Richard Scheller - Academia.edu (original) (raw)

Uploads

Papers by Richard Scheller

The Journal of cell …, 1992

Potential interactions between membrane components of rat brain synaptic vesicles were analyzed b... more Potential interactions between membrane components of rat brain synaptic vesicles were analyzed by detergent solubilization followed by size fractionation or immunoprecipitation. The behavior of six synaptic vesicle membrane proteins as well as a plasma membrane protein was monitored by Western blotting. Solubilization of synaptic vesicle membranes in CHAPS resulted in the recovery of a large protein complex that included SV2, p65, p38, vesicle-associated membrane protein, and the vacuolar proton pump. Solubilization in octylglucoside resulted in the preservation of interactions between SV2, p38, and rab3A, while solubilization of synaptic vesicles with Triton X-100 resulted in two predominant interactions, one involving p65 and SV2, and the other involving p38 and vesicle-YNAPTIC transmission is the primary basis of intercellular communication within the nervous system. Central to this process are the synaptic vesicles respon sible for the storage and exocytotic release ofneurotransmitter. The life cycle of a synaptic vesicle includes steps of biogenesis, exocytosis, and membrane recycling (for reviews see Kelly, 1988 ; Sudhof and Jahn, 1991; Trimble et al., 1991). Synaptic vesicle components are synthesized in the cell body and transported down the axon to the presynaptic nerve terminal. Within the terminal, the vesicles are loaded with neurotransmitter and stored either adjacent to the presynaptic plasma membrane at the active zone, or in clusters associated with cytoskeletal elements. Stimulation of the nerve terminal results in calcium influx which effects fusion of the synaptic vesicles with the plasma membrane. After exocytosis, synaptic vesicle membrane components are selectively recovered from the presynaptic plasma membrane by endocytosis and are recycled within the nerve terminal into mature synaptic vesicles. A molecular description of these membrane trafficking events and their physiological regulation is required for a more detailed understanding of synaptic transmission. One approach toward understanding the molecular mechanisms involved in synaptic transmission has been to identify and characterize the components of the synaptic vesicle membrane. A number of proteins are localized to synaptic

… of Sciences of the United States …, 1993

Proceedings of the National Academy of Sciences, 2011

Sensory and signaling pathways are exquisitely organized in primary cilia. Bardet-Biedl syndrome ... more Sensory and signaling pathways are exquisitely organized in primary cilia. Bardet-Biedl syndrome (BBS) patients have compromised cilia and signaling. BBS proteins form the BBSome, which binds Rabin8, a guanine nucleotide exchange factor (GEF) activating the Rab8 GTPase, required for ciliary assembly. We now describe serumregulated upstream vesicular transport events leading to centrosomal Rab8 activation and ciliary membrane formation. Using live microscopy imaging, we show that upon serum withdrawal Rab8 is observed to assemble the ciliary membrane in ∼100 min. Rab8-dependent ciliary assembly is initiated by the relocalization of Rabin8 to Rab11-positive vesicles that are transported to the centrosome. After ciliogenesis, Rab8 ciliary transport is strongly reduced, and this reduction appears to be associated with decreased Rabin8 centrosomal accumulation. Rab11-GTP associates with the Rabin8 COOH-terminal region and is required for Rabin8 preciliary membrane trafficking to the centrosome and for ciliogenesis. Using zebrafish as a model organism, we show that Rabin8 and Rab11 are associated with the BBS pathway. Finally, using tandem affinity purification and mass spectrometry, we determined that the transport protein particle (TRAPP) II complex associates with the Rabin8 NH 2 -terminal domain and show that TRAPP II subunits colocalize with centrosomal Rabin8 and are required for Rabin8 preciliary targeting and ciliogenesis.

The Journal of cell …, 1992

Potential interactions between membrane components of rat brain synaptic vesicles were analyzed b... more Potential interactions between membrane components of rat brain synaptic vesicles were analyzed by detergent solubilization followed by size fractionation or immunoprecipitation. The behavior of six synaptic vesicle membrane proteins as well as a plasma membrane protein was monitored by Western blotting. Solubilization of synaptic vesicle membranes in CHAPS resulted in the recovery of a large protein complex that included SV2, p65, p38, vesicle-associated membrane protein, and the vacuolar proton pump. Solubilization in octylglucoside resulted in the preservation of interactions between SV2, p38, and rab3A, while solubilization of synaptic vesicles with Triton X-100 resulted in two predominant interactions, one involving p65 and SV2, and the other involving p38 and vesicle-YNAPTIC transmission is the primary basis of intercellular communication within the nervous system. Central to this process are the synaptic vesicles respon sible for the storage and exocytotic release ofneurotransmitter. The life cycle of a synaptic vesicle includes steps of biogenesis, exocytosis, and membrane recycling (for reviews see Kelly, 1988 ; Sudhof and Jahn, 1991; Trimble et al., 1991). Synaptic vesicle components are synthesized in the cell body and transported down the axon to the presynaptic nerve terminal. Within the terminal, the vesicles are loaded with neurotransmitter and stored either adjacent to the presynaptic plasma membrane at the active zone, or in clusters associated with cytoskeletal elements. Stimulation of the nerve terminal results in calcium influx which effects fusion of the synaptic vesicles with the plasma membrane. After exocytosis, synaptic vesicle membrane components are selectively recovered from the presynaptic plasma membrane by endocytosis and are recycled within the nerve terminal into mature synaptic vesicles. A molecular description of these membrane trafficking events and their physiological regulation is required for a more detailed understanding of synaptic transmission. One approach toward understanding the molecular mechanisms involved in synaptic transmission has been to identify and characterize the components of the synaptic vesicle membrane. A number of proteins are localized to synaptic

… of Sciences of the United States …, 1993

Proceedings of the National Academy of Sciences, 2011

Sensory and signaling pathways are exquisitely organized in primary cilia. Bardet-Biedl syndrome ... more Sensory and signaling pathways are exquisitely organized in primary cilia. Bardet-Biedl syndrome (BBS) patients have compromised cilia and signaling. BBS proteins form the BBSome, which binds Rabin8, a guanine nucleotide exchange factor (GEF) activating the Rab8 GTPase, required for ciliary assembly. We now describe serumregulated upstream vesicular transport events leading to centrosomal Rab8 activation and ciliary membrane formation. Using live microscopy imaging, we show that upon serum withdrawal Rab8 is observed to assemble the ciliary membrane in ∼100 min. Rab8-dependent ciliary assembly is initiated by the relocalization of Rabin8 to Rab11-positive vesicles that are transported to the centrosome. After ciliogenesis, Rab8 ciliary transport is strongly reduced, and this reduction appears to be associated with decreased Rabin8 centrosomal accumulation. Rab11-GTP associates with the Rabin8 COOH-terminal region and is required for Rabin8 preciliary membrane trafficking to the centrosome and for ciliogenesis. Using zebrafish as a model organism, we show that Rabin8 and Rab11 are associated with the BBS pathway. Finally, using tandem affinity purification and mass spectrometry, we determined that the transport protein particle (TRAPP) II complex associates with the Rabin8 NH 2 -terminal domain and show that TRAPP II subunits colocalize with centrosomal Rabin8 and are required for Rabin8 preciliary targeting and ciliogenesis.