No abstract

The product of the Toll gene is a membrane protein required for the formation of dorso‐ventral polarity during early embryogenesis in Drosophila melanogaster. It acts together with the other dorsal group gene products to specify a nuclear gradient of dorsal morphogen in the syncytial blastoderm stage embryo. Here we report the presence in Toll protein of additional sequences held in common with the human membrane receptor platelet glycoprotein 1b (Gp1b). We propose that these sequences in Toll form disulphide linked extracellular domains that are important for the binding of ligands in the perivitelline space of the embryo. In addition, we show that expression of Toll protein induced in a non‐adhesive cell line promotes cellular adhesion, a property held in common with the related Drosophila glycoprotein chaoptin. Toll protein in such aggregates accumulates at sites of cell‐cell interaction, a characteristic displayed by other cellular adhesion molecules. Taken together these findings suggest that the biochemical function of Toll protein is more closely analogous to that of Gp1b than previously thought.

The blast cells from up to 70% of patients with acute myeloblastic leukaemia exhibit a variable degree of autonomous growth in vitro, which is related to the production of autocrine growth factors. It has recently been established that patients with autonomous blast cell growth have both a lower remission rate and a higher relapse rate, compared to otherwise comparable patients whose blasts exhibit non-autonomous in vitro growth. In a group of 50 patients the actuarial disease-free survival for the autonomous growth group was 11% at 5 years compared to greater than 50% for the non-autonomous growth group. This data suggests that AML blasts with autocrine growth characteristics may be resistant to cytotoxic drug therapy. Here we present further data demonstrating that AML blasts with autonomous growth are relatively resistant to the induction of programmed cell death (apoptosis) and that this is related to the autocrine production of GM-CSF. Also AML blasts with autonomous growths have aberrant expression of genes associated with resistance to apoptosis induced by cytotoxic drugs. These include high expression of the bcl-2 oncoprotein and abnormalities of expression of the p53 tumour suppressor gene. Furthermore bcl-2 expression was found to be unregulated by both exogenous and autocrine GM-CSF suggesting that the documented negative prognostic effect of autonomous growth on treatment outcome in AML, is in part due to the regulatory effect of autocrine GM-CSF on bcl-2 expression, thus protecting cells from apoptosis induced by cytotoxic drug therapy.

The generation of dorso-ventral polarity in Drosophila relies on the formation of a nuclear gradient of the rel/nuclear factor kappa B transcription factor dorsal in the pre-cellular syncitial embryo by a process of differential nuclear localization. It is thought that the gradient is formed by activation at ventral positions of the membrane receptor Toll that in turn causes the local dissociation of dorsal from the cytoplasmic anchor protein cactus. Although Toll is related in its cytoplasmic domain to the interleukin-1 receptor little is known about the signal transduction pathways that lead from Toll to the relocalization of dorsal. In this paper we have used immunofluorescence microscopy as a direct assay of dorsal protein nuclear localization in the Drosophila cell line Schneider 2. We find that increased cytoplasmic calcium concentration and the expression of constitutively active Toll receptors can induce the relocalization of dorsal. By contrast, we find that activation of endogenous protein kinase A and expression of wild-type Toll receptors, which activate zen-chloramphenicol acetyltransferase reporter genes in this system, have only a marginal effect on the cellular distribution of the dorsal protein. Treatment of cells with activators of protein kinase C and radical oxygen intermediates, both of which activate nuclear factor kappa B, also has little effect on dorsal protein localization. We propose that different threshold levels of dorsal activation can be established by distinctly regulated signal transduction pathways.