Jupiter, a newDrosophila protein associated with microtubules

Abstract

In this study we describe a novel Drosophila protein Jupiter, which shares properties with several structural microtubule-associated proteins (MAPs) including TAU, MAP2, MAP4. Jupiter is a soluble unfolded molecule with the high net positive charge, rich in Glycine. It possesses two degenerated repeats around the sequence PPGG, separated by a Serine-rich region. Jupiter associates with microtubules in vitro and, fused with the green fluorescent protein (GFP), is an excellent marker to follow microtubule dynamics in vivo. In a jupiter transgenic Drosophila strain generated by the "protein-trap" technique, Jupiter:GFP fusion protein localizes to the microtubule network through the cell cycle at the different stages of development. We found particularly high Jupiter:GFP concentrations in the young embryo, larval nervous system, precursors of eye photoreceptors and adult ovary. Moreover, from jupiter:gfp embryos we have established two permanent cell lines presenting strongly fluorescent microtubules during the whole cell cycle. In these cells, the distribution of the Jupiter:GFP fusion protein reproduces microtubule behavior upon treatment by the drugs colchicine and taxol. The jupiter cell lines and fly strain should be of wide interest for biologists interested in in vivo analysis of microtubule dynamics.

Reliance of Wolbachia on High Rates of Host Proteolysis Revealed by a Genome-Wide RNAi Screen of Drosophila Cells

“…Robotics and automated microscopy were used to fix, stain, and image the cells (see Materials and Methods). The JW18 cell line expresses a GFP-Jupiter fusion protein that labels the cytoskeleton (Karpova et al 2006), thus highlighting the entire cytoplasm and indicating cell boundaries. Anti-histone staining labeled the host nuclei, and DAPI was used to stain both host nuclei and cytoplasmic Wolbachia.…”

“…The JW18 cell line bearing the Jupiter-GFP transgene was generated according to previously described methods (Karpova et al 2006;Serbus et al 2012;Debec et al 2016). Two-to 15-hour-old embryos derived from Wolbachia-infected flies carrying a Jupiter-GFP transgene were homogenized and plated in flasks.…”

“…Wolbachia is stably maintained in these cultures and exhibits a close association with microtubules as found in Drosophila somatic and germline tissues (Kose and Karr 1995;Albertson et al 2009). The cell line expresses a transgene encoding the GFP-tagged gene Jupiter, which encodes a microtubule-associated protein that labels microtubules and facilitates high-throughput, cell-based screening approaches (Karpova et al 2006). The Wolbachia-infected cells were originally employed for high-throughput, cell-based screens to identify small molecule inhibitors of Wolbachia (Serbus et al 2012;Van Voorhis et al 2016).…”

GFP markers for studying D. melanogaster spermatogenesis

“…One of the sets of such protein traps was obtained in Villefranche (France) by the team headed by A. Debec. Several chimeric proteins from this set have been characterized [2][3][4].…”

In vivo analysis of MT‐based vesicle transport by confocal reflection microscopy

“…To label the MTs, a Jupiter-GFP expressing protein trap line was used, a kind gift from Alain Debec. [Jupiter-GFP, -Jup-GFP for short -is a MT associated protein with thus far unknown function; Karpova et al, 2006]. The Jup-GFP provides better contrast MT images than any of the conventionally used Tau-GFP [Murray et al, 1998] or the Tubulin-GFP lines [Grieder et al, 2000] and does not effect viability and fertility perceptibly (data not shown).…”