was accumulated in outer hair cells, with some labelling also of border cells and Böttcher cells. While the present study confirmed the presence of glutamate in hair cells and demonstrated that these cells are also endowed with the important glutamate precursor glutamine, it revealed notable differences between hair cells and presumed glutamatergic terminals in the CNS. These could reflect differences in the synthesis and compartmentation of transmitter glutamate. Methodological factors could also contribute. Alternatively, the differences could be interpreted to suggest that the hair cell transmitter is not glutamate, but a similar compound. Aspartate could be a candidate in the case of the outer hair cells.

Animals↗

Metabolic compartmentation of glutamate and glutamine: morphological evidence obtained by quantitative immunocytochemistry in rat cerebellum.

An electron microscopic, double-labelling immunocytochemical procedure was used to assess the level of fixed glutamate and glutamine in different cell profiles in ultrathin sections of rat cerebellar cortex. The procedure was based on sequential immunolabelling with two rabbit antisera, using gold particles of different sizes as markers and formaldehyde vapour as a means to avoid interference between the two incubations. Model sections containing a series of known concentrations of the respective amino acids (aldehyde--fixed to rat brain protein) were incubated together with the tissue material. These revealed a close to linear relationship between gold particle density and antigen concentration throughout the range of biological relevance. The ratio between the density of the two categories of gold particles was calculated for the individual profile types. This ratio showed a 20-fold variation, with the highest glutamate/glutamine ratios obtained for putative excitatory terminals (terminals of parallel fibres in the outer part of the molecular layer, followed by mossy and climbing fibre boutons) and the lowest for glial cells (Bergmann glia, astrocytes in the granule cell layer, and oligodendrocytes). Granule cell bodies and dendrites, and cell bodies and processes of putative GABAergic cells (Purkinje, basket and Golgi cells) displayed intermediate ratios. The ratios corresponded to millimolar ratios (mM fixed glutamate/mM fixed glutamine) ranging from 4.5 to 0.2, tentatively assessed by adjusting for differences in labelling efficiency of the two antigens. Our results show that the compartmentation of glutamate and glutamine, an issue previously addressed mainly in the test tube, can be studied in morphologically intact preparations at a resolution that matches the complexity of CNS tissue. The data indicate that glutamate is effectively converted to glutamine in all categories of glial cells, and that glutamate synthesis prevails in each of the three types of excitatory terminals in the cerebellar cortex. Terminals of putative GABAergic cells form a distinct low glutamate/low glutamine compartment.

Animals↗

Synteny mapping in the bovine: genes from human chromosome 5.

In an effort to generate a more complete bovine syntenic map of Type I comparative anchor loci, seven homologs to genes found on HSA5 were mapped using a panel of bovine x rodent hybrid somatic cells. Five HSA5 genes, CSF2, RPS14, PDGFRB, FGFA, and CSF1R, were assigned to bovine syntenic group U22 (chromosome 7), while two others, C9 and HGMCR, mapped to U10 and U5, respectively. Previous studies had assigned the HSA5 marker SPARC to bovine syntenic group U22. The mapping of genes spanning the length of HSA5 in cattle and also in mouse permits syntenic comparisons between prototypic genomes of three mammalian orders, providing insight into the evolutionary history of this region of the ancestral mammalian genome.

Acyl Coenzyme A↗

Synteny mapping in the bovine: genes from human chromosome 4.

Genes homologous to those located on human chromosome 4 (HSA4) were mapped in the bovine to determine regions of syntenic conservation among humans, mice, and cattle. Previous studies have shown that two homologs of genes on HSA4, PGM2 and PEPS, are located in bovine syntenic group U15 (chromosome 6). The homologous mouse genes, Pgm-1 and Pep-7, are on MMU5. Using a panel of bovine x hamster hybrid somatic cells, we have assigned homologs of 11 additional HSA4 loci to their respective bovine syntenic groups. D4S43, D4S10, QDPR, IGJ, ADH2, KIT, and IF were assigned to syntenic group U15. This syntenic arrangement is not conserved in the mouse, where D4s43, D4s10, Qdpr, and Igj are on MMU5 while Adh-2 is on MMU3. IL-2, FGB, FGG, and F11, which also reside on MMU3, were assigned to bovine syntenic group U23. These data suggest that breaks and/or fusions of ancestral chromosomes carrying these genes occurred at different places during the evolution of humans, cattle, and mice.

Animals↗

T-lymphocyte proliferation: tyrosine kinases in interleukin 2 signal transduction.

Interleukin 2 (IL-2)-induced tyrosine phosphorylation appears to play a major role in IL-2-induced cellular proliferation. Several intracellular substrates including the beta chain of the IL-2 receptor complex (IL-2R beta), raf, MAP2 kinase, the regulatory 83 kDa subunit of phosphatidylinositol-3 kinase and S6 kinases are substrates for the IL-2 receptor activated kinase(s). However, none of the identified members of the IL-2 receptor complex exhibits intrinsic tyrosine kinase activity. Therefore, the IL-2R complex must activate intracellular tyrosine kinases. We have demonstrated that specific tyrosine and serine/threonine kinases are coprecipitated with IL-2 receptor constructs that mediate IL-2-induced cell proliferation but not with those that do not. The IL-2-activated tyrosine kinase appears to be associated with a serine and proline rich intracellular domain which is highly conserved between IL-2R beta and the erythropoietin receptor. Although the responsible kinase has not been identified, lck, fyn, fgr, ltk, hck and lyn can be ruled out as obligatory mediators. Using methods to clone tyrosine kinases from T cells, we have identified potential candidate kinases, including several which had not been known to be expressed by T lymphocytes as well as several unique kinases which had not been previously identified in any cell type.

Amino Acid Sequence↗