Mechanism of calcium-independent synaptotagmin binding to target SNAREs

Synaptic vesicle exocytosis requires three SNARE (soluble N-ethylmaleimide-sensitive-factor attachment protein receptor) proteins: syntaxin and SNAP-25 on the plasma membrane (t-SNAREs) and synaptobrevin/VAMP on the synaptic vesicles (v-SNARE). Vesicular synaptotagmin 1 is essential for fast synchronous SNARE-mediated exocytosis and interacts with the SNAREs in brain material. To uncover the step at which synaptotagmin becomes linked to the three SNAREs, we purified all four proteins from brain membranes and analyzed their interactions. Our study reveals that, in the absence of calcium, native synaptotagmin 1 binds the t-SNARE heterodimer, formed from syntaxin and SNAP-25. This interaction is both stoichiometric and of high affinity. Synaptotagmin contains two divergent but conserved C2 domains that can act independently in calcium-triggered phospholipid binding. We now show that both C2 domains are strictly required for the calcium-independent interaction with the t-SNARE heterodimer, indicating that the double C2 domain structure of synaptotagmin may have evolved to acquire a function beyond calcium/phospholipid binding.     

Genomic organization and amplification of the human desmosomal cadherin genes DSC1 and DSC3, encoding desmocollin types 1 and 3

The desmosomal cadherins comprise the desmocollins and desmogleins and are involved in epithelial cell-cell adhesion. There are three desmocollins (DSC 1-3) and three desmogleins (DSG 1-3) that are expressed in a tissue- and development-specific manner. Desmosomal proteins have been implicated in a number of disorders characterized by loss of cell-cell adhesion and trauma-induced skin fragility. Therefore, the desmocollins are potential candidates for genodermatoses involving epithelial tissues. In order to screen the entire DSC1 and DSC3 genes, we have characterized their intron-exon organization. The DSC1 gene comprises 17 exons spanning approximately 33 kb on 18q12.1, and the DSC3 gene comprises 17 exons spanning approximately 49 kb on 18q12.1. We have also developed a comprehensive PCR-based mutation detection strategy for desmocollins 1, 2, and 3 using primers placed on flanking introns followed by direct sequencing of the PCR products.     

Insect Conservation in an Urban Biodiversity Hotspot: The San Francisco Bay Area

The San Francisco Bay Area hosts a diverse insect fauna and a dense cluster of urban areas. The high diversity of insects in the Bay Area arises for three primary reasons: its location in the California biotic province, the diverse local environment and the entomologist-area effect. The juxtaposition of high insect diversity and an area intensively used by humans led to the first recorded extinction as well as the first efforts to conserve insects in the United States. Habitat loss due to urbanization, agriculture, and invasive species is largely responsible for local extinctions and reduction in abundance of the remaining species. Invasive species such as the Argentine ant and pathogens causing mortality of oaks and pines are poised to have substantial impacts on the insect fauna of the Bay Area in the near future. Understanding which taxa can or cannot persist in remnant habitat patches within an urban or agricultural matrix, and what management practices would encourage persistence should be a focus of future research. Assessments of population status should be focused on insects at risk of extinction because of their restricted geographic ranges, low vagility, interactions with invasive species, or known reduction in their habitat. Assessments that combine examinations of museum collections, literature, and field surveys might enable determination of the status of many species within the Bay Area. Such an approach might better define the scope and magnitude of the problem of conserving insects in an increasingly urbanized region.