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E Barklis

Publications and source records attributed to E Barklis.

39 records · Page 3Linked to original sources

Specific cell-cell contacts are essential for induction of gene expression during differentiation of Dictyostelium discoideum.

Postaggregation Dictyostelium discoideum cells contain 2000-3000 mRNA species that are absent from pre-aggregation cells. These aggregation-dependent sequences compose 30% of the mass of the late mRNA and represent the transcription products of an additional 11% of the single-copy genome. By analysis of mutants that are blocked at different stages of differentiation, we show that induction of expression of these genes is correlated with the formation of tight cell-cell contacts that resist EDTA. In particular, mutants that exhibit chemotaxis and aggregate to form loose mounds but do not form cell-cell contacts that resist EDTA fail to induce these late mRNA and protein species. By contrast, mutants that form normal contacts but progress no further through development do express the late mRNA species. Thus, interactions at the cell surface are involved in developmental induction of a large group of coregulated mRNAs. We have employed two independent assays for these developmentally regulated mRNAs: hybridization of gel-separated RNAs to cloned nuclear DNAs and hybridization of mRNA to a cDNA probe specific for the population of 2000-3000 regulated sequences.

Journal Article↗

S-180 cells secrete nerve growth factor protein similar to 7S-nerve growth factor.

The nerve growth factor protein (NGF) has been identified by biological assay and rocket immunoelectrophoresis in media conditioned by monolayers of mouse S-180 sarcoma cells, a transformed cell line of salivary gland origin. By utilization of a purification procedure designed to enrich for acid-dissociable, high-molecular-weight complexes similar to the 7S-NGF isolated from male mouse submaxillary gland, it has been determined that a significant portion of mouse sarcoma NGF is initially present as a complex at neutral pH with a molecular weight indistinguishable from that of 7S-NGF. At pH 4.0 the mouse sarcoma NGF complex dissociates, and the active subunit can be isolated as a species with a molecular weight of less than 40,000. The induced dissociation at pH 4.0 of the mouse sarcoma NGF high-molecular-weight complex, as well as the complex's behavior in isoelectric focusing and sucrose gradient sedimentation is consistent with the hypothesis that 7S-NGF is packaged as a subunit containing protein intracellularly prior to secretion into the extracellular space. Moreover, the stability of the mouse sarcoma NGF complex at dilute concentrations is similar to that reported for the purified 7S-NGF complex.

Animals↗

Mechanism of sequential induction of cell-type specific mRNAs in Dictyostelium differentiation.

Upon starvation, the cellular slime mould Dictyostelium discoideum initiates a 24-h programme of differentiation. Within 6 h, cells move towards aggregation centres in response to pulsatile synthesis and secretion of cyclic AMP. At about 12 h, aggregates of 10(5) cells are formed, held together by newly made surface adhesion molecules. The cells then differentiate into the two principal types found in the terminal stage of development, spores and stalks. Here we show that the chemotaxis and aggregation stages of this developmental programme can be described as a series of sequential events in which these extracellular signals--starvation, cyclic AMP and cell-cell contact--induce specific, sequential changes in the pattern of gene expression.

Cell Differentiation↗