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Determining structural mechanisms of transporters and signaling networks in cellular stress responses


EMSL Project ID
50748

Abstract

Rapidly dividing or invasive cells tied to diseases such as cancer, fungal or bacterial infection require responses that enable their survival during nutrient deprivation and drug therapy. Tightly controlled networks of transporters help regulate these stress responses, contributing to the severe mortality of several hard to treat diseases. We focus on several of these diseases including two fungal transporters (Ycf1 and Aus1), two bacterial transporters (TmrAB and BmrCD) as well as two regulators of nutrient stress responses via nitric oxide signaling (CD47 and sGC). We aim to use cryo-EM to determine structures of these transporters and signaling proteins to enable 1) a deeper understanding of their biology and 2) potential drug development to combat disease. We build from our extensive preliminary cryo-EM data where we collected datasets for Ycf1, Aus1, and BmrCD at PNCC to yield 3D structures of two (Ycf1 and BmrCD) and identified conditions for future data collection (Aus1). Together, the projects outline represent several stages of the cryo-EM pipeline ranging from the need for additional data enabling a near atomic resolution 3D structure within the next 3-6 months (Ycf1 and BmrCD), to optimization of sample preparation for cryo-EM data collection (Aus1 and BmrCD), as well as very early stage screening of cryopreservation and sample behavior (sGC and CD47). This proposal represents a long-term goal to establish cryo-EM at the University of Arizona, with a focus on expanding expertise to new trainees, and is in addition the basis for the Tomasiak laboratory’s long term interest in nutrient transporter and signaling structural biology.

Project Details

Start Date
2019-04-15
End Date
2021-03-17
Status
Closed

Team

Principal Investigator

Thomas Tomasiak
Institution
University of Arizona

Team Members

William Montfort
Institution
University of Arizona

Cheng-Yu Chen
Institution
University of Arizona

Amanda Johnson
Institution
University of Arizona

Sarah Young
Institution
University of Arizona

Nitesh Khandelwal
Institution
University of Arizona

Nancy Meyer
Institution
Oregon Health & Science University

Cinthia Millan
Institution
University of Arizona

Tarjani Thaker
Institution
University of Arizona

Irina El Khoury
Institution
Environmental Molecular Sciences Laboratory

Related Publications

José D. Faraldo-Goméz, Hassane S. Mchaourab, Smriti Mishra, Michael Mohan, Qingyu Tang, Tarjani M. Thaker, Thomas M. Tomasiak, Wenchang Zhou. 2021. "Asymmetric drug binding in an ATP-loaded inward-facing state of an ABC transporter." Nature Chemical Biology https://doi.org/10.1038/s41589-021-00936-x