Metabolic flux analysis of CHO cells at growth and non-growth phases using isotopic tracers and mass spectrometry.

Transforming growth factors (TGF) are defined as biologically active polypeptides which reversibly confer the transformed phenotype onto untransformed cultured cells. They have been subdivided into two classes: type α and type β TGFs. TGF-β acts synergistically with TGF-α in inducing phenotypic transformation. TGF-β can also act as negative autocrine growth factor. A human 1050-bp EcoRI cDNA fragment was used to map the human locus for TGF-β by Southern blotting of DNA prepared from 17 human × Chinese hamster somatic cell hybrids. The humanspecific restriction fragments segregated with human chromosome 19 in all of 14 informative hybrids. All other human chromosomes were discordant with the TGF-β bands in at least four hybrids. After in situ hybridization of the tritiated TGF-β probe to normal human metaphase spreads, 151 silver grains were scored in 54 cells. Of 24 grains over chromosome 19, 16 grains (11%) lay over region 19q13.1 → q13.3. Of the 54 cells analyzed, 16 (30%) had label over region 19q13.1 → q13.3. Thus,TGFB is assigned to chromosome 19, subbands q13.1 → q13.3. TheTgf- β locus in the mouse was mapped to chromosome 7 by hybridizing a murine cDNA probe to a Chinese hamster × mouse hybrid panel. Human chromosome 19 and proximal mouse chromosome 7 share another four homologous loci.

Transforming growth factor β gene maps to human chromosome 19 long arm and to mouse chromosome 7

Because others had described a lack of the enzyme adenosine deaminase as associated with severe combined immunodeficiency, we surveyed kindreds with infants affected with such an immunodeficiency. Three infants in two families with severe combined immunodeficiency were found to have no detectable erythrocyte adenosine deaminase. Eleven family members heterozygous for adenosine deaminase deficiency were encountered among the first-degree relatives; adenosine deaminase deficiency and severe combined immunodeficiency were associated and inherited as autosomal recessive traits in both kindreds. Successful bone-marrow transplantation was carried out in two of these infants. Normal immunologic function was established in both children, but the deficiency of adenosine deaminase persisted in their erythrocytes. The enzyme deficiency did not impair the successful establishment of normal humoral and cellular immunity by transplants of bone-marrow cells from siblings who were either normal or heterozygous for adenosine deaminase deficiency.

Severe combined immunodeficiency and adenosine deaminase deficiency.

Human probes identifying the cellular homologs of the v-ets gene, Hu-ets-1 and Hu-ets-2, and two panels of rodent-human cell hybrids were used to study specific translocations occurring in acute leukemias. The human ets-1 gene was found to translocate from chromosome 11 to 4 in the t(4;11)(q21;23), a translocation characteristic of a subtype of leukemia that represents the expansion of a myeloid/lymphoid precursor cell. Similarly, the human ets-2 gene was found to translocate from chromosome 21 to chromosome 8 in the t(8;21)(q22;q22), a nonrandom translocation commonly found in patients with acute myeloid leukemia with morphology M2 (AML-M2). Both translocations are associated with expression different from the expression in normal lymphoid cells of ets genes, raising the possibility that these genes play a role in the pathogenesis of these leukemias.

http://science.sciencemag.org/content/sci/231/4736/379.full.pdf

Hu-ets-1 and Hu-ets-2 genes are transposed in acute leukemias with (4;11) and 8;21) translocations

Techniques are described for the electrophoretic characterization on Cellogel of adenosine deaminase, adenylate kinase, enolase, fumarase, α-galactosidase, glucosephosphate isomerase, glutathione reductase, glutamic-oxaloacetic transaminase, guanylate kinase, hexosaminidase, hypoxanthine-guanine phosphoribosyltransferase, indophenol oxidase, “malic” enzyme, mannosephosphate isomerase, nucleoside phosphorylase, peptidase-A, peptidase-B, peptidase-C, phosphoglucomutase, pyruvate kinase, triosephosphate isomerase, and UDP glucose pyrophosphorylase1 in the man-Chinese hamster somatic cell hybrid systems.

Enzyme electrophoresis on cellulose acetate gel. ii. zymogram patterns in man-chinese hamster somatic cell hybrids

About 75 man-Chinese hamster hybrid clones were analysed for their human chromosome complement and simultaneously tested for human enzyme markers. Correlation of the presence of chromosomes and enzyme activity revealed assignments of the PGD linkage group to chromosome 1, ME1, PGM3 and IPO-B to 6, LDH-A to 11, LDH-B to 12 and IPO-A to 21.

Localization of genes on human chromosomes by studies of human-Chinese hamster somatic cell hybrids. Assignment of PGM3 to chromosome C6 and regional mapping of the PGD, PGM1 and pep-C genes on chromosome A1.

A human-Chinese hamster hybrid cell line was treated with X-rays and subsequently cloned. In two clones, segregation between three markers assigned to the human A1 chromosome (6-phosphogluconate dehydrogenase, phosphoglucomutase 1 and peptidase C) was found. Chromosome analysis revealed an aberrated human A1 chromosome in both clones. The results suggest the following gene sequence on chromosome A1: 6 PGD, PGM1 (on the short arm), Pep C (on the long arm).

Cytological mapping of the genes assigned to the human A 1 chromosome by use of radiation-induced chromosome breakage in a human-Chinese hamster hybrid cell line.

The presence or absence of human hexosaminidases A and B was studied in 105 man-Chinese hamster hybrid cell clones. The results show that these two isoenzymes behave as independent markers.

Harry van Someren

Papers

Enzyme electrophoresis on cellulose acetate gel. ii. zymogram patterns in man-chinese hamster somatic cell hybrids

Localization of genes on human chromosomes by studies of human-Chinese hamster somatic cell hybrids. Assignment of PGM3 to chromosome C6 and regional mapping of the PGD, PGM1 and pep-C genes on chromosome A1.

Cytological mapping of the genes assigned to the human A 1 chromosome by use of radiation-induced chromosome breakage in a human-Chinese hamster hybrid cell line.

Independent loss of human hexosaminidases A and B in man-Chinese hamster somatic cell hybrids

Mammalian cell culture studies