19
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: not found
      • Article: not found

      Thirty-One Flavors of Drosophila Rab Proteins

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Rab proteins are small GTPases that play important roles in transport of vesicle cargo and recruitment, association of motor and other proteins with vesicles, and docking and fusion of vesicles at defined locations. In vertebrates, >75 Rab genes have been identified, some of which have been intensively studied for their roles in endosome and synaptic vesicle trafficking. Recent studies of the functions of certain Rab proteins have revealed specific roles in mediating developmental signal transduction. We have begun a systematic genetic study of the 33 Rab genes in Drosophila. Most of the fly proteins are clearly related to specific vertebrate proteins. We report here the creation of a set of transgenic fly lines that allow spatially and temporally regulated expression of Drosophila Rab proteins. We generated fluorescent protein-tagged wild-type, dominant-negative, and constitutively active forms of 31 Drosophila Rab proteins. We describe Drosophila Rab expression patterns during embryogenesis, the subcellular localization of some Rab proteins, and comparisons of the localization of wild-type, dominant-negative, and constitutively active forms of selected Rab proteins. The high evolutionary conservation and low redundancy of Drosophila Rab proteins make these transgenic lines a useful tool kit for investigating Rab functions in vivo.

          Related collections

          Most cited references55

          • Record: found
          • Abstract: found
          • Article: not found

          DNA cloning using in vitro site-specific recombination.

          As a result of numerous genome sequencing projects, large numbers of candidate open reading frames are being identified, many of which have no known function. Analysis of these genes typically involves the transfer of DNA segments into a variety of vector backgrounds for protein expression and functional analysis. We describe a method called recombinational cloning that uses in vitro site-specific recombination to accomplish the directional cloning of PCR products and the subsequent automatic subcloning of the DNA segment into new vector backbones at high efficiency. Numerous DNA segments can be transferred in parallel into many different vector backgrounds, providing an approach to high-throughput, in-depth functional analysis of genes and rapid optimization of protein expression. The resulting subclones maintain orientation and reading frame register, allowing amino- and carboxy-terminal translation fusions to be generated. In this paper, we outline the concepts of this approach and provide several examples that highlight some of its potential.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Distinct Membrane Domains on Endosomes in the Recycling Pathway Visualized by Multicolor Imaging of Rab4, Rab5, and Rab11

            Two endosome populations involved in recycling of membranes and receptors to the plasma membrane have been described, the early and the recycling endosome. However, this distinction is mainly based on the flow of cargo molecules and the spatial distribution of these membranes within the cell. To get insights into the membrane organization of the recycling pathway, we have studied Rab4, Rab5, and Rab11, three regulatory components of the transport machinery. Following transferrin as cargo molecule and GFP-tagged Rab proteins we could show that cargo moves through distinct domains on endosomes. These domains are occupied by different Rab proteins, revealing compartmentalization within the same continuous membrane. Endosomes are comprised of multiple combinations of Rab4, Rab5, and Rab11 domains that are dynamic but do not significantly intermix over time. Three major populations were observed: one that contains only Rab5, a second with Rab4 and Rab5, and a third containing Rab4 and Rab11. These membrane domains display differential pharmacological sensitivity, reflecting their biochemical and functional diversity. We propose that endosomes are organized as a mosaic of different Rab domains created through the recruitment of specific effector proteins, which cooperatively act to generate a restricted environment on the membrane.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Gal4 in the Drosophila female germline.

              The modular Gal4 system has proven to be an extremely useful tool for conditional gene expression in Drosophila. One limitation has been the inability of the system to work in the female germline. A modified Gal4 system that works throughout oogenesis is presented here. To achieve germline expression, it was critical to change the basal promoter and 3'-UTR in the Gal4-responsive expression vector (generating UASp). Basal promoters and heterologous 3'-UTRs are often considered neutral, but as shown here, can endow qualitative tissue-specificity to a chimeric transcript. The modified Gal4 system was used to investigate the role of the Drosophila FGF homologue branchless, ligand for the FGF receptor breathless, in border cell migration. FGF signaling guides tracheal cell migration in the embryo. However, misexpression of branchless in the ovary had no effect on border cell migration. Thus border cells and tracheal cells appear to be guided differently. Copyright 1998 Elsevier Science Ireland Ltd. All Rights Reserved
                Bookmark

                Author and article information

                Journal
                Genetics
                Genetics
                Genetics Society of America
                0016-6731
                1943-2631
                June 19 2007
                June 2007
                June 2007
                April 03 2007
                : 176
                : 2
                : 1307-1322
                Article
                10.1534/genetics.106.066761
                1894592
                17409086
                37be4b09-1621-4b55-a0e5-1b9582c1e740
                © 2007
                History

                Comments

                Comment on this article