Determining how plants know what time it is without a watch and which season they are in without a calendar.
Plants use an endogenous time keeping mechanism known as the circadian clock to regulate the timing of metabolism, physiology and development. However, the basic composition of the circadian system is unknown and how the clock is connected to other pathways in unclear. We combine affinity purification, mass spectrometry, and genetics to identify, dissect, and define the protein complexes within the clock.
The circadian clock regulates myriad processes in plants. We are particularly interested in how plants adapt their growth to seasonal variation in temperature and day length. We have identified new molecules that act as conduits between the circadian system and growth pathways. We are determining the function of these key proteins with the aim of improving crop production in response to a changing environment.
In order to determine the molecular mechanisms that underlie key plant-specific processes, we are developing new tools and methods to explore protein function. This includes developing methods for rapid, facile purification of protein complexes and new methods for maintaining epitope-tagged proteins at endogenous levels and in their native context. We also develop inexpensive non-invasive imaging systems to monitor plant movement, bioluminescence and fluorescence during long-term time lapse experiments. We make the protocols, instructions, and scripts available and open-source so that we can leverage the community to improve and extend these when possible.
Huang H, Gehan M, Huss SE, Alvarez S, Lizarraga C, Gruebbling EL, Gierer J, Naldrett MJ, Bindbeutel R, Evans BS, Mockler TC, Nusinow DA Cross-Species Complementation Reveals Conserved Functions For EARLY FLOWERING 3 Between Monocots And Dicots bioRxiv 131185; doi:10.1101/131185
Lee CM, Adamchek C, Feke A, Nusinow DA, and Gendron JM Mapping Protein–Protein Interactions Using Affinity Purification and Mass Spectrometry Methods In Molecular Biology 1610, no. M110 (2017): doi:10.1007/978-1-4939-7003-2_15
Huang H and Nusinow DA Tandem Purification of His6-3x FLAG Tagged Proteins for Mass Spectrometry from Arabidopsis. Bio-protocol (2016): doi:10.21769/BioProtoc.2060
Huang H and Nusinow DA Into the Evening: Complex Interactions in the Arabidopsis circadian clock. Trends in Genetics (2016): doi:10.1016/j.tig.2016.08.002
Mutka AM, Fentress SJ, Sher JW, Berry JC, Pretz C, Nusinow DA and Bart R Quantitative, image-based phenotyping methods provide insight into spatial and temporal dimensions of plant disease. Plant Physiology (2016): doi:10.1104/pp.16.00984
Huang H, Alvarez S, and Nusinow DA Data on the Identification of Protein Interactors with the Evening Complex and PCH1 in Arabidopsis Using Tandem Affinity Purification and Mass Spectrometry (TAP–MS) Data in Brief (2016): doi:10.1016/j.dib.2016.05.014
Huang H, Yoo CY, Bindbeutel RK, Goldsworthy J, Tielking A, Alvarez S, Naldrett M, Evans B, Chen M, and Nusinow DA. PCH1 integrates circadian and light-signaling pathways to control photoperiod-responsive growth in Arabidopsis. eLife. (2016) doi:10.7554/eLife.13292.
Huang H, Alvarez S, Bindbeutel RK, Shen Z, Naldrett MJ, Evans BS, Briggs SP, Hicks LM, Kay SA, and Nusinow DA. Identification of evening complex associated proteins in Arabidopsis by affinity purification and mass spectrometry. Molecular & Cellular Proteomics. (2016). doi:10.1074/mcp.M115.054064
Kaiserli E, Paldi K, O’Donnell L, Batalov O, Pedmale UV, Nusinow DA, Kay SA, Chory J. Integration of light and photoperiodic signaling in transcriptional nuclear foci. Developmental Cell (2015) doi:10.1016/j,devcel.2015.10.008
Waadt R, Manalansan B, Rauniyar N, Munemasa S, Booker MA, Brandt B, Waadt C, Nusinow, DA, Kay SA, Kunz HH, Schumacher K, DeLong A, Yates JR, and Schroeder JI. Identification of OST1-Interacting Proteins Reveals Interactions with SnRK2-Type Protein Kinases and with PP2A-Type Protein Phosphatases That Function in ABA Responses. Plant Physiology (2015) doi:10.1104/pp.15.00575
Chow BY, Helfer A, Nusinow DA, Kay SA. ELF3 recruitment to the PRR9 promoter requires other Evening Complex members in the Arabidopsis circadian clock. Plant Signaling and Behavior (2012) doi:10.4161/psb.18766
Nusinow DA, Helfer A, Hamilton EE, King JJ, Imaizumi T, Schultz TF, Farre EM, Kay SA. The ELF4-ELF3-LUX Complex Links the Circadian Clock to Diurnal Control of Hypocotyl Growth. Nature (2011) doi:10.1038/nature10182
Helfer A, Nusinow DA, Chow BY, Gehrke AR, Bulyk ML, Kay SA. LUX ARRHYTHMO Encodes a Nighttime Repressor of Circadian Gene Expression in the Arabidopsis Core Clock. Current Biology doi:10.1016/j.cub.2010.12.021
Mayla Ayers, Harris Stowe University, NSF–REU
Abigail Hunt, Holt High School, Independent Intern
David Stroshein, Lawrence Technological University, NSF-REU (Graduate School, U of Arizona)
J'Loan Pittman, Washington University, St. Louis, uSTAR Program
Dhruv Patel, Cornell, NSF-REU (Graduate School, UC Berkeley)
Hannah Lucas, U. of Wisconsin, Madison, NSF-REU (Graduate School, Washington U, St. Louis)
Ali Peet, St. Joseph’s Academy, Independent Intern
WIll Wolfe, John Burroughs High School, Independent Intern
Jessica Goldsworthy, Michigan State University, NSF-REU (Pharm D. Program, Ferris State University)
Tom Liu, Ladue High School, STARS Intern
Allison Tielking, MICDS, Independent Intern
Morgan Clark, Western Illinois University, NSF-REU (Chemist, Covance)
Elle Gruebbeling, Timberland High School, STARS Intern
Savannah Est, Holt High School, Independent Intern