Biomedical subjects

A Leutz

Publications and source records attributed to A Leutz.

45 records · Page 3Linked to original sources

PubMed1986

v-mil induces autocrine growth and enhanced tumorigenicity in v-myc-transformed avian macrophages.

MH2, an avian retrovirus containing the v-myc and v-mil oncogenes, rapidly transforms chick hematopoietic cells in vitro. The transformed cells belong to the macrophage lineage and proliferate in the absence of exogenous growth factors. Here we analyze a series of MH2 deletion mutants and show that these two oncogenes together establish an autocrine growth system in which v-myc stimulates cell proliferation, while v-mil induces the production of chicken myelomonocytic growth factor (cMGF). We also demonstrate that these two oncogenes cooperate in vivo. MH2 efficiently induces monocytic leukemias and liver tumors, while deletion mutants lacking either a functional v-mil or v-myc do not.

Animals↗

PubMed1984

Purification and characterization of cMGF, a novel chicken myelomonocytic growth factor.

We describe the purification of a novel hematopoietic growth factor from conditioned medium of a transformed macrophage cell line. The factor, termed chicken myelomonocytic growth factor (cMGF) stimulates the growth of chicken myeloblasts transformed by myb oncogene-containing retroviruses and induces the formation of macrophage colonies in uninfected chick bone marrow cultures. The biological activity of the factor is destroyed by trypsin and by reducing reagents but not by SDS. Analysis of crude conditioned medium on non-reducing SDS gels reveals two active species of cMGF with mol. wts. of 23 and 27 kd. Incubation of radioiodinated partially purified cMGF with myeloblasts demonstrates the specific binding of 23- and 27-kd components under non-reducing, and 25- and 29-kd components under reducing conditions. Glycosylation inhibition experiments indicate that the larger molecules represent glycosylated forms of a single protein moiety. The 27-kd species has been purified to homogeneity (80 000-fold enrichment) and exerts its half maximal activity at 2 X 10(-12) M and its maximal activity at 3 X 10(-11) M. Antibodies prepared to purified cMGF completely neutralize the growth-stimulating activity of the factor.

Animals↗

PubMed1984

Autocrine growth induced by src-related oncogenes in transformed chicken myeloid cells.

Chicken myeloid cells transformed by the v-myb-or v-myc-containing leukemia viruses, E26 and OK 10, respectively, require chicken myelomonocytic growth factor (cMGF) for proliferation in vitro. Upon superinfection with retroviruses carrying oncogenes of the src gene family, these myeloid cells acquire the ability to grow in the absence of exogenous cMGF. Conditioned medium prepared from superinfected E26 cells contains a growth-stimulating activity similar in biological and immunological properties to cMGF. This activity is reduced by more than 80% following absorption of conditioned media with antiserum against cMGF. Incubation of superinfected E26 cells with an immunoglobulin fraction of antiserum against cMGF inhibits their proliferation, indicating that the cells are dependent on the secreted factor. We conclude that viral oncogenes of the src family can induce chicken myeloid cells to produce a cMGF-like factor(s) that stimulates proliferation of these cells in an autocrine fashion.

Animals↗

PubMed1984

Ts mutants of E26 leukemia virus allow transformed myeloblasts, but not erythroblasts or fibroblasts, to differentiate at the nonpermissive temperature.

The myb, ets-containing avian acute leukemia virus E26 transforms myeloblasts, erythroblasts, and fibroblasts in culture and causes a mixed erythroid/myeloid leukemia in chicks. We report the isolation and characterization of four E26 mutants that are temperature-sensitive (ts) for myeloblast transformation. At the permissive temperature, tsE26-transformed myeloid cells resemble macrophage precursors and proliferate rapidly, provided the growth medium contains chicken myelomonocytic growth factor (cMGF). When shifted to the nonpermissive temperature the cells stop growing and differentiate into macrophage-like cells, as determined by their expression of morphological, functional, and antigenic markers of normal macrophages. They also lose their responsiveness to cMGF and secrete a cMGF-like factor. Ts mutants of E26 retain their leukemogenicity and their ability to transform both erythroblasts and fibroblasts at the nonpermissive temperature, suggesting that the myb oncogene of E26 causes myeloblast transformation and that ets is responsible for erythroblast and fibroblast transformation.

Animals↗

PubMed1982

Cell type-specificity of epidermal growth factor (EGF) binding in primary cultures of early postnatal mouse cerebellum.

Binding of 125I-labeled epidermal growth factor (EGF) to cells of cultured early postnatal mouse cerebellar cells was investigated by autoradiography in conjunction with cell type-specific immunolabeling of neurons, astrocytes and oligodendrocytes. By use of tetanus toxin for recognition of neurons and glial fibrillary acidic (GFA) protein and 04 antigen as markers for astrocytes and oligodendrocytes, respectively, it could be shown that oligodendrocytes do not express receptors for EGF within the limits of sensitivity of the autoradiographic method, and that less than 1% of all small neurons (mostly granule cells) and 50-90% of all GFA protein-sensitive astrocytes show detectable levels of EGF binding.

Animals↗

PubMed1982

Cultivation of immature astrocytes of mouse cerebellum in a serum-free, hormonally defined medium. Appearance of the mature astrocyte phenotype after addition of serum.

A hormonally defined culture medium is described which supports survival and proliferation of astroglia from primary cultures of early postnatal mouse cerebellum. This medium consists of bovine serum albumin, insulin, transferrin, selenium, hyaluronic acid, protease inhibitor aprotinin, and epidermal growth factor. Trypsin-dissociated single cerebellar cell suspensions are plated in this medium on poly-l-coated glass coverslips and maintained for two weeks before subcultivation. After subcultivation into defined medium more than 99% of all cells are vimentin-positive and fibronectin- and almost completely glial fibrillary acid (GFA) protein-negative, indicating that these cells are less mature astrocytes. After replacement of defined medium by culture medium containing 10% horse serum, expression of GFA protein is detectable in addition to vimentin by indirect immunofluorescence.

Animals↗

PubMed1981

Epidermal growth factor stimulates DNA-synthesis of astrocytes in primary cerebellar cultures.

The capability of epidermal growth factor (EGF) to stimulate DNA-synthesis in neural cells was investigated in primary cultures of early postnatal mouse cerebellum. At concentrations of 10(-8)M, EGF stimulates DNA synthesis in astrocytes, which were identified immunocytologically by the cell type-specific marker, glial fibrillary acidic (GFA) protein. Astrocytes express cell-surface receptors for EGF as can be shown by binding of [125 I]-labeled EGF to live monolayer cultures. In the presence of 10% horse serum, EGF stimulates DNA-synthesis by a factor of about two-fold. Stimulation by EGF over control values is approximately 4-fold in the presence of 1% serum and 6-to 10-fold in the absence of serum. Absolute numbers of astrocytes are increased after more prolonged action of EGF. DNA-synthesis in neurons or oligodendroglia is not significantly stimulated by EGF. EGF enhances cell survival of serum-deprived cerebellar cultures. Fibroblast growth factor does not increase DNA-synthesis in astrocytes under the conditions used in this study.

Animals↗

PubMedDate not supplied

B-Myb, a repressed trans-activating protein.

B-Myb belongs to a family of related transcription factors which share a unique DNA binding domain. B-Myb plays an important role in regulation of the cell cycle. Its expression is upregulated by the human papilloma virus HPV16 E7 oncoprotein. Overexpression of B-Myb can bypass p53-mediated cell cycle arrest. The founding member of the myb gene family, c-Myb, and A-Myb are involved in hematopoiesis and neurogenesis, respectively, and are both activators of gene transcription. Whether B-Myb is a transactivator or a repressor, however, has remained a matter of discussion. We reviewed the transactivation potential of B-Myb in yeast, taking advantage of the fact that inducible gene activation is an evolutionarily conserved process. By mutational analysis we localized a conserved activation domain in B-Myb. In vertebrate cells the transactivation potential of B-Myb is concealed by the C-terminal part of the protein. We show that the cell cycle regulators cyclin A and cyclin E activate B-Myb by eradicating the inhibition mediated by its carboxy-terminus. Our data suggest that in vertebrates the trans-activating function of B-Myb is regulated during the cell cycle and link Myb functions to cell cycle progression.

Amino Acid Sequence↗

PubMedDate not supplied

The c-erb-A protein is a high-affinity receptor for thyroid hormone.

Hormone binding and localization of the c-erb-A protein suggest that it is a receptor for thyroid hormone, a nuclear protein that binds to DNA and activates transcription. In contrast, the product of the viral oncogene v-erb-A is defective in binding the hormone but is still located in the nucleus.

Amino Acid Sequence↗