1991 — 1992 |
Wente, Susan R. |
F32Activity Code Description: To provide postdoctoral research training to individuals to broaden their scientific background and extend their potential for research in specified health-related areas. |
In Vitro Assembly of Nuclear Pore Complex |
0.943 |
1994 — 2019 |
Wente, Susan R. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. R37Activity Code Description: To provide long-term grant support to investigators whose research competence and productivity are distinctly superior and who are highly likely to continue to perform in an outstanding manner. Investigators may not apply for a MERIT award. Program staff and/or members of the cognizant National Advisory Council/Board will identify candidates for the MERIT award during the course of review of competing research grant applications prepared and submitted in accordance with regular PHS requirements. |
Molecular Analysis of Nuclear Pore Complex Function
[unreadable] DESCRIPTION (provided by applicant): Nuclear pore complexes (NPCs) form the site for entry and exit from the nucleus. The most outstanding issue in the nuclear transport field involves delineating how cargo crosses the aqueous NPC channel. Translocation thought to be is based on physical interactions between soluble shuttling transport factors and the NPC. How these interactions result in movement across the NPC is unknown. The long-range goal of this project is to understand the order of events at the NPC during nuclear transport.Our specific aims will analyze the mechanism and regulation of transport factor-NPC interaction during protein and mRNA transport. The first aim builds on our recent discovery that mRNA export requires a nuclear enzymatic pathway that converts soluble inositol 1,4,5-trisphosphate to inositol hexakisphosphate (IP6). We hypothesize that IP6 production influences events at the NPC. IP6 binding, two-hybrid and genetic strategies will be used to identify IP6 targets in S. cerevisiae and pinpoint the step in the mRNA export pathway that requires IP6 production. In the second aim, we will test specific hypotheses for the role of the essential mRNA export factor Gle1 in yeast and human cells. IP6 production is required for Gle1 function. We will investigate whether Gle1 shuttles between the nucleus and cytoplasm, and determine the network of protein-protein interactions that mediate Gle1 shuttling, NPC localization, and mRNA translocation. Finally, we will initiate new studies to test models for the NPC translocation mechanism. Yeast mutant strains will be identified that harbor a minimal repertoire of NPC transport factor binding sites, or that harbor a transport factor defective for NPC interaction. By determining the rates of nuclear import /export and the transport arrest points at the NPC, we aim to reveal how binding at the NPC results in translocation.Knowledge of how the nuclear accessibility of molecules can be selectively targeted or inhibited will be essential for designing therapeutic strategies, and understanding viral proliferation/pathogenesis. Transport factors and nucleoporins are both targets for viral inhibition of cellular function and mediators of viral RNA export. Defects in inositol signaling pathways are also associated with disease states including cancer cell growth, inflammation, and neurotransmission. Thus, these studies have direct health relatedness.
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1998 — 2011 |
Wente, Susan R. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Regulation of Nuclear Pore Complex Assembly
DESCRIPTION (provided by applicant): Nuclear pore complexes (NPCs) are large proteinaceous assemblies that provide the only known portals for exchanging macromolecules between the nucleus and cytoplasm. This includes the movement of small molecules and the selective, facilitated transport of large proteins and RNAs. Faithful, continuous NPC assembly is key for maintaining normal physiological function and is closely tied to proper cell division. Understanding how NPC biogenesis can be selectively inhibited may be key for designing strategies to inhibit cell growth, for example in oncogenesis. However, the molecular pathway of NPC assembly remains largely undefined. We hypothesize that NPC biogenesis is based on a step-wise mechanism, including a novel vesicle-mediated targeting of NPC-associated proteins to the outer nuclear membrane. The factors that mediate each assembly step are unknown. The long-range goal of this project is to elucidate the molecular sequence of events required for NPC formation. Our specific aims will address the assembly mechanism using the yeast S. cerevisiae as a model. In the first aim, we will identify essential factors required for NPC biogenesis. We have isolated a large collection of temperature sensitive NPC assembly mutants with a fluorescence-based screen. A battery of microscopy and genetic assays will pinpoint mutants with direct defects in new assembly. The second aim builds on our recent discovery that RanGTPase cycle mutants block new NPC assembly by disrupting the targeting of Nup-containing vesicles to the nuclear membrane. We hypothesize that RanGTP is required for both Nup import and for the novel vesicle-mediated step. To investigate this, the accumulated vesicles will be purified and associated proteins tested for assembly roles. Genetic and cell biology strategies will also be used to analyze interactions with Ran. Finally, we will initiate new studies to develop yeast in vitro NPC assembly assay. This is based on our hypothesis that the Nup-containing vesicles are a functional assembly intermediate. Direct tests for the roles of specific proteins in NPC assembly will be conducted and allow for the first time the coupling of genetic, biochemical, and structural analysis of NPC biogenesis in a single system.
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2010 |
Wente, Susan R. |
G20Activity Code Description: To provide funds for major repair, renovation, and modernization of existing research facilities. These facilities may be the clinical research facilities, animal research facilities, and other related research facilities. |
Vantage:Consolidation to Create the Vanderbilt Technologies For Advanced Genomics
DESCRIPTION (provided by applicant): Recognizing the importance of translating advances in genome science to improved human health, Vanderbilt University School of Medicine has made a commitment to provide state-of-the art laboratory environments and technology through shared resource facilities. To that end, the purpose of this grant application is to optimize an environment conducive to collaboration and acceleration of new biological and clinical discoveries through consolidation of multiple currently physically separate research resources into a centrally located shared resource: VANTAGE (VANderbilt Technologies for Advanced GEnomics). Through complete modernization of an existing 9,316 Net Square Feet of aging, poorly-utilized laboratory space, this proposal will create a physical home for the Genome Technology Core (GTC, a collaborative core for development of next-generation technologies) and BioVU (the Vanderbilt DNA Repository currently housed within the DNA Resources Core), and will enable the consolidation and expansion of four existing core facilities: DNA Resources Core, DNA Sequencing Core, Vanderbilt Microarray Shared Resource (VMSR) and the Flow Cytometry Core. Once renovation is complete, each of these distinct research resources will share collaborative space, thereby creating an institutional nexus of discovery in genomics research by extending and strengthening already successful collaborations. VANTAGE will enable technical synergy, eliminate unnecessary duplication and accelerate discovery in clinical and basic science across the institution. In particular, this renovation will impact over 968 active grants from major Vanderbilt users involving approximately 235 million in annual direct costs.
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