DROSOPHILA Rae1 FUNCTIONS WITH Hiw/DFsn UBIQUITIN LIGASE COMPLEX TO REGULATE SYNAPTIC DEVELOPMENT IN POST-MITOTIC NEURONS
Document Type
Presentation
Start Date
22-10-2010 3:00 PM
End Date
22-10-2010 4:30 PM
Description
The evolutionarily conserved Hiw/DFsn ubiquitin ligase complex plays essential role in regulating synaptic morphology and function. However, the molecular architecture and action of this ligase complex is not fully understood. Using Tandem Affinity Purification (TAP) techniques, we identified Drosophila Rae1 (DRae1) as a novel component of this complex, which physically interact with both Hiw and DFsn in Drosophila larval and adult brain. Here we show that knocking down DRae1 by RNAi in neurons leads to moderate synaptic overgrowth, and reducing DRae1 gene dose by 50% significantly enhances hypomorphic hiw, but not hiw null, phenotype. Our structure/function analysis maps the Rae1-interacting domain of Hiw into an 1800 aa region in Hiw C-terminus. Interestingly, a Hiw C-terminal transgene that fails to rescue hiw mutants but retains its ability to associate with Rae1 yields a hiw-like synaptic overgrowth phenotype when expressed in neurons, indicating a potent dominant negative effect. This dominant negative effect is completely suppressed by co-expression of DRae1 but not Hiw itself. These data demonstrate that DRae1 functions together with hiw to regulate synaptic morphology. In addition, we show evidence that DRae1 is associates with a mix of RNA species in post-mitotic neurons, and GFPDRae1 transgenic proteins exhibit prominent microtubule localization in axon and synaptic terminal. These observations provide new insights into the molecular mechanisms by which Hiw/DFsn complex may exert its role and/or to be regulated in neuronal tissues.
Recommended Citation
Tian, Xiaolin; DiAntonio, A.; and Wu, C., "DROSOPHILA Rae1 FUNCTIONS WITH Hiw/DFsn UBIQUITIN LIGASE COMPLEX TO REGULATE SYNAPTIC DEVELOPMENT IN POST-MITOTIC NEURONS" (2010). Dr. Joseph M. Moerschbaecher, III Graduate Research Day. 24.
https://digitalscholar.lsuhsc.edu/grad_rs/2010/poster2/24
DROSOPHILA Rae1 FUNCTIONS WITH Hiw/DFsn UBIQUITIN LIGASE COMPLEX TO REGULATE SYNAPTIC DEVELOPMENT IN POST-MITOTIC NEURONS
The evolutionarily conserved Hiw/DFsn ubiquitin ligase complex plays essential role in regulating synaptic morphology and function. However, the molecular architecture and action of this ligase complex is not fully understood. Using Tandem Affinity Purification (TAP) techniques, we identified Drosophila Rae1 (DRae1) as a novel component of this complex, which physically interact with both Hiw and DFsn in Drosophila larval and adult brain. Here we show that knocking down DRae1 by RNAi in neurons leads to moderate synaptic overgrowth, and reducing DRae1 gene dose by 50% significantly enhances hypomorphic hiw, but not hiw null, phenotype. Our structure/function analysis maps the Rae1-interacting domain of Hiw into an 1800 aa region in Hiw C-terminus. Interestingly, a Hiw C-terminal transgene that fails to rescue hiw mutants but retains its ability to associate with Rae1 yields a hiw-like synaptic overgrowth phenotype when expressed in neurons, indicating a potent dominant negative effect. This dominant negative effect is completely suppressed by co-expression of DRae1 but not Hiw itself. These data demonstrate that DRae1 functions together with hiw to regulate synaptic morphology. In addition, we show evidence that DRae1 is associates with a mix of RNA species in post-mitotic neurons, and GFPDRae1 transgenic proteins exhibit prominent microtubule localization in axon and synaptic terminal. These observations provide new insights into the molecular mechanisms by which Hiw/DFsn complex may exert its role and/or to be regulated in neuronal tissues.
Comments
See abstract book page 68