Transition metals are critical to all forms of life, and metal ions play vital roles in a diverse array of fundamental biological processes. Metal dyshomeostasis is a central feature of a broad spectrum of human diseases, yet elucidating whether altered metal status is a cause or consequence of disease is exceedingly difficult without a basic understanding of how metals influence normal cellular functions. While there are some well-known examples of how metals can drive cellular change, there are major gaps in our understanding of how changes in metal status influence cell physiology. This gap is particularly notable for zinc. There is growing evidence that zinc changes in response to cell state, and may function in signal transduction and biological regulation. For example, massive accumulation of zinc is required for meiotic maturation of oocytes and fertilization leads to zinc sparks;stimulation of bone marrow derived mast cells leads to zinc waves. Furthermore, my lab has shown that calcium signaling events in neurons and epithelial cells lead to mobilization of zinc, raising the intriguing possibility that zinc functions as a novel second messenger. But in al of these cases, the downstream effectors, i.e. the proteins that sense changes in zinc in order to regulate cellular processes remain a complete mystery. The core hypothesis of this project is that the regulation of cellular zinc dictates occupancy of the zinc proteome, providing a novel link between dynamic metal regulation and a wide swath of cellular signaling responses. The implication of this hypothesis is that changes in zinc status - either dynamically during physiological signaling, or permanently as a consequence of disease - fine-tune the activity of hundreds, if not thousands of zinc-dependent proteins, establishing zinc as a master regulator of cellular function. Current dogma in zinc biology asserts that the ~ 2000 proteins that comprise the zinc proteome bind zinc cons

Public Health Relevance

Transition metals are critical to all forms of life and metal dyshomeostasis is a central feature of a broad spectrum of human diseases. The proposed research will elucidate the fundamental mechanisms by which zinc, a key transition metal in humans, regulates cell signaling and controls cell physiology. This will provide key insights that allow us to define how and why zinc dysregulation is associated with debilitating diseases such as diabetes, various cancers, bacterial infection, and some neurodegenerative diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
NIH Director’s Pioneer Award (NDPA) (DP1)
Project #
1DP1GM114863-01
Application #
8755503
Study Section
Special Emphasis Panel (ZRG1-BCMB-N (50))
Program Officer
Anderson, Vernon
Project Start
2014-09-30
Project End
2019-07-31
Budget Start
2014-09-30
Budget End
2015-07-31
Support Year
1
Fiscal Year
2014
Total Cost
$765,250
Indirect Cost
$265,250
Name
University of Colorado at Boulder
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
007431505
City
Boulder
State
CO
Country
United States
Zip Code
80303
Manna, Premashis; Hung, Sheng-Ting; Mukherjee, Srijit et al. (2018) Directed evolution of excited state lifetime and brightness in FusionRed using a microfluidic sorter. Integr Biol (Camb) 10:516-526
Braselmann, Esther; Wierzba, Aleksandra J; Polaski, Jacob T et al. (2018) A multicolor riboswitch-based platform for imaging of RNA in live mammalian cells. Nat Chem Biol 14:964-971
Choi, Sangyong; Hu, Ya-Mei; Corkins, Mark E et al. (2018) Zinc transporters belonging to the Cation Diffusion Facilitator (CDF) family have complementary roles in transporting zinc out of the cytosol. PLoS Genet 14:e1007262
Rodriguez, Erik A; Campbell, Robert E; Lin, John Y et al. (2017) The Growing and Glowing Toolbox of Fluorescent and Photoactive Proteins. Trends Biochem Sci 42:111-129
Specht, Elizabeth A; Braselmann, Esther; Palmer, Amy E (2017) A Critical and Comparative Review of Fluorescent Tools for Live-Cell Imaging. Annu Rev Physiol 79:93-117
Carter, Kyle P; Carpenter, Margaret C; Fiedler, Brett et al. (2017) Critical Comparison of FRET-Sensor Functionality in the Cytosol and Endoplasmic Reticulum and Implications for Quantification of Ions. Anal Chem 89:9601-9608
Carpenter, Margaret C; Palmer, Amy E (2017) Native and engineered sensors for Ca2+ and Zn2+: lessons from calmodulin and MTF1. Essays Biochem 61:237-243
Fiedler, Brett L; Van Buskirk, Steven; Carter, Kyle P et al. (2017) Droplet Microfluidic Flow Cytometer For Sorting On Transient Cellular Responses Of Genetically-Encoded Sensors. Anal Chem 89:711-719
Carpenter, Margaret C; Lo, Maria N; Palmer, Amy E (2016) Techniques for measuring cellular zinc. Arch Biochem Biophys 611:20-29