Defects in the long-range transport of membrane organelles and macromolecules underlie the pathogenesis and progression of many diseases including neurodegenerative disorders, mental retardation and traumatic brain injury. Long-range transport takes place along the microtubule (MT) cytoskeleton and is driven by the kinesin and dynein motors. Understanding the mechanisms of MT-dependent transport is essential for the development of therapies that aim to alleviate the neurotoxicity associated with transport blockages. Despite many advances, it remains largely unknown how long-range transport is spatially and temporally controlled. This is a fundamental gap in our knowledge of cell biology and impacts understanding of many fundamental processes including how neuronal proteins navigate a geometrically complex cytoskeleton and reach their proper destinations in axons and dendrites. Spatial regulation requires modulation of the activity of MT motors and their interaction with MTs. Notably, MTs are now thought to provide guidance cues through MT-associated proteins (MAPs) and tubulin modifications, affecting the itinerary of MT motors and their cargo. Knowledge of these MT-based guidance cues, however, is rather limited and the mechanisms underlying the spatial control of MT-dependent transport are little understood. The long-term goal of this project is to understand how MT- dependent transport is regulated by septins, a family of GTP-binding proteins that associate with MTs. Our preliminary data demonstrate that septins directly modulate MT-kinesin interactions, affecting the transport of kinesins and cargo in hippocampal neurons. Here, we will investigate this unprecedented function of septins, testing the hypothesis that septins comprise a novel regulatory module for the spatial guidance of membrane traffic. Based on work in progress, we will use cell biological and in vitro cell-free approaches to mechanistically determine: 1) how septins control neuronal membrane traffic, and 2) how the motility of kinesin motors is regulated by MT-associated septins. Our studies will provide new insights into the spatial regulation of MT-dependent transport and the neuronal functions of septins, which are abnormally expressed in neurological and psychiatric disorders. In the long-term, our studies will pave the way to new therapeutic strategies that aim to restore MT-dependent transport in diseased neurons.

Public Health Relevance

Neurodegenerative diseases (e.g., Alzheimer's, Parkinson's, ALS) and brain injury due to physical trauma or stroke are characterized by blockages in the transport of proteins inside neurons. This project aims to advance our knowledge of how the long-range transport of proteins is regulated by a unique family of proteins called septins. The proposed research will generate basic knowledge, which in the long-term will be used to reverse or slow down the defects in the protein transport of sick neurons.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM097664-06
Application #
9174578
Study Section
Synapses, Cytoskeleton and Trafficking Study Section (SYN)
Program Officer
Gindhart, Joseph G
Project Start
2011-08-01
Project End
2020-07-31
Budget Start
2016-08-01
Budget End
2017-07-31
Support Year
6
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Drexel University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
002604817
City
Philadelphia
State
PA
Country
United States
Zip Code
19102
Karasmanis, Eva P; Phan, Cat-Thi; Angelis, Dimitrios et al. (2018) Polarity of Neuronal Membrane Traffic Requires Sorting of Kinesin Motor Cargo during Entry into Dendrites by a Microtubule-Associated Septin. Dev Cell 46:518-524
Spiliotis, Elias T (2018) Spatial effects?-?site-specific regulation of actin and microtubule organization by septin GTPases. J Cell Sci 131:
Dolat, Lee; Spiliotis, Elias T (2016) Septins promote macropinosome maturation and traffic to the lysosome by facilitating membrane fusion. J Cell Biol 214:517-27
Bai, Xiaobo; Karasmanis, Eva P; Spiliotis, Elias T (2016) Septin 9 interacts with kinesin KIF17 and interferes with the mechanism of NMDA receptor cargo binding and transport. Mol Biol Cell 27:897-906
Angelis, Dimitrios; Spiliotis, Elias T (2016) Septin Mutations in Human Cancers. Front Cell Dev Biol 4:122
Spiliotis, E T; Karasmanis, E P; Dolat, L (2016) Fluorescence microscopy of actin- and microtubule-associated septins in mammalian cells. Methods Cell Biol 136:243-68
Smith, Clayton; Dolat, Lee; Angelis, Dimitrios et al. (2015) Septin 9 Exhibits Polymorphic Binding to F-Actin and Inhibits Myosin and Cofilin Activity. J Mol Biol 427:3273-3284
Angelis, Dimitrios; Karasmanis, Eva Pauline; Bai, Xiaobo et al. (2014) In silico docking of forchlorfenuron (FCF) to septins suggests that FCF interferes with GTP binding. PLoS One 9:e96390
Dolat, Lee; Hu, Qicong; Spiliotis, Elias T (2014) Septin functions in organ system physiology and pathology. Biol Chem 395:123-41
Dolat, Lee; Hunyara, John L; Bowen, Jonathan R et al. (2014) Septins promote stress fiber-mediated maturation of focal adhesions and renal epithelial motility. J Cell Biol 207:225-35

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