Land plants evolved from green algae predecessors around 470 million years ago, a key evolutionary step facilitated by their novel ability to perceive positional information and translate this into 3D growth patterns. The exact mechanism(s) for positional signaling in plants is still unknown. DEFECTIVE KERNEL 1 (DEK1), a large multi-domain protein with membrane-anchored calpain, has been proposed as a key regulator of oriented cell division and positional cell fate specification in land plants.

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We aim to contribute to the understanding of DEK1 function by determining the structure of DEK1 protein and to identify its downstream targets,including in vivo substrates of the calpain protease. We have proposed that DEK1, a 240 kDa protein have evolved from ancient TML-calpains (existed 1.5 billion yrs ago) by fusion of genes encoding a Major facilitator superfamily membrane protein, an LG3 domain containing protein and a calpain. We aim to express and purify the full-length DEK1 recombinant protein as well as its calpain catalytic core, to determine the structure of DEK1 using cryo-EM and X-ray crystallography. Experimental systems for functional dissection of DEK1 using genetics and cell biology include the moss Physcomitrella patens and Arabidopsis thaliana.

Project team members

VIKTOR DEMKO Norwegian University of Life Sciences

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WENCHE JOHANSEN Hedmark University of Applied Sciences

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AKO EUGENE AKO Hedmark University of Applied Sciences

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PIERRE-FRANCOIS PERROUD Philipps University of Marburg

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Collaborators

Selected publications

THE DEK1 CALPAIN LINKER FUNCTIONS IN THREE-DIMENSIONAL BODY PATTERING IN PHYSCOMITRELLA PATENS. PLANT PHYSIOL

Johansen W, Eugene A, Demko V, Perroud PF, Rensing S, Khaleel Mekhlif A and Olsen OA(2016), in press
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THE PHENOTYPE OF THE CRINKLY4 DELETION MUTANT OF PHYSCOMITRELLA PATENS SUGGESTS A BROAD ROLE IN DEVELOPMENTAL REGULATION IN EARLY LAND PLANTS.

Demko V, Ako E, Perroud PF, Quatrano R, Olsen OA. Planta. 2016 Jul;244(1):275-84. doi: 10.1007/s00425-016-2526-2. Epub 2016 Apr 21. PMID: 27100110

DEK1; MISSING PIECE IN PUZZLE OF PLANT DEVELOPMENT.

Olsen OA, Perroud PF, Johansen W, Demko V. Trends Plant Sci. 2015 Feb;20(2):70-1. doi: 10.1016/j.tplants.2015.01.003. Epub 2015 Jan 20. PMID: 25612461

GENETIC ANALYSIS OF DEFECTIVE KERNEL1 LOOP FUNCTION IN THREE-DIMENSIONAL BODY PATTERNING IN PHYSCOMITRELLA PATENS.

Demko V, Perroud PF, Johansen W, Delwiche CF, Cooper ED, Remme P, Ako AE, Kugler KG, Mayer KF, Quatrano R, Olsen OA. Plant Physiol. 2014 Oct;166(2):903-19. doi: 10.1104/pp.114.243758. Epub 2014 Sep 2. PMID: 25185121
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DEFECTIVE KERNEL 1 (DEK1) IS REQUIRED FOR THREE-DIMENSIONAL GROWTH IN PHYSCOMITRELLA PATENS.

Perroud PF, Demko V, Johansen W, Wilson RC, Olsen OA, Quatrano RS. New Phytol. 2014 Aug;203(3):794-804. doi: 10.1111/nph.12844. Epub 2014 May 21. PMID: 24844771
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THE CATALYTIC DOMAIN CYSPC OF THE DEK1 CALPAIN IS FUNCTIONALLY CONSERVED IN LAND PLANTS.

Liang Z, Demko V, Wilson RC, Johnson KA, Ahmad R, Perroud PF, Quatrano R, Zhao S, Shalchian-Tabrizi K, Otegui MS, Olsen OA, Johansen W. Plant J. 2013 Sep;75(5):742-54. doi: 10.1111/tpj.12235. Epub 2013 Jun 13. PMID: 23663131
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MASSIVE EXPANSION OF THE CALPAIN GENE FAMILY IN UNICELLULAR EUKARYOTES.

Zhao S, Liang Z, Demko V, Wilson R, Johansen W, Olsen OA, Shalchian-Tabrizi K. BMC Evol Biol. 2012 Sep 29;12:193. doi: 10.1186/1471-2148-12-193. PMID: 23020305

SUBCELLULAR LOCALIZATION AND FUNCTIONAL DOMAIN STUDIES OF DEFECTIVE KERNEL1 IN MAIZE AND ARABIDOPSIS SUGGEST A MODEL FOR ALEURONE CELL FATE SPECIFICATION INVOLVING CRINKLY4 AND SUPERNUMERARY ALEURONE LAYER1.

Tian Q, Olsen L, Sun B, Lid SE, Brown RC, Lemmon BE, Fosnes K, Gruis DF, Opsahl-Sorteberg HG, Otegui MS, Olsen OA. Plant Cell. 2007 Oct;19(10):3127-45. Epub 2007 Oct 12. PMID: 17933905
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SURFACE POSITION, NOT SIGNALING FROM SURROUNDING MATERNAL TISSUES, SPECIFIES ALEURONE EPIDERMAL CELL FATE IN MAIZE.

Gruis DF, Guo H, Selinger D, Tian Q, Olsen OA. Plant Physiol. 2006 Jul;141(3):898-909. Epub 2006 May 12. Erratum in: Plant Physiol. 2006 Dec;142(4):1771. PMID: 16698897
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MUTATION IN THE ARABIDOPISIS THALIANA DEK1 CALPAIN GENE PERTURBS ENDOSPERM AND EMBRYO DEVELOPMENT WHILE OVER-EXPRESSION AFFECTS ORGAN DEVELOPMENT GLOBALLY.

Lid SE, Olsen L, Nestestog R, Aukerman M, Brown RC, Lemmon B, Mucha M, Opsahl-Sorteberg HG, Olsen OA. Planta. 2005 Jun;221(3):339-51. Epub 2005 Jan 13.

THE MAIZE DISORGANIZED ALEURONE LAYER 1 AND 2 ( DIL1, DIL2) MUTANTS LACK CONTROL OF THE MITOTIC DIVISION PLANE IN THE ALEURONE LAYER OF DEVELOPING ENDOSPERM.

Lid SE, Al RH, Krekling T, Meeley RB, Ranch J, Opsahl-Ferstad HG, Olsen OA. Planta. 2004 Jan;218(3):370-8. Epub 2003 Oct 14. PMID: 14557881

SAL1 DETERMINES THE NUMBER OF ALEURONE CELL LAYERS IN MAIZE ENDOSPERM AND ENCODES A CLASS E VACUOLAR SORTING PROTEIN.

Shen B, Li C, Min Z, Meeley RB, Tarczynski MC, Olsen OA. Proc Natl Acad Sci U S A. 2003 May 27;100(11):6552-7. Epub 2003 May 15. PMID: 12750475
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THE CALPAIN DOMAIN OF THE MAIZE DEK1 PROTEIN CONTAINS THE CONSERVED CATALYTIC TRIAD AND FUNCTIONS AS A CYSTEINE PROTEINASE.

Wang C, Barry JK, Min Z, Tordsen G, Rao AG, Olsen OA. J Biol Chem. 2003 Sep 5;278(36):34467-74. Epub 2003 Jun 24. PMID: 12824178
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THE DEFECTIVE KERNEL 1 (DEK1) GENE REQUIRED FOR ALEURONE CELL DEVELOPMENT IN THE ENDOSPERM OF MAIZE GRAINS ENCODES A MEMBRANE PROTEIN OF THE CALPAIN GENE SUPERFAMILY.

Lid SE, Gruis D, Jung R, Lorentzen JA, Ananiev E, Chamberlin M, Niu X, Meeley R, Nichols S, Olsen OA. Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5460-5. Epub 2002 Apr 2. PMID: 11929961
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