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N K Moschonas

Publications and source records attributed to N K Moschonas.

31 records · Page 2Linked to original sources

Molecular cloning, structure and expression analysis of a full-length mouse brain glutamate dehydrogenase cDNA.

We isolated and analysed a full-length mouse brain glutamate dehydrogenase (GLUD) cDNA as a preliminary step to use the mouse model for the investigation of GLUD function in neurotransmission and neurodegeneration. GLUD coding sequences were found highly conserved among mouse, human and rat. Northern blots revealed two transcripts with different ratios in different mouse organs implying some mechanism of tissue-specific expression. In contrast to human, mouse GLUD gene family appears not to contain an intronless member.

Amino Acid Sequence↗

Chromosomal mapping of glutamate dehydrogenase gene sequences to mouse chromosomes 7 and 14.

Glutamate dehydrogenase (GLUD) plays an important role in mammalian neuronal transmission. In human, GLUD is encoded by a small gene family. To determine whether defects in Glud genes are associated with known neurological mutations in the mouse and to contribute to the comparative mapping of homologous genes in man and mouse, the chromosomal location of genes reactive with a mouse brain GLUD cDNA were determined. Genomic Southern analysis of a well-characterized panel of Chinese hamster x mouse somatic cell hybrids identified two GLUD-reactive loci, one residing on mouse Chromosome 14 and the other on Chromosome 7. Progeny of an intersubspecies backcross were used to map one of these genes, Glud, proximal to Np-1 on Chromosome 14, but no restriction fragment polymorphisms could be identified for the second locus, Glud-2.

Animals↗

Developmental and inducible patterns of human theta 1-globin gene expression in embryonic/fetal and adult erythroid cells.

Human theta (theta 1)-globin gene represents a member of the alpha-like globin gene family residing on chromosome 16. theta 1-Specific transcripts have been detected so far only in erythroid tissues and in erythroleukemia K562 cells. To investigate systematically its inducible expression and developmental specificity, we analyzed at the RNA level five additional human erythroleukemia cell lines with diverse developmental globin programs, two somatic cell hybrids between K562 and mouse erythroleukemia (MEL) cells, a human fetal liver x MEL somatic cell hybrid, and reticulocytes and bone marrow cells from normal adults. theta 1-Globin gene was expressed in all cell types. Inducible expression (two- to sixfold) was documented both in HEL and K562 erythroleukemia cells after 5-azacytidine treatment. Like K562 cells, HEL cells also displayed hemin-inducible theta 1-globin gene expression. Following transfer of human chromosome 16 from embryonic/fetal K562 to the adult MEL cells, theta 1-globin gene remained active but lost its potential for inducibility, suggesting probably a trans regulation mechanism. Higher levels of theta 1 mRNA were found in fetal liver cells compared with trace amounts in reticulocytes and normal adult bone marrow cells. These data clearly show that in contrast to the embryonic and adult patterns of expression of zeta and alpha-globin genes, respectively, theta 1-globin gene displays a different profile, being active predominantly during the early stages of ontogeny, switching to lower levels of expression in adulthood.

Embryonic and Fetal Development↗

Locus assignment of human alpha-globin structural mutants by selective enzymatic amplification of alpha 1 and alpha 2-globin cDNAs.

We have used the powerful methodology of DNA enzymatic amplification in order to assign human alpha-globin structural mutants to one of the two highly homologous alpha-globin genes. Selectively amplified alpha 1 and alpha 2-globin cDNAs were dot-blotted and further hybridized to synthetic oligonucleotides encompassing either the normal or the mutated sequences. The generated signals corresponded specifically to one of the two alpha-globin genes. Using this approach the alpha-globin structural mutants J-Buda and G-Pest were found to be encoded by the alpha 2 and the alpha 1-globin genes, respectively. Furthermore, the exact nucleotide changes were determined. We propose this technique to serve as a simple and definitive method for assigning alpha-globin structural mutants.

Base Sequence↗

Evolution of chorion structural genes and regulatory mechanisms in two wild silkmoths: a preliminary analysis.

We report a preliminary analysis of structural and regulatory evolution of the A and B chorion gene families in two wild silkmoths, Antheraea pernyi and Antheraea polyphemus. Homospecific and heterospecific dot hybridizations were performed between previously characterized A. polyphemus complementary DNA clones and total or stage-specific follicular mRNAs from the two species. The hybridization patterns indicated substantial interspecies changes in the abundance of corresponding mRNA sequences (heteroposic evolution) without substantial changes in their developmental specificities (heterochronic evolution). In addition, the proteins encoded in the two species by corresponding mRNAs were determined by hybrid-selected translation followed by electrophoretic analysis. The results suggested that the proteins evolve in size, presumably through internal deletions and duplications.

Animals↗

The B multigene family of chorion proteins in saturniid silkmoths.

The main features of the B family of chorion proteins in saturniid moths were examined by partial sequencing of representative B proteins, seven from Antheraea polyphemus and two from A. pernyi. Comparisons were made to sequences derived from seven recombinant DNA clones representing three types of B family proteins of A. polyphemus. The central regions of the sequences are conservative, both within and between moth species, and differ largely by a few amino acid replacements, rather than deletions or insertions. By contrast, the amino-terminal third varies more substantially, in a manner which defines two protein subfamilies: within each subfamily sequences are similar, but the subfamilies differ by at least two multiresidue deletions as well as by amino acid replacements. These properties are analogous to features of the A family of chorion proteins.

Amino Acid Sequence↗

Evolution of a multigene family of chorion proteins in silkmoths.

The evolution of the A family of chorion genes was examined by comparing new protein and DNA sequences from the silkmoths Antheraea pernyi and Bombyx mori with previously known sequences from Antheraea polyphemus. The comparisons indicated that the A family and its major subfamilies are ancient and revealed how parts of the genes corresponding to distinct regions of the protein structure have evolved, both by base substitutions and by segmental reduplications and deletions.

Amino Acid Sequence↗

Chromosomal mapping of two members of the human glutamate dehydrogenase (GLUD) gene family to chromosomes 10q22.3-q23 and Xq22-q23.

Glutamate dehydrogenase (GLUD) is an important mitochondrial enzyme that participates in neuronal transmission by catalyzing the deamination of L-glutamate, which serves as a potent excitatory neurotransmitter. The direct involvement of GLUD in the pathogenesis of certain human neurodegenerative disorders has been suggested recently. To investigate its possible role in the induction and progression of these disorders, we have initiated studies focusing on the chromosomal organization of the several members of the GLUD family and their functional status. In the present study using a panel of human x rodent somatic cell hybrids and in situ hybridization to metaphase chromosomes, we documented that the members of the GLUD gene family are dispersed in the human genome. The functional GLUD1 gene was mapped to chromosome 10q22.3-q23, and an intronless processed gene (GLUDP1) to chromosome Xq22-q23, while the truncated intron-containing GLUD pseudogene GLUDP2 was also assigned on chromosome 10, but not closely linked to the GLUD1 gene. These results provide novel information concerning the chromosomal organization of the human GLUD gene family.

Animals↗

[A normalized cDNA library from human erythroleukemia cells].

Normalized cDNA library from human erythroleukemia cells has been constructed. For equalizing frequencies of different cDNAs denaturation was carried out followed by partial reassociation. Single-stranded cDNAs separated by hydroxyapatite chromatography were transformed into double-stranded form by PCR and cloned in lambda gt11. Frequencies of 10 control nucleotide sequences were estimated. After normalization frequencies of the abundant sequences decreased. The normalized cDNA library may be used for search of the clones corresponding to the rare mRNAs and for human genome mapping.

Base Sequence↗