The highly reactive nature of dopamine renders dopaminergic neurons vulnerable to oxidative damage. We recently demonstrated that loss-of-function mutations in the Drosophila gene Catecholamines up (Catsup) elevate dopamine pools but, paradoxically, also confer resistance to paraquat, an herbicide that induces oxidative stress-mediated toxicity in dopaminergic neurons. We now report a novel association of the membrane protein, Catsup, with GTP cyclohydrolase rate-limiting enzyme for tetrahydrobiopterin (BH4) biosynthesis and tyrosine hydroxylase, rate-limiting enzyme for dopamine biosynthesis, which requires BH4 as a cofactor. Loss-of-function Catsup mutations cause dominant hyperactivation of both enzymes. Elevated dopamine levels in Catsup mutants coincide with several distinct characteristics, including hypermobility, minimal basal levels of 3,4-dihydroxy-phenylacetic acid, an oxidative metabolite of dopamine, and resistance to the vesicular monoamine transporter inhibitor, reserpine, suggesting that excess dopamine is synaptically active and that Catsup functions in the regulation of synaptic vesicle loading and release of dopamine. We conclude that Catsup regulates and links the dopamine synthesis and transport networks.

Cytokines of the TGF-beta superfamily act through an evolutionarily conserved signaling pathway to elicit a diverse range of biological responses in vertebrates as well as invertebrates. Drosophila has proved to be a powerful system to unravel the profound complexities underlying the regulation of this superficially simple signaling system for two reasons--the availability of sophisticated genetic tools and the restricted number of core signaling components compared to vertebrates. A BMP signaling pathway in Drosophila that regulates growth, differentiation and morphogenesis of the embryo and the larva has been extensively characterized. This work has provided major insights into how gradients of secreted proteins can be established and maintained in vivo, allowing a single ligand to induce multiple cell fates rather than function as an on-off switch. More recently, an activin signaling pathway has also been delineated that is required for growth and neuronal function during development. This review provides an overview of TGF-beta signaling in Drosophila with emphasis on the extensive modulation of signaling activity both within and outside the cell, that enables ligands to trigger specific and context-dependent effects.

Catecholamines serve critical neural and developmental roles in Drosophila, as they do in all higher organisms (1–3). The pathway for catecholamine synthesis is shared by Drosophila and mammals, and pathway regulation is evolutionarily conserved. In Drosophila, as in mammals, tyrosine hydroxylase (TH) is the first and rate-limiting enzyme in catecholamine biosynthesis. TH is subject to positive and negative regulation via mechanisms acting at all levels of expression from transcriptional to post-translational. One important source of positive regulation at the post-translational level is the cofactor, 6(R)-5,6,7,8-tetrahydrobiopterin (BH4) (4). The activity of the enzyme absolutely depends upon and is modulated by BH4, the synthesis of which, in turn, is regulated by the activity of the first and rate-limiting enzyme in pteridine biosynthesis, GTP cyclohydrolase I (GTPCH). There are numerous reports of up-regulation of the production and activation of GTPCH and of BH4 synthesis in parallel with or preceding stimulation of catecholamine synthesis (c.f.,5,6). Moreover, ample evidence of the consequences for human catecholamine production when the GTPCH gene is mutated has come to light over recent years (7).

We report the genetic, phenotypic, and biochemical analyses of Catecholamines up (Catsup), a gene that encodes a negative regulator of tyrosine hydroxylase (TH) activity. Mutations within this locus are semidominant lethals of variable penetrance that result in three broad, overlapping effective lethal phases (ELPs), indicating that the Catsup gene product is essential throughout development. Mutants from each ELP exhibit either cuticle defects or catecholamine-related abnormalities, such as melanotic salivary glands or pseudotumors. Additionally, Catsup mutants have significantly elevated TH activity that may arise from a post-translational modification of the enzyme. The hyperactivation of TH in Catsup mutants results in abnormally high levels of catecholamines, which can account for the lethality, visible phenotypes, and female sterility observed in these mutants. We propose that Catsup is a component of a novel system that downregulates TH activity, making Catsup the fourth locus found within the Dopa decarboxylase (Ddc) gene cluster that functions in catecholamine metabolism.

Abstract : We have determined the exon‐intron organization and characterized the 5′‐flanking promoter region of DLG4. Encompassing ~30 kb, the DLG4 locus is composed of 22 exons that range in size from 28 to 1,218 nucleotides. All splice sites conform to the GT‐AG rule, except for the splice acceptor site of intron 5, which is TG instead of AG. Three different exons of DLG4 were found to be alternatively spliced in a subset of tissues. Two of these variants result in altered postsynaptic density 95 (PSD95) isoforms that dramatically truncate the protein. The third splicing variant represents an extension of exon 4 that encodes an additional 33‐amino acid segment. Analysis of the core promoter region for DLG4 suggests that the expression of this gene is controlled by a TATA‐less promoter using a single transcriptional start site embedded within a CpG island. DLG4 maps to a region on chromosome 17p13.1 known to contain a locus for autosomal dominant cone dystrophy 5. Scanning for mutations in the DLG4 coding region and splice sites was performed in 15 cone dystrophy patients, including probands from five families showing linkage to the DLG4 region. No disease‐causing mutations were identified in any patients, suggesting that DLG4 is not the causative gene for this genetic eye disorder.

The MAGUKs (membrane-associated guanylate kinase homologs) are a family of proteins that act as molecular scaffolds for signaling pathway components at the plasma membrane of animal cells. They are localized in and required for the formation of several types of cell junctions, including epithelial tight and septate junctions as well as synaptic and neuromuscular junctions. They are also localized at the plasma membrane of other cell types, including erythrocytes, where they contribute to cell shape maintenance. MAGUKs function mainly by binding directly to the cytoplasmic termini of transmembrane proteins as well as to other signal transduction proteins. They appear to hold together elements of individual signaling pathways, thereby contributing to the efficiency and specificity of signaling interactions while simultaneously maintaining the structural specializations of the plasma membrane. BioEssays 1999;21:912-921.