Abstract
Mitogen-activated protein kinases (MPKs) play a crucial role in the regulation of many aspects of plant growth, development, and responses to the environment. In Arabidopsis, MPK3 and MPK6 are known to be implicated in embryonic and stomatal development as well as biotic and abiotic stress responses. However, the regulation mechanisms of plant developments by MPKs are poorly understood. Here, we provide direct evidences that ASYMMETRIC LEAVES1 (AS1), a MYB transcription factor, is phosphorylated by AtMPK3 and AtMPK6. We also show the direct interactions between AS1 and AtMPK3/6 by using yeast and plant systems. In addition, we identified the phosphorylation of AS1 by recombinant AtMPK3 and AtMPK6 by in vitro kinase assays. Using site-directed mutagenesis assay, we identified that the 202th serine residue of AS1 was specifically phosphoylated by MPK3/6. Finally, we confirmed that AS1 could be phosphorylated by endogenous AtMPK3/6 using in-gel kinase assays. These results suggest that the phosphorylation of AS1 by MPKs may be one of regulation modes for leaf development in plants.
Similar content being viewed by others
References
Biondi RM, Nebreda AR (2003) Signaling specificity of Ser/Thr protein kinases through docking-site-mediated interactions. Biochem J 372:1–13
Bögre L, Calderini O, Binarova P, Mattauch M, Till S, Kiegerl S, Jonak C, Pollaschek C, Barker P, Huskisson NS, Hirt H, Heberle-Bore E (1999) A MAP kinase is activated late in plant mitosis and becomes localized to the plane of cell division. Plant Cell 11:101–113
Bögre L, Meskiene I, Heberle-Bors E, Hirt H (2000) Stressing the role of MAP kinases in mitogenic stimulation. Plant Mol Biol 43:705–718
Bowman JL, Eshed Y (2000) Formation and maintenance of the shoot apical meristem. Trends Plant Sci 5:110–115
Bush SM, Krysan PJ (2007) Mutational evidence that the Arabidopsis MAP kinase MPK6 is involved in anther, inflorescence, and embryo development. J Exp Bot 58:2181–2191
Byrne ME, Barley R, Curtis M, Arroyo JM, Dunham M, Hudson A, Martienssen RA (2000) Asymmetric leaves1 mediates leaf patterning and stem cell function in Arabidopsis. Nature 408:967–971
Calderini O, Bögre L, Vicente O, Binarova P, Heberle-Bors E, Wilson C (1998) A cell cycle regulated MAP kinase with a possible role in cytokinesis in tobacco cells. J Cell Sci 111:3091–3100
Chen H, Zou Y, Shang Y, Lin H, Wang Y, Cai R, Tang X, Zhou JM (2008) Firefly luciferase complementation imaging assay for protein-protein interactions in plants. Plant Physiol 146:368–376
Desikan R, Hancock JT, Ichimura K, Shinozaki K, Neill SJ (2001) Harpin induces activation of the Arabidopsis mitogen-activated protein kinases AtMPK4 and AtMPK6. Plant Physiol 126:1579–1587
Guo M, Thomas J, Collins G, Timmermans MC (2008) Direct repression of KNOX loci by the ASYMMETRIC LEAVES1 complex of Arabidopsis. Plant Cell 20:48–58
Ichimura K et al (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends Plant Sci 7:301–308
Ichimura K, Mizoguchi T, Yoshida R, Yuasa T, Shinozaki K (2000) Various abiotic stresses rapidly activate Arabidopsis MAP kinases ATMPK4 and ATMPK6. Plant J 24:655–665
Ikezaki M, Kojima M, Sakakibara H, Kojima S, Ueno Y, Machida C, Machida Y (2010) Genetic networks regulated by ASYMMETRIC LEAVES1 (AS1) and AS2 in leaf development in Arabidopsis thaliana: KNOX genes control five morphological events. Plant J 61:70–82
Iwakawa H, Iwasaki M, Kojima S, Ueno Y, Soma T, Tanaka H, Semiarti E, Machida Y, Machida C (2007) Expression of the ASYMMETRIC LEAVES2 gene in the adaxial domain of Arabidopsis leaves represses cell proliferation in this domain and is critical for the development of properly expanded leaves. Plant J 51:173–184
Jonak C, Okrész L, Bgre L, Hirt H (2002) Complexity, cross talk and integration of plant MAP kinase signalling. Curr Opin Plant Biol 5:415–424
Lampard GR, Macalister CA, Bergmann DC (2008) Arabidopsis stomatal initiation is controlled by MAPK-mediated regulation of the bHLH SPEECHLESS. Science 322:1113–1116
Li H, Xu L, Wang H, Yuan Z, Cao X, Yang Z, Zhang D, Xu Y, Huang H (2005) The putative RNA-dependent RNA polymerase RDR6 acts synergistically with ASYMMETRIC LEAVES1 and 2 to repress BREVIPEDICELLUS and MicroRNA165/166 in Arabidopsis leaf development. Plant Cell 17:2157–2171
Mishra NS, Tuteja R, Tuteja N (2006) Signaling through MAP kinase networks in plants. Arch Biochem Biophys 452:55–68
Mizoguchi T, Irie K, Hirayama T, Hayashida N, Yamaguchi-Shinozaki K, Matsumoto K, Shinozaki K (1996) A gene encoding a mitogenactivated protein kinase kinase kinase is induced simultaneously with genes for a mitogen-activated protein kinase and an S6 ribosomal protein kinase by touch, cold, and water stress in Arabidopsis thaliana. Proc Natl Acad Sci USA 93:765–769
Morse AM, Whetten RW, Dubos C, Campbell MM (2009) Posttranslational modification of an R2R3-MYB transcription factor by a MAP Kinase during xylem development. New Phytol 183:1001–1013
Moyano E, MartÍnez-Garcia JF, Martin C (1996) Apparent redundancy in myb gene function provides gearing for the control of flavonoid biosynthesis in antirrhinum flowers. Plant Cell 8:1519–1532
Nühse TS, Peck SC, Hirt H, Boller T (2000) Microbial elicitors induce activation and dual phosphorylation of the Arabidopsis thaliana MPK6. J Biol Chem 275:7521–7526
Nurmberg PL, Knox KA, Yun BW, Morris PC, Shafiei R, Hudson A, Loake GL (2007) The developmental selector AS1 is an evolutionarily conserved regulator of the plant immune response. Proc Natl Acad Sci USA 104:19795–18800
Ori N, Eshed Y, Chuck G, Bowman JL, Hake S (2000) Mechanisms that control knox gene expression in the Arabidopsis shoot. Development 127:5523–5532
Park JM, Paek KH (2007) Recognition and response in plantpathogen interactions. J Plant Biol 50:132–138
Phelps-Durr TL, Thomas J, Vahab P, Timmermans MCP (2005) Maize rough sheath2 and its Arabidopsis orthologue ASYMMETRIC LEAVES1 interact with HIRA, a predicted histone chaperone, to maintain knox gene silencing and determinacy during organogenesis. Plant Cell 17:2886–2898
Rodriguez MC, Petersen M, Mundy J (2010) Mitogen-activated protein kinase signaling in plants. Annu Rev Plant Biol 61:621–649
Semiarti E, Ueno Y, Tsukaya H, Iwakawa H, Machida C, Machida Y (2001) The ASYMMETRIC LEAVES2 gene of Arabidopsis thaliana regulates formation of a symmetric lamina, establishment of venation and repression of meristem-related homeobox genes in leaves. Development 128:1771–1783
Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456
Szakonyi D, Moschopoulos A, Byrne ME (2010) Perspectives on leaf dorsoventral polarity. J Plant Res 123:281–290
Takenaka K, Moriguchi T, Nishida E (1998) Activation of the protein kinase p38 in the spindle assembly checkpoint and mitotic arrest. Science 280:599–602.
Timmermans MCP, Hudson A, Becraft PW, Nelson T (1999) ROUGH SHEATH2: a Myb protein that represses knox homeo-box genes in maize lateral organ primordia. Science 284:151–153
Tootle TL, Rebay I (2005) Post-translational modifications influence transcription factor activity: a view from the ETS superfamily. Bioessays 27:285–298
Uchida N, Kimura S, Koenig D, Sinha N (2010) Coordination of leaf development via regulation of KNOX1 genes. J Plant Res 123:7–14
Verlhac MH, Kubiak JZ, Clarke HJ, Maro B (1994) Microtubule and chromatin behavior follow MAP kinase activity but not MPF activity during meiosis in mouse oocytes. Development 120:1017–1025
Waites R, Hudson A (1995) Phantastica: a gene required for dorsoventrality of leaves in Antirrhinum majus. Development 121:2143–2154
Wang H, Liu Y, Bruffett K, Lee J, Hause G, Walker JC, Zhang S (2008) Haplo-insufficiency of MPK3 in MPK6 mutant background uncovers a novel function of these two MAPKs in Arabidopsis ovule development. Plant Cell 20:602–613
Wang H, Ngwenyama N, Liu Y, Walker JC, Zhang S (2007) Stomatal development and patterning are regulated by environmentally responsive mitogen-activated protein kinases in Arabidopsis. Plant Cell 19:63–73
Xu L, Xu Y, Dong A, Sun Y, Pi L, Xu Y, Huang H (2003) Novel as1 and as2 defects in leaf adaxial-abaxial polarity reveal the requirement for ASYMMETRIC LEAVES1 and 2 and ERECTA functions in specifying leaf adaxial identity. Development 130:4097–4107
Yang SH, Sharrocks AD, Whitmarsh AJ (2003) Transcriptional regulation by the MAP kinase signaling cascades. Gene 320:3–21
Zhang S, Jin CD, Roux SJ (1993) Casein kinase II-type protein kinase from pea cytoplasm and its inactivation by alkaline phosphatase in vitro. Plant Physiol 103:955–962
Zhang Y, Dong C (2007) Regulatory mechanisms of mitogen-activated kinase signaling. Cell Mol Life Sci 64:2771–2789
Author information
Authors and Affiliations
Corresponding authors
Additional information
These authors contributed equally to this work.
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Park, H.C., Han, H.J., Lee, S.M. et al. ASYMMETRIC LEAVES1 is phosphorylated by MPK3/6 in Arabidopsis thaliana . J. Plant Biol. 56, 208–215 (2013). https://doi.org/10.1007/s12374-013-0123-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12374-013-0123-5