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The serine-rich N-terminal region of Arabidopsis phytochrome A is required for protein stability

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Abstract

Deletion or substitution of the serine-rich N-terminal stretch of grass phytochrome A (phyA) has repeatedly been shown to yield a hyperactive photoreceptor when expressed under the control of a constitutive promoter in transgenic tobacco or Arabidopsis seedlings retaining their native phyA. These observations have lead to the proposal that the serine-rich region is involved in negative regulation of phyA signaling. To re-evaluate this conclusion in a more physiological context we produced transgenic Arabidopsis seedlings of the phyA-null background expressing Arabidopsis PHYA deleted in the sequence corresponding to amino acids 6–12, under the control of the native PHYA promoter. Compared to the transgenic seedlings expressing wild-type phyA, the seedlings bearing the mutated phyA showed normal responses to pulses of far-red (FR) light and impaired responses to continuous FR light. In yeast two-hybrid experiments, deleted phyA interacted normally with FHY1 and FHL, which are required for phyA accumulation in the nucleus. Immunoblot analysis showed reduced stability of deleted phyA under continuous red or FR light. The reduced physiological activity can therefore be accounted for by the enhanced destruction of the mutated phyA. These findings do not support the involvement of the serine-rich region in negative regulation but they are consistent with a recent report suggesting that phyA turnover is regulated by phosphorylation.

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Abbreviations

phyA:

Phytochrome A

FR:

Far-red light

SD:

Short days

VLFR:

Very-low-fluence response

HIR:

High-irradiance response

Pr:

Red-light-absorbing form of phytochrome

Pfr:

FR-absorbing form of phytochrome

WT:

Wild type

FL:

Full-length phyA

References

  • Barnes SA, Nishizawa NK, Quaggio RB, Whitelam GC, Chua N-H (1996) Far-red light blocks greening of Arabidopsis seedlings via a phytochrome A-mediated change in plastid development. Plant Cell 8:601–615

    Article  PubMed  CAS  Google Scholar 

  • Botto JF, Sánchez RA, Whitelam GC, Casal JJ (1996) Phytochrome A mediates the promotion of seed germination by very low fluences of light and canopy shade light in Arabidopsis. Plant Physiol 110:439–444

    PubMed  CAS  Google Scholar 

  • Boylan M, Douglas N, Quail PH (1994) Dominant negative suppression of Arabidopsis photoresponses by mutant phytochrome A sequences identifies spatially discrete regulatory domains in the photoreceptor. Plant Cell 6:449–460

    Article  PubMed  CAS  Google Scholar 

  • Casal JJ, Yanovsky MJ, Luppi JP (2000) Two photobiological pathways of phytochrome A activity, only one of which shows dominant negative suppression by phytochrome B. Photochem Photobiol 71:481–486

    Article  PubMed  CAS  Google Scholar 

  • Casal JJ, Davis SJ, Kirchenbauer DJ, Viczian A, Yanovsky MJ, Clough RC, Kircher S, Jordan-Beebe ET, Schäfer E, Nagy F, Vierstra RD (2002) The serine-rich N-terminal domain of oat phytochrome A helps regulate light responses and subnuclear localization of the photoreceptor. Plant Physiol 129:1127–1137

    Article  PubMed  CAS  Google Scholar 

  • Cerdán PD, Staneloni RJ, Ortega J, Bunge MM, Rodriguez-Batiller J, Sánchez RA, Casal JJ (2000) Sustained but not transient phytochrome A signaling targets a region of a Lhcb1*2 promoter not necessary for phytochrome B action. Plant Cell 12:1203–1211

    Article  PubMed  Google Scholar 

  • Chen M, Chory J, Fankhauser C (2004) Light signal transduction in higher plants. Annu Rev Genet 38:87–117

    Article  PubMed  CAS  Google Scholar 

  • Cherry J, Hondred D, Walker J, Viestra R (1992) Phytochrome requires the 6-kDa N-terminal domain for full biological activity. Proc Natl Acad Sci USA 89:5039–5043

    Article  PubMed  CAS  Google Scholar 

  • Clough RC, Vierstra RD (1997) Phytochrome degradation. Plant Cell Environ 20:713–721

    Article  CAS  Google Scholar 

  • Emmler K, Stockhaus J, Chua N-H, Schäfer E (1995) An amino-terminal deletion of rice phytochrome A results in a dominant negative suppression of tobacco phytochrome A activity in transgenic tobacco seedlings. Planta 197:103–110

    Article  PubMed  CAS  Google Scholar 

  • Fankhauser C, Casal JJ (2004) Phenotypic characterization of a photomorphogenic mutant. Plant J 39:747–760

    Article  PubMed  CAS  Google Scholar 

  • Furuya M, Song P-S (1994) Assembly and properties of holophytochrome. In: Kendrick RE, Kronenberg GHM (eds) Photomorphogenesis in plants, 2nd edn. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 105–140

    Google Scholar 

  • Hennig L, Büche C, Eichenberg K, Schäfer E (1999) Dynamic properties of endogenous phytochrome A in Arabidopsis seedlings. Plant Physiol 121:571–578

    Article  PubMed  CAS  Google Scholar 

  • Hennig L, Büche C, Schäfer E (2000) Degradation of phytochrome A and the high irradiance response of Arabidopsis: a kinetic analysis. Plant Cell Environ 23:727–734

    Article  CAS  Google Scholar 

  • Hiltbrunner A, Viczian A, Bury E, Tscheuschler A, Kircher S, Tóth R, Honsberger A, Nagy F, Fankhauser C, Schäfer E (2005) Nuclear accumulation of the phytochrome A photoreceptor requires FHY1. Curr Biol 15:2125–2130

    Article  PubMed  CAS  Google Scholar 

  • Hiltbrunner A, Tscheuschler A, Viczian A, Kunkel T, Kircher S, Schäfer E (2006) FHY1 and FHL act together to mediate nuclear accumulation of the phytochrome A photoreceptor. Plant Cell Physiol 47:1023–1034

    Article  PubMed  CAS  Google Scholar 

  • Hisada A, Hanzawa H, Weller JL, Nagatani A, Reid JB, Furuya M (2000) Light-induced nuclear translocation of endogenous pea phytochrome A visualized by immunocytochemical procedures. Plant Cell 12:1063–1078

    Article  PubMed  CAS  Google Scholar 

  • Johnson E, Bradley M, Harberd P, Whitelam GC (1994) Photoresponses of light-grown phyA mutants of Arabidopsis. Phytochrome A is required for the perception of daylength extensions. Plant Physiol 105:141–149

    Article  PubMed  CAS  Google Scholar 

  • Jordan ET, Cherry JR, Walker JM, Vierstra RD (1995) The amino-terminus of phytochrome A contains two distinct functional domains. Plant J 9:243–257

    Article  Google Scholar 

  • Jordan ET, Marita JM, Clough RC, Vierstra RD (1997) Characterization of regions within the N-terminal 6-kilodalton domain of phytochrome A that modulate its biological activity. Plant Physiol 115:693–704

    Article  PubMed  CAS  Google Scholar 

  • Kim J-I, Shen Y, Han Y-J, Park J-E, Kirchenbauer D, Soh M-S, Nagy F, Schäfer E, Song P-S (2004) Phytochrome phosphorylation modulates light signaling by influencing the protein–protein interaction. Plant Cell 16:2629–2640

    Article  PubMed  CAS  Google Scholar 

  • Kircher S, Kozma-Bognar L, Kim L, Adam E, Harter K, Schäfer E, Nagy F (1999) Light-quality dependent nuclear import of the plant photoreceptors phytochrome A and B. Plant Cell 11:1445–1456

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Lapko VN, Jiang X-Y, Smith DL, Song P-S (1997) Posttranslational modification of oat phytochrome A: phosphorylation of a specific serine in a multiple serine cluster. Biochemistry 36:10595–10599

    Article  PubMed  CAS  Google Scholar 

  • Lapko VN, Jiang XY, Smith DL, Song PS (1999) Mass spectrometric characterization of oat phytochrome A: isoforms and posttranslational modifications. Protein Sci 8:1032–1044. http://www.biomednet.com/db/medline/99268336

    Google Scholar 

  • Mancinelli AL, Rossi F, Moroni A (1991) Cryptochrome, phytochrome and anthocyanin production. Plant Physiol 96:1079–1085

    PubMed  CAS  Google Scholar 

  • Moran R (1982) Formulae for determination of chlorophyllous pigments extracted with N,N-dimethylformamide. Plant Physiol 69:1376–1381

    Article  PubMed  CAS  Google Scholar 

  • Quail PH, Boylan MT, Parks BM, Short TW, Xu Y, Wagner D (1995) Phytochromes: photosensory perception and signal transduction. Science 268:675–680

    Article  PubMed  CAS  Google Scholar 

  • Ryu JS, Kim JI, Kunkel T, Kim BC, Cho DS, Hong SH, Kim SH, Fernandez AP, Kim Y, Alonso JM, Ecker JR, Nagy F, Lim PO, Song PS, Schäfer E, Nam HG (2005) Phytochrome-specific type 5 phosphatase controls light signal flux by enhancing phytochrome stability and affinity for a signal transducer. Cell 120:395–406

    Article  PubMed  CAS  Google Scholar 

  • Schäfer E, Nagy F (2006) Photomorphogenesis in plants and bacteria: function and signal transduction mechanisms. Springer, Berlin Heidelberg, New York

    Book  Google Scholar 

  • Schmidt TGM, Koepke J, Frank R, Skerra A (1996) Molecular interactions between the Strep-tag affinity peptide and its cognate target, streptavidin. J Mol Biol 255:753–766

    Article  PubMed  CAS  Google Scholar 

  • Schumacher K, Vafeados D, McCarthy M, Sze H, Wilkins T, Chory J (1999) The Arabidopsis det3 mutant reveals a central role for the vacuolar H+-ATPase in plant growth and development. Genes Dev 13:3259–3270

    Article  PubMed  CAS  Google Scholar 

  • Seo HS, Watanabe E, Tokutomi S, Nagatani A, Chua N-H (2004) Photoreceptor ubiquitination by COP1 E3 ligase desensitizes phytochrome A signaling. Genes Dev 18:617–622

    Article  PubMed  CAS  Google Scholar 

  • Shimizu-Sato S, Huq E, Tepperman JM, Quail PH (2002) A light-switchable gene promoter system. Nat Biotechnol 20:1041–1044

    Article  PubMed  CAS  Google Scholar 

  • Shinomura T, Nagatani A, Hanzawa H, Kubota M, Watanabe M, Furuya M (1996) Action spectrum for phytochrome-A and B-specific photoinduction of seed germination in Arabidopsis thaliana. Proc Natl Acad Sci USA 93:8129–8133

    Article  PubMed  CAS  Google Scholar 

  • Shinomura T, Uchida K, Furuya M (2000) Elementary processes of photoperception by phytochrome A for high irradiance response of hypocotyl elongation in Arabidopsis thaliana. Plant Physiol 122:147–156

    Article  PubMed  CAS  Google Scholar 

  • Stockhaus J, Nagatani A, Halfter U, Kay S, Furuya M, Chua N-H (1992) Serine-to-alanine substitutions at the amino-terminal region of phytochrome A result in an increase in biological activity. Genes Dev 6:2364–2372

    PubMed  CAS  Google Scholar 

  • Tepperman JM, Zhu T, Chang H-S, Wang X, Quail PH (2001) Multiple transcription-factor genes are early targets of phytochrome A signaling. Proc Natl Acad Sci USA 98:9437–9442

    Article  PubMed  CAS  Google Scholar 

  • Wagner JR, Brunzelle JS, Forest KT, Vierstra RD (2005) A light-sensing knot revealed by the structure of the chromophore-binding domain of phytochrome. Nature 438:325–331

    Article  PubMed  CAS  Google Scholar 

  • Whitelam GC, Johnson E, Peng J, Carol P, Anderson ML, Cowl JS, Harberd NP (1993) Phytochrome A null mutants of Arabidopsis display a wild-type phenotype in white light. Plant Cell 5:757–768

    Article  PubMed  CAS  Google Scholar 

  • Yanovsky MJ, Casal JJ, Luppi JP (1997) The VLF loci, polymorphic between ecotypes Landsberg erecta and Columbia dissect two branches of phytochrome A signalling pathways that correspond to the very-low fluence and high-irradiance responses of phytochrome. Plant J 12:659–667

    Article  PubMed  CAS  Google Scholar 

  • Yanovsky MJ, Whitelam GC, Casal JJ (2000) fhy3-1 retains inductive responses of phytochrome A. Plant Physiol 123:235–242

    Article  PubMed  CAS  Google Scholar 

  • Yanovsky MJ, Luppi JP, Kirchbauer D, Ogorodnikova OB, Sineshchekov VA, Adam E, Kircher S, Staneloni RJ, Schäfer E, Nagy F, Casal JJ (2002) Missense mutation in the PAS2 domain of phytochrome A impairs subnuclear localization and a subset of responses. Plant Cell 14:1591–1603

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Eberhard Schäfer (University of Freiburg) for allowing us to do the yeast two-hybrid experiments in his lab. This work was supported by the National Agency for the promotion of Science and Technology of Argentina (ANPCYT, grant BID 1728/OC-AR PICT 11631 to JJC), by University of Buenos Aires (grant G021 to JJC), by the Swiss National Science Foundation (grant PP00A−103005 to CF), the Human Frontier Science Program (HFSP) (grant RGY0016/2004-C to CF) and the University of Lausanne (to CF); by a fellowship from the HFSP to AH (LT00631/2003-C).

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Authors and Affiliations

  1. IFEVA, Facultad de Agronomía, Universidad de Buenos Aires and CONICET, Av. San Martín 4453, 1417, Buenos Aires, Argentina

    Santiago A. Trupkin & Jorge J. Casal

  2. Center for Integrative Genomics, University of Lausanne, Genopode Building, 1015, Lausanne, Switzerland

    Dimitry Debrieux & Christian Fankhauser

  3. Institute of Biology II/Botany, University of Freiburg, Schaenzlestrasse 1, 79104, Freiburg, Germany

    Andreas Hiltbrunner

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  1. Santiago A. Trupkin
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  2. Dimitry Debrieux
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  3. Andreas Hiltbrunner
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  4. Christian Fankhauser
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  5. Jorge J. Casal
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Correspondence to Jorge J. Casal.

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Trupkin, S.A., Debrieux, D., Hiltbrunner, A. et al. The serine-rich N-terminal region of Arabidopsis phytochrome A is required for protein stability. Plant Mol Biol 63, 669–678 (2007). https://doi.org/10.1007/s11103-006-9115-x

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  • DOI: https://doi.org/10.1007/s11103-006-9115-x

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