Protein Targeting and Vesicular Trafficking
Jackson, Mark , Rae, Anne , Casu, Rosanne , Grof, Chris , Bonnett, Graham , Maclean, Don .
A novel targeting motif identified in a sugarcane vacuolar processing enzyme directs green fluorescent proteins into the lytic vacuole.
In the sugarcane stem, the vacuolar compartment occupies a large volume of the storage parenchyma cells. These vacuoles have been regarded as an ideal site for the production and storage of commercially valuable products because of their large size and location in storage tissue. However in previous studies, targeting motifs expected to direct heterologous proteins to the vacuole have not been demonstrated to function efficiently in sugarcane. In the current study we have identified a targeting motif, represented by the sequence IRLPS, present within the amino terminal propeptide of several putative vacuolar proteins from sugarcane. This motif shown to direct an otherwise secreted GFP fusion protein into the large lytic vacuole of both sugarcane callus and mature transgenic sugarcane. In mature sugarcane tissue the stability of GFP appeared to be dependent on cell type, suggesting that the vacuole environment can be hostile to introduced proteins. GFP stability appeared to be enhanced by the addition of concanamycin A, a potent inhibitor of the vacuolar H+ATPase proton pump. This targeting motif is expected to be a valuable tool for engineering sugarcane for production of novel products.
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1 - CSIRO, Plant Industry, QBP, Level 4, 306 Carmody Rd, St Lucia, QLD, 4072, Australia
2 - CSIRO, Plant Industry
3 - University of Queensland, School of Molecular and Microbial Sciences
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM