AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells

Matouš Glanc; Kasper Van Gelderen; Lukas Hoermayer; Shutang Tan; Satoshi Naramoto; Xixi Zhang; David Domjan; Ludmila Včelařová; Robert Hauschild; Alexander Johnson; Edward de Koning; Maritza van Dop; Eike Rademacher; Stef Janson; Xiaoyu Wei; Gergely Molnár; Matyáš Fendrych; Bert De Rybel; Remko Offringa; Jiří Friml

doi:10.1016/j.cub.2021.02.028

AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells

Matouš Glanc, Kasper Van Gelderen, Lukas Hoermayer, Shutang Tan, Satoshi Naramoto, Xixi Zhang, David Domjan, Ludmila Včelařová, Robert Hauschild, Alexander Johnson, Edward de Koning, Maritza van Dop, Eike Rademacher, Stef Janson, Xiaoyu Wei, Gergely Molnár, Matyáš Fendrych, Bert De Rybel, Remko Offringa, Jiří Friml

Sub Plant Ecophysiology

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Polar subcellular localization of the PIN exporters of the phytohormone auxin is a key determinant of directional, intercellular auxin transport and thus a central topic of both plant cell and developmental biology. Arabidopsis mutants lacking PID, a kinase that phosphorylates PINs, or the MAB4/MEL proteins of unknown molecular function display PIN polarity defects and phenocopy pin mutants, but mechanistic insights into how these factors convey PIN polarity are missing. Here, by combining protein biochemistry with quantitative live-cell imaging, we demonstrate that PINs, MAB4/MELs, and AGC kinases interact in the same complex at the plasma membrane. MAB4/MELs are recruited to the plasma membrane by the PINs and in concert with the AGC kinases maintain PIN polarity through limiting lateral diffusion-based escape of PINs from the polar domain. The PIN-MAB4/MEL-PID protein complex has self-reinforcing properties thanks to positive feedback between AGC kinase-mediated PIN phosphorylation and MAB4/MEL recruitment. We thus uncover the molecular mechanism by which AGC kinases and MAB4/MEL proteins regulate PIN localization and plant development.

Original language	English
Pages (from-to)	1918-1930.e5
Number of pages	18
Journal	Current Biology
Volume	31
Issue number	9
Early online date	10 Mar 2021
DOIs	https://doi.org/10.1016/j.cub.2021.02.028
Publication status	Published - 10 May 2021

Bibliographical note

Funding Information:
We acknowledge Ben Scheres, Christian Luschnig, and Claus Schwechheimer for sharing published material. We thank Monika Hrtyan and Dorota Jaworska at IST Austria and Gerda Lamers and Ward de Winter at IBL Netherlands for technical assistance; Corinna Hartinger, Jakub Hajn?, Lesia Rodriguez, Mingyue Li, and Lindy Abas for experimental support; and the Bioimaging Facility at IST Austria and the Bioimaging Core at VIB for imaging support. We are grateful to Christian Luschnig, Lindy Abas, and Roman Pleskot for valuable discussions. We also acknowledge the EMBO for supporting M.G. with a long-term fellowship (ALTF 1005-2019) during the finalization and revision of this manuscript in the laboratory of B.D.R. and we thank R. Pierik for allowing K.V.G. to work on this manuscript during a postdoc in his laboratory at Utrecht University. This work was supported by grants from the European Research Council under the European Union's Seventh Framework Programme (ERC grant agreements 742985 to J.F. 714055 to B.D.R. and 803048 to M.F.), the Austrian Science Fund (FWF; I 3630-B25 to J.F.), Chemical Sciences (partly) financed by the Dutch Research Council (NWO-CW TOP 700.58.301 to R.O.), the Dutch Research Council (NWO-VICI 865.17.002 to R. Pierik), Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (KAKENHI grant 17K17595 to S.N.), the Ministry of Education, Youth and Sports of the Czech Republic (M?MT project NPUI-LO1417), and a China Scholarship Council (to X.W.). M.G. and K.V.G. conceptualized the study, and designed, performed, and analyzed the majority of experiments with equal contributions; L.H. S.T. S.N. X.Z. D.D. L.V. R.H. A.J. E.d.K. M.v.D. E.R. S.J. and X.W. contributed to performing and analyzing experiments; E.R. M.F. and B.D.R. contributed to conceptualizing the study; R.O. and J.F. acquired funding, conceptualized the study, supervised the work, and analyzed experiments with equal contributions; M.G. K.V.G. R.O. and J.F. wrote the manuscript with substantial input from S.T. M.F. and B.D.R.; and all authors have had the chance to read and comment on the manuscript. The authors declare no competing interests.

Funding Information:
We acknowledge Ben Scheres, Christian Luschnig, and Claus Schwechheimer for sharing published material. We thank Monika Hrtyan and Dorota Jaworska at IST Austria and Gerda Lamers and Ward de Winter at IBL Netherlands for technical assistance; Corinna Hartinger, Jakub Hajný, Lesia Rodriguez, Mingyue Li, and Lindy Abas for experimental support; and the Bioimaging Facility at IST Austria and the Bioimaging Core at VIB for imaging support. We are grateful to Christian Luschnig, Lindy Abas, and Roman Pleskot for valuable discussions. We also acknowledge the EMBO for supporting M.G. with a long-term fellowship ( ALTF 1005-2019 ) during the finalization and revision of this manuscript in the laboratory of B.D.R., and we thank R. Pierik for allowing K.V.G. to work on this manuscript during a postdoc in his laboratory at Utrecht University. This work was supported by grants from the European Research Council under the European Union’s Seventh Framework Programme (ERC grant agreements 742985 to J.F., 714055 to B.D.R., and 803048 to M.F.), the Austrian Science Fund (FWF; I 3630-B25 to J.F.), Chemical Sciences (partly) financed by the Dutch Research Council (NWO-CW TOP 700.58.301 to R.O.), the Dutch Research Council (NWO-VICI 865.17.002 to R. Pierik), Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (KAKENHI grant 17K17595 to S.N.), the Ministry of Education, Youth and Sports of the Czech Republic (MŠMT project NPUI-LO1417 ), and a China Scholarship Council (to X.W.).

Publisher Copyright:
© 2021 The Authors

Keywords

Arabidopsis
cell polarity
lateral diffusion
plant development
polar auxin transport
positive feedback
protein phosphorylation

Access to Document

10.1016/j.cub.2021.02.028Licence: CC BY

PIIS0960982221002694Final published version, 4.24 MBLicence: CC BY

Cite this

Glanc, M., Van Gelderen, K., Hoermayer, L., Tan, S., Naramoto, S., Zhang, X., Domjan, D., Včelařová, L., Hauschild, R., Johnson, A., de Koning, E., van Dop, M., Rademacher, E., Janson, S., Wei, X., Molnár, G., Fendrych, M., De Rybel, B., Offringa, R., & Friml, J. (2021). AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells. Current Biology, 31(9), 1918-1930.e5. https://doi.org/10.1016/j.cub.2021.02.028

@article{62692c738af74e6684315c29951784f1,

title = "AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells",

abstract = "Polar subcellular localization of the PIN exporters of the phytohormone auxin is a key determinant of directional, intercellular auxin transport and thus a central topic of both plant cell and developmental biology. Arabidopsis mutants lacking PID, a kinase that phosphorylates PINs, or the MAB4/MEL proteins of unknown molecular function display PIN polarity defects and phenocopy pin mutants, but mechanistic insights into how these factors convey PIN polarity are missing. Here, by combining protein biochemistry with quantitative live-cell imaging, we demonstrate that PINs, MAB4/MELs, and AGC kinases interact in the same complex at the plasma membrane. MAB4/MELs are recruited to the plasma membrane by the PINs and in concert with the AGC kinases maintain PIN polarity through limiting lateral diffusion-based escape of PINs from the polar domain. The PIN-MAB4/MEL-PID protein complex has self-reinforcing properties thanks to positive feedback between AGC kinase-mediated PIN phosphorylation and MAB4/MEL recruitment. We thus uncover the molecular mechanism by which AGC kinases and MAB4/MEL proteins regulate PIN localization and plant development.",

keywords = "Arabidopsis, cell polarity, lateral diffusion, plant development, polar auxin transport, positive feedback, protein phosphorylation",

author = "Matou{\v s} Glanc and {Van Gelderen}, Kasper and Lukas Hoermayer and Shutang Tan and Satoshi Naramoto and Xixi Zhang and David Domjan and Ludmila V{\v c}ela{\v r}ov{\'a} and Robert Hauschild and Alexander Johnson and {de Koning}, Edward and {van Dop}, Maritza and Eike Rademacher and Stef Janson and Xiaoyu Wei and Gergely Moln{\'a}r and Maty{\'a}{\v s} Fendrych and {De Rybel}, Bert and Remko Offringa and Ji{\v r}{\'i} Friml",

note = "Funding Information: We acknowledge Ben Scheres, Christian Luschnig, and Claus Schwechheimer for sharing published material. We thank Monika Hrtyan and Dorota Jaworska at IST Austria and Gerda Lamers and Ward de Winter at IBL Netherlands for technical assistance; Corinna Hartinger, Jakub Hajn?, Lesia Rodriguez, Mingyue Li, and Lindy Abas for experimental support; and the Bioimaging Facility at IST Austria and the Bioimaging Core at VIB for imaging support. We are grateful to Christian Luschnig, Lindy Abas, and Roman Pleskot for valuable discussions. We also acknowledge the EMBO for supporting M.G. with a long-term fellowship (ALTF 1005-2019) during the finalization and revision of this manuscript in the laboratory of B.D.R. and we thank R. Pierik for allowing K.V.G. to work on this manuscript during a postdoc in his laboratory at Utrecht University. This work was supported by grants from the European Research Council under the European Union's Seventh Framework Programme (ERC grant agreements 742985 to J.F. 714055 to B.D.R. and 803048 to M.F.), the Austrian Science Fund (FWF; I 3630-B25 to J.F.), Chemical Sciences (partly) financed by the Dutch Research Council (NWO-CW TOP 700.58.301 to R.O.), the Dutch Research Council (NWO-VICI 865.17.002 to R. Pierik), Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (KAKENHI grant 17K17595 to S.N.), the Ministry of Education, Youth and Sports of the Czech Republic (M?MT project NPUI-LO1417), and a China Scholarship Council (to X.W.). M.G. and K.V.G. conceptualized the study, and designed, performed, and analyzed the majority of experiments with equal contributions; L.H. S.T. S.N. X.Z. D.D. L.V. R.H. A.J. E.d.K. M.v.D. E.R. S.J. and X.W. contributed to performing and analyzing experiments; E.R. M.F. and B.D.R. contributed to conceptualizing the study; R.O. and J.F. acquired funding, conceptualized the study, supervised the work, and analyzed experiments with equal contributions; M.G. K.V.G. R.O. and J.F. wrote the manuscript with substantial input from S.T. M.F. and B.D.R.; and all authors have had the chance to read and comment on the manuscript. The authors declare no competing interests. Funding Information: We acknowledge Ben Scheres, Christian Luschnig, and Claus Schwechheimer for sharing published material. We thank Monika Hrtyan and Dorota Jaworska at IST Austria and Gerda Lamers and Ward de Winter at IBL Netherlands for technical assistance; Corinna Hartinger, Jakub Hajn{\'y}, Lesia Rodriguez, Mingyue Li, and Lindy Abas for experimental support; and the Bioimaging Facility at IST Austria and the Bioimaging Core at VIB for imaging support. We are grateful to Christian Luschnig, Lindy Abas, and Roman Pleskot for valuable discussions. We also acknowledge the EMBO for supporting M.G. with a long-term fellowship ( ALTF 1005-2019 ) during the finalization and revision of this manuscript in the laboratory of B.D.R., and we thank R. Pierik for allowing K.V.G. to work on this manuscript during a postdoc in his laboratory at Utrecht University. This work was supported by grants from the European Research Council under the European Union{\textquoteright}s Seventh Framework Programme (ERC grant agreements 742985 to J.F., 714055 to B.D.R., and 803048 to M.F.), the Austrian Science Fund (FWF; I 3630-B25 to J.F.), Chemical Sciences (partly) financed by the Dutch Research Council (NWO-CW TOP 700.58.301 to R.O.), the Dutch Research Council (NWO-VICI 865.17.002 to R. Pierik), Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (KAKENHI grant 17K17595 to S.N.), the Ministry of Education, Youth and Sports of the Czech Republic (M{\v S}MT project NPUI-LO1417 ), and a China Scholarship Council (to X.W.). Publisher Copyright: {\textcopyright} 2021 The Authors",

year = "2021",

month = may,

day = "10",

doi = "10.1016/j.cub.2021.02.028",

language = "English",

volume = "31",

pages = "1918--1930.e5",

journal = "Current Biology",

issn = "0960-9822",

publisher = "Cell Press",

number = "9",

}

Glanc, M, Van Gelderen, K, Hoermayer, L, Tan, S, Naramoto, S, Zhang, X, Domjan, D, Včelařová, L, Hauschild, R, Johnson, A, de Koning, E, van Dop, M, Rademacher, E, Janson, S, Wei, X, Molnár, G, Fendrych, M, De Rybel, B, Offringa, R & Friml, J 2021, 'AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells', Current Biology, vol. 31, no. 9, pp. 1918-1930.e5. https://doi.org/10.1016/j.cub.2021.02.028

TY - JOUR

T1 - AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells

AU - Glanc, Matouš

AU - Van Gelderen, Kasper

AU - Hoermayer, Lukas

AU - Tan, Shutang

AU - Naramoto, Satoshi

AU - Zhang, Xixi

AU - Domjan, David

AU - Včelařová, Ludmila

AU - Hauschild, Robert

AU - Johnson, Alexander

AU - de Koning, Edward

AU - van Dop, Maritza

AU - Rademacher, Eike

AU - Janson, Stef

AU - Wei, Xiaoyu

AU - Molnár, Gergely

AU - Fendrych, Matyáš

AU - De Rybel, Bert

AU - Offringa, Remko

AU - Friml, Jiří

N1 - Funding Information: We acknowledge Ben Scheres, Christian Luschnig, and Claus Schwechheimer for sharing published material. We thank Monika Hrtyan and Dorota Jaworska at IST Austria and Gerda Lamers and Ward de Winter at IBL Netherlands for technical assistance; Corinna Hartinger, Jakub Hajn?, Lesia Rodriguez, Mingyue Li, and Lindy Abas for experimental support; and the Bioimaging Facility at IST Austria and the Bioimaging Core at VIB for imaging support. We are grateful to Christian Luschnig, Lindy Abas, and Roman Pleskot for valuable discussions. We also acknowledge the EMBO for supporting M.G. with a long-term fellowship (ALTF 1005-2019) during the finalization and revision of this manuscript in the laboratory of B.D.R. and we thank R. Pierik for allowing K.V.G. to work on this manuscript during a postdoc in his laboratory at Utrecht University. This work was supported by grants from the European Research Council under the European Union's Seventh Framework Programme (ERC grant agreements 742985 to J.F. 714055 to B.D.R. and 803048 to M.F.), the Austrian Science Fund (FWF; I 3630-B25 to J.F.), Chemical Sciences (partly) financed by the Dutch Research Council (NWO-CW TOP 700.58.301 to R.O.), the Dutch Research Council (NWO-VICI 865.17.002 to R. Pierik), Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (KAKENHI grant 17K17595 to S.N.), the Ministry of Education, Youth and Sports of the Czech Republic (M?MT project NPUI-LO1417), and a China Scholarship Council (to X.W.). M.G. and K.V.G. conceptualized the study, and designed, performed, and analyzed the majority of experiments with equal contributions; L.H. S.T. S.N. X.Z. D.D. L.V. R.H. A.J. E.d.K. M.v.D. E.R. S.J. and X.W. contributed to performing and analyzing experiments; E.R. M.F. and B.D.R. contributed to conceptualizing the study; R.O. and J.F. acquired funding, conceptualized the study, supervised the work, and analyzed experiments with equal contributions; M.G. K.V.G. R.O. and J.F. wrote the manuscript with substantial input from S.T. M.F. and B.D.R.; and all authors have had the chance to read and comment on the manuscript. The authors declare no competing interests. Funding Information: We acknowledge Ben Scheres, Christian Luschnig, and Claus Schwechheimer for sharing published material. We thank Monika Hrtyan and Dorota Jaworska at IST Austria and Gerda Lamers and Ward de Winter at IBL Netherlands for technical assistance; Corinna Hartinger, Jakub Hajný, Lesia Rodriguez, Mingyue Li, and Lindy Abas for experimental support; and the Bioimaging Facility at IST Austria and the Bioimaging Core at VIB for imaging support. We are grateful to Christian Luschnig, Lindy Abas, and Roman Pleskot for valuable discussions. We also acknowledge the EMBO for supporting M.G. with a long-term fellowship ( ALTF 1005-2019 ) during the finalization and revision of this manuscript in the laboratory of B.D.R., and we thank R. Pierik for allowing K.V.G. to work on this manuscript during a postdoc in his laboratory at Utrecht University. This work was supported by grants from the European Research Council under the European Union’s Seventh Framework Programme (ERC grant agreements 742985 to J.F., 714055 to B.D.R., and 803048 to M.F.), the Austrian Science Fund (FWF; I 3630-B25 to J.F.), Chemical Sciences (partly) financed by the Dutch Research Council (NWO-CW TOP 700.58.301 to R.O.), the Dutch Research Council (NWO-VICI 865.17.002 to R. Pierik), Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology, Japan (KAKENHI grant 17K17595 to S.N.), the Ministry of Education, Youth and Sports of the Czech Republic (MŠMT project NPUI-LO1417 ), and a China Scholarship Council (to X.W.). Publisher Copyright: © 2021 The Authors

PY - 2021/5/10

Y1 - 2021/5/10

N2 - Polar subcellular localization of the PIN exporters of the phytohormone auxin is a key determinant of directional, intercellular auxin transport and thus a central topic of both plant cell and developmental biology. Arabidopsis mutants lacking PID, a kinase that phosphorylates PINs, or the MAB4/MEL proteins of unknown molecular function display PIN polarity defects and phenocopy pin mutants, but mechanistic insights into how these factors convey PIN polarity are missing. Here, by combining protein biochemistry with quantitative live-cell imaging, we demonstrate that PINs, MAB4/MELs, and AGC kinases interact in the same complex at the plasma membrane. MAB4/MELs are recruited to the plasma membrane by the PINs and in concert with the AGC kinases maintain PIN polarity through limiting lateral diffusion-based escape of PINs from the polar domain. The PIN-MAB4/MEL-PID protein complex has self-reinforcing properties thanks to positive feedback between AGC kinase-mediated PIN phosphorylation and MAB4/MEL recruitment. We thus uncover the molecular mechanism by which AGC kinases and MAB4/MEL proteins regulate PIN localization and plant development.

AB - Polar subcellular localization of the PIN exporters of the phytohormone auxin is a key determinant of directional, intercellular auxin transport and thus a central topic of both plant cell and developmental biology. Arabidopsis mutants lacking PID, a kinase that phosphorylates PINs, or the MAB4/MEL proteins of unknown molecular function display PIN polarity defects and phenocopy pin mutants, but mechanistic insights into how these factors convey PIN polarity are missing. Here, by combining protein biochemistry with quantitative live-cell imaging, we demonstrate that PINs, MAB4/MELs, and AGC kinases interact in the same complex at the plasma membrane. MAB4/MELs are recruited to the plasma membrane by the PINs and in concert with the AGC kinases maintain PIN polarity through limiting lateral diffusion-based escape of PINs from the polar domain. The PIN-MAB4/MEL-PID protein complex has self-reinforcing properties thanks to positive feedback between AGC kinase-mediated PIN phosphorylation and MAB4/MEL recruitment. We thus uncover the molecular mechanism by which AGC kinases and MAB4/MEL proteins regulate PIN localization and plant development.

KW - Arabidopsis

KW - cell polarity

KW - lateral diffusion

KW - plant development

KW - polar auxin transport

KW - positive feedback

KW - protein phosphorylation

U2 - 10.1016/j.cub.2021.02.028

DO - 10.1016/j.cub.2021.02.028

M3 - Article

C2 - 33705718

SN - 0960-9822

VL - 31

SP - 1918-1930.e5

JO - Current Biology

JF - Current Biology

IS - 9

ER -

AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells

Abstract

Bibliographical note

Keywords

Access to Document

Fingerprint

Cite this