Deciphering the Circadian Interactome

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.

Circadian Regulation of Growth

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.

Tool development for Plants

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.



Sorkin ML, Tzeng SC, Romanowski A, Kahle N, Bindbeutel R, Hiltbrunner A, Yanovsky MJ, Evans BS, Nusinow DA COR27/28 Regulate the Evening Transcriptional Activity of the RVE8-LNK1/2 Circadian Complex bioRxiv 492168 (2022); doi: 10.1101/2022.05.16.492168

Gao M, Geng F, Klose C, Staudt AM, Huang H, Nguyen D, Lan H, Mockler TC, Nusinow DA, Hiltbrunner A, Schäfer E, Wigge PA, and Jaeger KE Phytochromes Measure Photoperiod In Brachypodium bioRxiv 697169 (2019); doi: 10.1101/697169


Beltrán J, Steiner PJ, Bedewitz M, Wei S, Peterson FC, Li Z, Hughes BE, Hartley Z, Robertson NR, Medina-Cucurella AV, Baumer ZT, Leonard AC, Park SY, Volkman BF, Nusinow DA, Zhong W, Wheeldon I, Cutler SR, Whitehead TA. Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds Nature Biotechnology (2022) Pub. June 20, 2022 doi: 10.1038/s41587-022-01364-5.

Steiner PJ, Swift SD, Bedewitz M, Wheeldon I, Cutler SR, Nusinow DA, Whitehead TA. A Closed Form Model for Molecular Ratchet-Type Chemically Induced Dimerization Modules Biochemistry (2022) Article ASAP doi: 10.1021/acs.biochem.2c00172

4. Zhang N, Pazouki L, Nguyen H, Jacobshagen S, Bigge BM, Xia M, Mattoon EM, Klebanovych A, Sorkin M, Nusinow DA, Avasthi P, Czymmek KJ, Zhang R.Comparative Phenotyping of Two Commonly Used Chlamydomonas reinhardtii Background Strains: CC-1690 (21gr) and CC-5325 (The CLiP Mutant Library Background) Plants. (2022) 11(5):585. doi: 10.3390/plants11050585

Sorkin ML, Nusinow DA Using Tandem Affinity Purification to Identify Circadian Clock Protein Complexes from Arabidopsis Methods Mol Biology (2022) 2398:189-203. doi: 10.1007/978-1-0716-1912-4_15. PubMed PMID: 34674177


Griffiths M, Delory BM, Jawahir V, Wong KM, Bagnall GC, Dowd TG, Nusinow DA, Miller AJ, Topp CN Optimisation of root traits to provide enhanced ecosystem services in agricultural systems: A focus on cover crops Plant Cell Environment (2021) Dec 16;. doi: 10.1111/pce.14247. [Epub ahead of print] PubMed PMID: 34914117

Anderson CM, Mattoon EM, Zhang N, Becker E, McHargue W, Yang J, Patel D, Dautermann O, McAdam SAM, Tarin T, Pathak S, Avenson TJ, Berry J, Braud M, Niyogi KK, Wilson M, Nusinow DA, Vargas R, Czymmek KJ, Eveland AL, Zhang R High light and temperature reduce photosynthetic efficiency through different mechanisms in the C4 model Setaria viridis Communcation Biology (2021) Sep 16;4(1):1092. doi: 10.1038/s42003-021-02576-2. PubMed PMID: 34531541; PubMed Central PMCID: PMC8446033

Pardi SA, Nusinow DA Out of the Dark and Into the Light: A New View of Phytochrome Photobodies Front Plant Science 2021;12:732947. doi: 10.3389/fpls.2021.732947. eCollection 2021. Review. PubMed PMID: 34531891; PubMed Central PMCID: PMC8438518.

Sorkin Ml and Nusinow DA Time Will Tell: Intercellular Communication in the Plant Clock Trends in Plant Science (2021) doi: 10.1016/j.tplants.2020.12.009

Wilson ME, Tzeng SC, Augustin MM, Meyer M, Jiang X, Choi J, Bulter J, Evans B, Kutchan TM, Nusinow DA Quantitative Proteomics and Phosphoproteomics Support a Role for Mut9-Like Kinases in Multiple Metabolic and Signaling Pathways in Arabidopsis Molecular and Cellular Proteomics (2021) e100063 doi: 10.1016/j.mcpro.2021.100063


Odipio J, Getu B, Chauhan RD, Alicai T, Bart R, Nusinow DA, Taylor NJ Transgenic overexpression of endogenous FLOWERING LOCUS T-like gene MeFT1 produces early flowering in cassava PLoS One (2020) 15(1): e0227199 doi: 10.1371/journal.pone.0227199

Chen WW, Takahashi N, Hirata Y, Ronald J, Porco S, Davis SJ, Nusinow DA, Kay SA, Mas P A mobile ELF4 delivers circadian temperature information from shoots to roots Nature Plants (2020) Apr;6(4):416-426. doi: 10.1038/s41477-020-0634-2

Nimmo HG, Laird J, Bindbeutel R, Nusinow DA The evening complex is central to the difference between the circadian clocks of Arabidopsis thaliana shoots and roots Physiologia Plantarum (2020) Apr 17;. doi: 10.1111/ppl.13108/p>

Mamidi S, Healy AS, Huang P, Grimwood J, Jenkins J, Barry K, Sreedasyam A, Shu S, Lovell JT, Feldman M, Wu J, Yu Y, Chen C, Johnson J, Sakakibara H, Kiba T, Sakurai T, Tavares R, Nusinow DA, Baxter I, Schmutz J, Brutnell TP, Kellogg EAA genome resource for green millet Setaria viridis enables discovery of agronomically valuable loci Nature Biotechnology (2020) 38, pages1203–1210 doi: 10.1038/s41587-020-0681-2>


Huang H, McLoughlin KE, Sorkin ML, Burgie ES, Bindbeutel RK, Vierstra RD, Nusinow DA PCH1 regulates light, temperature, and circadian signaling as a structural component of phytochrome B-photobodies in Arabidopsis Proceedings of the National Academy of Sciences (2019) doi: 10.1073/pnas.1818217116

Kim SC, Nusinow DA, Sorkin ML, Pruneda-Paz J, Wang X Interaction and regulation between lipid mediator phosphatidic acid and circadian clock regulators in Arabidopsis. Plant Cell (2019) doi: 10.1105/tpc.18.00675


Song YH, Kubota A, Kwon MS, Covington MF, Lee N, Taagen ER, Cintrón DL, Hwang DY, Reiko Akiyama R, Hodge SK, Huang H, Nguyen NH, Nusinow DA, Millar AJ, Shimizu KK, and Imaizumi T. Molecular basis of flowering under natural long-day conditions in Arabidopsis. Nature Plants (2018) doi: 10.1038/s41477-018-0253-3

Tovar JC, Hoyer JS, Lin A, Tiekling A, Callen ST, Castillo SE, Miller M, Monica Tessman M, Fahlgren N, Carrington JC, Nusinow DA, Gehan MA Raspberry Pi Powered Imaging for Plant Phenotyping Applications in Plant Sciences (2018) doi: 10.1002/aps3.1031

Khan MA, Castro-Guerrero NA, McInturf SA, Nguyen NT, Dame AN, Wang J, Bindbeutel RK, Joshi T, Jurisson SS, Nusinow DA, and Mendoza-Cozatl DG Changes in iron availability in Arabidopsis are rapidly sensed in the leaf vasculature and impaired sensing leads to opposite transcriptional programs in leaves and roots Plant, Cell and Environment (2018) doi: 10.1111/pce.13192


Hughes ME, Abruzzi KC, Allada R, Anafi R, Arpat AB, Asher G, Baldi P, de Bekker C, Bell-Pedersen D, Blau J, Brown S, Ceriani MF, Chen Z, Chiu JC, Cox J, Crowell AM, DeBruyne JP, Dijk DJ, DiTacchio L, Doyle FJ, Duffield GE, Dunlap JC, Eckel-Mahan K, Esser KA, FitzGerald GA, Forger DB, Francey LJ, Fu YH, Gachon F, Gatfield D, de Goede P, Golden SS, Green C, Harer J, Harmer S, Haspel J, Hastings MH, Herzel H, Herzog ED, Hoffmann C, Hong C, Hughey JJ, Hurley JM, de la Iglesia HO, Johnson C, Kay SA, Koike N, Kornacker K, Kramer A, Lamia K, Leise T, Lewis SA, Li J, Li X, Liu AC, Loros JJ, Martino TA, Menet JS, Merrow M, Millar AJ, Mockler T, Naef F, Nagoshi E, Nitabach MN, Olmedo M, Nusinow DA, Ptáček LJ, Rand D, Reddy AB, Robles MS, Roenneberg T, Rosbash M, Ruben MD, Rund SSC, Sancar A, Sassone-Corsi P, Sehgal A, Sherrill-Mix S, Skene DJ, Storch KF, Takahashi JS, Ueda HR, Wang H, Weitz C, Westermark PO, Wijnen H, Xu Y, Wu G, Yoo SH, Young M, Zhang EE, Zielinski T, Hogenesch JB. Guidelines for Genome-Scale Analysis of Biological Rhythms Journal of Biological Rhythms (2017) doi: 10.1177/0748730417728663

Odipio J, Alicai T, Ingelbrecht I, Nusinow DA, Bart R, and Taylor N. Efficient CRISPR/CAS9 Genome Editing of Phytoene desaturase in Cassava Frontiers (2017): p1780. doi: 10.3389/fpls.2017.01780

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 Plant Direct (2017) doi:10.1002/pld3.18

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

Dmitri A. Nusinow, Ph.D.

Principal Investigator
meter (at)
ORCID ID: 0000-0002-0497-1723

Dong-Yeon Lee, Ph.D.

Staff Scientist
Dlee (at)
ORCID ID: Coming Soon

Vanessica Jawahir, Ph.D.

Postdoctoral Fellow
VJawahir (at)
ORCID ID: Coming Soon

Rebecca Bindbeutel, M.S.

Administrative Lab Manager
RBindbeutel (at)
ORCID ID: 0000-0001-7994-2073

Yuan Wang, Ph.D.

Project Manager and Intergrator, APT Program
ywang (at)
ORCID ID: Coming Soon

Maria Sorkin, Ph.D.

NSF GRFP Fellow, Washington University Graduate
MSorkin (at)
ORCID ID: 0000-0002-3897-2636

Sarah Pardi

Washington University Graduate Student
SPardi (at)
ORCID ID: 0000-0002-6881-7012

Kristen Edgeworth

Washington University Graduate Student
KEdgeworth (at)
ORCID ID: Coming Soon

Ashton Kish

Laboratory technician
akish (at)
ORCID ID: Coming Soon

Adele Ayers

Laboratory Assistant
AAyers (at)
ORCID ID: Coming Soon
Lab Alumni


Nathan Swyers, Ph.D.,M6P Therapeutics, Missouri

Austin Morgan,(Indiana Biosciences Research Institute, Indiana)

Eileen Kosola,(University of Minnesota, Twin Cities, MN)


Sunita Pathak M.S., (Graduate Student, Duke University, North Carolina)


Rachel Tavares, (Graduate Student, U. Mass Amherst, Massachusetts)

Katherine Benza, (Graduate Student, Washington University in St. Louis, Missourt)


Dr. He Huang, (Staff Scientist, Vanderbilt University, Tennessee)

Shara Chopra, Penn State University, NSF-REU

Abigail Hunt, Holt High School, Independent Intern


Sarah Huss, (Masters Program, University of Illinois, Urbana-Champaign)

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'Laan Pittman, Washington University, St. Louis, uSTAR Program (Masters program, Chemistry, George Washington University)


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