WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop

Megan J. Agajanian; Matthew P. Walker; Alison D. Axtman; Roberta R. Ruela-de-Sousa; D. Stephen Serafin; Alex D. Rabinowitz; David M. Graham; Meagan B. Ryan; Tigist Tamir; Yuko Nakamichi; Melissa V. Gammons; James M. Bennett; Rafael M. Couñago; David H. Drewry; Jonathan M. Elkins; Carina Gileadi; Opher Gileadi; Paulo H. Godoi; Nirav Kapadia; Susanne Müller; André S. Santiago; Fiona J. Sorrell; Carrow I. Wells; Oleg Fedorov; Timothy M. Willson; William J. Zuercher; Michael B. Major

doi:10.1016/j.celrep.2018.12.023

WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop

Megan J. Agajanian, Matthew P. Walker, Alison D. Axtman, Roberta R. Ruela-de-Sousa, D. Stephen Serafin, Alex D. Rabinowitz, David M. Graham, Meagan B. Ryan, Tigist Tamir, Yuko Nakamichi, Melissa V. Gammons, James M. Bennett, Rafael M. Couñago, David H. Drewry, Jonathan M. Elkins, Carina Gileadi, Opher Gileadi, Paulo H. Godoi, Nirav Kapadia, Susanne MüllerAndré S. Santiago, Fiona J. Sorrell, Carrow I. Wells, Oleg Fedorov, Timothy M. Willson, William J. Zuercher, Michael B. Major

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

41 Scopus citations

Abstract

WNT signal transduction is essential for normal development and contributes to many human diseases. Agajanian et al. used a kinase gain-of-function screen to show that WNT activates the AAK1 kinase to promote clathrin-mediated endocytosis of the WNT receptor. This work identifies an AAK-driven negative feedback loop that downregulates WNT signaling.

Original language	English
Pages (from-to)	79-93.e8
Journal	Cell Reports
Volume	26
Issue number	1
DOIs	https://doi.org/10.1016/j.celrep.2018.12.023
State	Published - Jan 2 2019

Keywords

AAK1
AP2M1
LRP6
WNT signaling
clathrin
endocytosis
feedback loop
gain-of-function screen
kinase

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10.1016/j.celrep.2018.12.023

Cite this

Agajanian, M. J., Walker, M. P., Axtman, A. D., Ruela-de-Sousa, R. R., Serafin, D. S., Rabinowitz, A. D., Graham, D. M., Ryan, M. B., Tamir, T., Nakamichi, Y., Gammons, M. V., Bennett, J. M., Couñago, R. M., Drewry, D. H., Elkins, J. M., Gileadi, C., Gileadi, O., Godoi, P. H., Kapadia, N., ... Major, M. B. (2019). WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop. Cell Reports, 26(1), 79-93.e8. https://doi.org/10.1016/j.celrep.2018.12.023

@article{98d69f7871004844919b79f2340190f7,

title = "WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop",

abstract = "WNT signal transduction is essential for normal development and contributes to many human diseases. Agajanian et al. used a kinase gain-of-function screen to show that WNT activates the AAK1 kinase to promote clathrin-mediated endocytosis of the WNT receptor. This work identifies an AAK-driven negative feedback loop that downregulates WNT signaling.",

keywords = "AAK1, AP2M1, LRP6, WNT signaling, clathrin, endocytosis, feedback loop, gain-of-function screen, kinase",

author = "Agajanian, {Megan J.} and Walker, {Matthew P.} and Axtman, {Alison D.} and Ruela-de-Sousa, {Roberta R.} and Serafin, {D. Stephen} and Rabinowitz, {Alex D.} and Graham, {David M.} and Ryan, {Meagan B.} and Tigist Tamir and Yuko Nakamichi and Gammons, {Melissa V.} and Bennett, {James M.} and Cou{\~n}ago, {Rafael M.} and Drewry, {David H.} and Elkins, {Jonathan M.} and Carina Gileadi and Opher Gileadi and Godoi, {Paulo H.} and Nirav Kapadia and Susanne M{\"u}ller and Santiago, {Andr{\'e} S.} and Sorrell, {Fiona J.} and Wells, {Carrow I.} and Oleg Fedorov and Willson, {Timothy M.} and Zuercher, {William J.} and Major, {Michael B.}",

note = "Funding Information: M.B.M. acknowledges support from the NIH ( RO1-CA187799 and U24-DK116204-01 ). M.J.A. received financial support from NIH T32 Predoctoral Training Grants in Pharmacology ( T32-GM007040-43 and T32-GM007040-42 ), an Initiative for Maximizing Student Diversity Grant ( R25-GM055336-16 ), and the NIH National Cancer Institute (NCI) NRSA Predoctoral Fellowship to Promote Diversity in Health-Related Research ( F31CA228289 ). M.P.W. received support from the Lymphoma Research Foundation ( 337444 ) and the NIH ( T32-CA009156-35 ). Y.N. was supported by grants-in-aid from the Japan Society for the Promotion of Science (JSPS) ( 15KK0356 and 16K11493 ). T.T. was supported by the Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study. M.V.G. was supported by Cancer Research UK (grants C7379/A15291 and C7379/A24639 to Mariann Bienz). The UNC Flow Cytometry Core Facility is supported in part by Cancer Center Core Support Grant P30 CA016086 to the UNC Lineberger Comprehensive Cancer Center, and research reported in this publication was supported by the Center for AIDS Research (award number 5P30AI050410 ), and the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie , Bayer Pharma AG , Boehringer Ingelheim , the Canada Foundation for Innovation , the Eshelman Institute for Innovation , Genome Canada , the Innovative Medicines Initiative ( European Union [EU]/European Federation of Pharmaceutical Industries and Associations [EFPIA]) (ULTRA-DD grant no. 115766), Janssen , Merck & Company , Merck KGaA , Novartis Pharma AG , the Ontario Ministry of Economic Development and Innovation , Pfizer , the S{\~a}o Paulo Research Foundation (FAPESP) ( 2013/50724-5 ), Takeda , and the Wellcome Trust ( 106169/ZZ14/Z ). R.R.R. received financial support from FAPESP ( 2016/17469-0 ). We would also like to thank Claire Strain-Damerell and Pavel Savitsky for cloning various mutants of AAK1 and BMP2K proteins that were used in the crystallization trials. Additionally, we thank Dr. Sean Conner for providing the AAK1 plasmids, Dr. Stephane Angers for kindly providing the HEK293T DVL TKO cells, and Dr. Mariann Bienz for providing comments and feedback. We would like to thank members of the Major laboratory for their feedback and expertise regarding experimental design and project direction. Funding Information: M.B.M. acknowledges support from the NIH (RO1-CA187799 and U24-DK116204-01). M.J.A. received financial support from NIH T32 Predoctoral Training Grants in Pharmacology (T32-GM007040-43 and T32-GM007040-42), an Initiative for Maximizing Student Diversity Grant (R25-GM055336-16), and the NIH National Cancer Institute (NCI) NRSA Predoctoral Fellowship to Promote Diversity in Health-Related Research (F31CA228289). M.P.W. received support from the Lymphoma Research Foundation (337444) and the NIH (T32-CA009156-35). Y.N. was supported by grants-in-aid from the Japan Society for the Promotion of Science (JSPS) (15KK0356 and 16K11493). T.T. was supported by the Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study. M.V.G. was supported by Cancer Research UK (grants C7379/A15291 and C7379/A24639 to Mariann Bienz). The UNC Flow Cytometry Core Facility is supported in part by Cancer Center Core Support Grant P30 CA016086 to the UNC Lineberger Comprehensive Cancer Center, and research reported in this publication was supported by the Center for AIDS Research (award number 5P30AI050410), and the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, the Canada Foundation for Innovation, the Eshelman Institute for Innovation, Genome Canada, the Innovative Medicines Initiative (European Union [EU]/European Federation of Pharmaceutical Industries and Associations [EFPIA]) (ULTRA-DD grant no. 115766), Janssen, Merck & Company, Merck KGaA, Novartis Pharma AG, the Ontario Ministry of Economic Development and Innovation, Pfizer, the S?o Paulo Research Foundation (FAPESP) (2013/50724-5), Takeda, and the Wellcome Trust (106169/ZZ14/Z). R.R.R. received financial support from FAPESP (2016/17469-0). We would also like to thank Claire Strain-Damerell and Pavel Savitsky for cloning various mutants of AAK1 and BMP2K proteins that were used in the crystallization trials. Additionally, we thank Dr. Sean Conner for providing the AAK1 plasmids, Dr. Stephane Angers for kindly providing the HEK293T DVL TKO cells, and Dr. Mariann Bienz for providing comments and feedback. We would like to thank members of the Major laboratory for their feedback and expertise regarding experimental design and project direction. Funding Information: M.B.M. acknowledges support from the NIH (RO1-CA187799 and U24-DK116204-01). M.J.A. received financial support from NIH T32 Predoctoral Training Grants in Pharmacology (T32-GM007040-43 and T32-GM007040-42), an Initiative for Maximizing Student Diversity Grant (R25-GM055336-16), and the NIH National Cancer Institute (NCI) NRSA Predoctoral Fellowship to Promote Diversity in Health-Related Research (F31CA228289). M.P.W. received support from the Lymphoma Research Foundation (337444) and the NIH (T32-CA009156-35). Y.N. was supported by grants-in-aid from the Japan Society for the Promotion of Science (JSPS) (15KK0356 and 16K11493). T.T. was supported by the Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study. M.V.G. was supported by Cancer Research UK (grants C7379/A15291 and C7379/A24639 to Mariann Bienz). The UNC Flow Cytometry Core Facility is supported in part by Cancer Center Core Support Grant P30 CA016086 to the UNC Lineberger Comprehensive Cancer Center, and research reported in this publication was supported by the Center for AIDS Research (award number 5P30AI050410), and the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, the Canada Foundation for Innovation, the Eshelman Institute for Innovation, Genome Canada, the Innovative Medicines Initiative (European Union [EU]/European Federation of Pharmaceutical Industries and Associations [EFPIA]) (ULTRA-DD grant no. 115766), Janssen, Merck & Company, Merck KGaA, Novartis Pharma AG, the Ontario Ministry of Economic Development and Innovation, Pfizer, the S{\~a}o Paulo Research Foundation (FAPESP) (2013/50724-5), Takeda, and the Wellcome Trust (106169/ZZ14/Z). R.R.R. received financial support from FAPESP (2016/17469-0). We would also like to thank Claire Strain-Damerell and Pavel Savitsky for cloning various mutants of AAK1 and BMP2K proteins that were used in the crystallization trials. Additionally, we thank Dr. Sean Conner for providing the AAK1 plasmids, Dr. Stephane Angers for kindly providing the HEK293T DVL TKO cells, and Dr. Mariann Bienz for providing comments and feedback. We would like to thank members of the Major laboratory for their feedback and expertise regarding experimental design and project direction. Publisher Copyright: {\textcopyright} 2018 The Authors",

year = "2019",

month = jan,

day = "2",

doi = "10.1016/j.celrep.2018.12.023",

language = "English",

volume = "26",

pages = "79--93.e8",

journal = "Cell Reports",

issn = "2211-1247",

number = "1",

}

Agajanian, MJ, Walker, MP, Axtman, AD, Ruela-de-Sousa, RR, Serafin, DS, Rabinowitz, AD, Graham, DM, Ryan, MB, Tamir, T, Nakamichi, Y, Gammons, MV, Bennett, JM, Couñago, RM, Drewry, DH, Elkins, JM, Gileadi, C, Gileadi, O, Godoi, PH, Kapadia, N, Müller, S, Santiago, AS, Sorrell, FJ, Wells, CI, Fedorov, O, Willson, TM, Zuercher, WJ & Major, MB 2019, 'WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop', Cell Reports, vol. 26, no. 1, pp. 79-93.e8. https://doi.org/10.1016/j.celrep.2018.12.023

TY - JOUR

T1 - WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop

AU - Agajanian, Megan J.

AU - Walker, Matthew P.

AU - Axtman, Alison D.

AU - Ruela-de-Sousa, Roberta R.

AU - Serafin, D. Stephen

AU - Rabinowitz, Alex D.

AU - Graham, David M.

AU - Ryan, Meagan B.

AU - Tamir, Tigist

AU - Nakamichi, Yuko

AU - Gammons, Melissa V.

AU - Bennett, James M.

AU - Couñago, Rafael M.

AU - Drewry, David H.

AU - Elkins, Jonathan M.

AU - Gileadi, Carina

AU - Gileadi, Opher

AU - Godoi, Paulo H.

AU - Kapadia, Nirav

AU - Müller, Susanne

AU - Santiago, André S.

AU - Sorrell, Fiona J.

AU - Wells, Carrow I.

AU - Fedorov, Oleg

AU - Willson, Timothy M.

AU - Zuercher, William J.

AU - Major, Michael B.

N1 - Funding Information: M.B.M. acknowledges support from the NIH ( RO1-CA187799 and U24-DK116204-01 ). M.J.A. received financial support from NIH T32 Predoctoral Training Grants in Pharmacology ( T32-GM007040-43 and T32-GM007040-42 ), an Initiative for Maximizing Student Diversity Grant ( R25-GM055336-16 ), and the NIH National Cancer Institute (NCI) NRSA Predoctoral Fellowship to Promote Diversity in Health-Related Research ( F31CA228289 ). M.P.W. received support from the Lymphoma Research Foundation ( 337444 ) and the NIH ( T32-CA009156-35 ). Y.N. was supported by grants-in-aid from the Japan Society for the Promotion of Science (JSPS) ( 15KK0356 and 16K11493 ). T.T. was supported by the Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study. M.V.G. was supported by Cancer Research UK (grants C7379/A15291 and C7379/A24639 to Mariann Bienz). The UNC Flow Cytometry Core Facility is supported in part by Cancer Center Core Support Grant P30 CA016086 to the UNC Lineberger Comprehensive Cancer Center, and research reported in this publication was supported by the Center for AIDS Research (award number 5P30AI050410 ), and the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie , Bayer Pharma AG , Boehringer Ingelheim , the Canada Foundation for Innovation , the Eshelman Institute for Innovation , Genome Canada , the Innovative Medicines Initiative ( European Union [EU]/European Federation of Pharmaceutical Industries and Associations [EFPIA]) (ULTRA-DD grant no. 115766), Janssen , Merck & Company , Merck KGaA , Novartis Pharma AG , the Ontario Ministry of Economic Development and Innovation , Pfizer , the São Paulo Research Foundation (FAPESP) ( 2013/50724-5 ), Takeda , and the Wellcome Trust ( 106169/ZZ14/Z ). R.R.R. received financial support from FAPESP ( 2016/17469-0 ). We would also like to thank Claire Strain-Damerell and Pavel Savitsky for cloning various mutants of AAK1 and BMP2K proteins that were used in the crystallization trials. Additionally, we thank Dr. Sean Conner for providing the AAK1 plasmids, Dr. Stephane Angers for kindly providing the HEK293T DVL TKO cells, and Dr. Mariann Bienz for providing comments and feedback. We would like to thank members of the Major laboratory for their feedback and expertise regarding experimental design and project direction. Funding Information: M.B.M. acknowledges support from the NIH (RO1-CA187799 and U24-DK116204-01). M.J.A. received financial support from NIH T32 Predoctoral Training Grants in Pharmacology (T32-GM007040-43 and T32-GM007040-42), an Initiative for Maximizing Student Diversity Grant (R25-GM055336-16), and the NIH National Cancer Institute (NCI) NRSA Predoctoral Fellowship to Promote Diversity in Health-Related Research (F31CA228289). M.P.W. received support from the Lymphoma Research Foundation (337444) and the NIH (T32-CA009156-35). Y.N. was supported by grants-in-aid from the Japan Society for the Promotion of Science (JSPS) (15KK0356 and 16K11493). T.T. was supported by the Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study. M.V.G. was supported by Cancer Research UK (grants C7379/A15291 and C7379/A24639 to Mariann Bienz). The UNC Flow Cytometry Core Facility is supported in part by Cancer Center Core Support Grant P30 CA016086 to the UNC Lineberger Comprehensive Cancer Center, and research reported in this publication was supported by the Center for AIDS Research (award number 5P30AI050410), and the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, the Canada Foundation for Innovation, the Eshelman Institute for Innovation, Genome Canada, the Innovative Medicines Initiative (European Union [EU]/European Federation of Pharmaceutical Industries and Associations [EFPIA]) (ULTRA-DD grant no. 115766), Janssen, Merck & Company, Merck KGaA, Novartis Pharma AG, the Ontario Ministry of Economic Development and Innovation, Pfizer, the S?o Paulo Research Foundation (FAPESP) (2013/50724-5), Takeda, and the Wellcome Trust (106169/ZZ14/Z). R.R.R. received financial support from FAPESP (2016/17469-0). We would also like to thank Claire Strain-Damerell and Pavel Savitsky for cloning various mutants of AAK1 and BMP2K proteins that were used in the crystallization trials. Additionally, we thank Dr. Sean Conner for providing the AAK1 plasmids, Dr. Stephane Angers for kindly providing the HEK293T DVL TKO cells, and Dr. Mariann Bienz for providing comments and feedback. We would like to thank members of the Major laboratory for their feedback and expertise regarding experimental design and project direction. Funding Information: M.B.M. acknowledges support from the NIH (RO1-CA187799 and U24-DK116204-01). M.J.A. received financial support from NIH T32 Predoctoral Training Grants in Pharmacology (T32-GM007040-43 and T32-GM007040-42), an Initiative for Maximizing Student Diversity Grant (R25-GM055336-16), and the NIH National Cancer Institute (NCI) NRSA Predoctoral Fellowship to Promote Diversity in Health-Related Research (F31CA228289). M.P.W. received support from the Lymphoma Research Foundation (337444) and the NIH (T32-CA009156-35). Y.N. was supported by grants-in-aid from the Japan Society for the Promotion of Science (JSPS) (15KK0356 and 16K11493). T.T. was supported by the Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study. M.V.G. was supported by Cancer Research UK (grants C7379/A15291 and C7379/A24639 to Mariann Bienz). The UNC Flow Cytometry Core Facility is supported in part by Cancer Center Core Support Grant P30 CA016086 to the UNC Lineberger Comprehensive Cancer Center, and research reported in this publication was supported by the Center for AIDS Research (award number 5P30AI050410), and the content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, the Canada Foundation for Innovation, the Eshelman Institute for Innovation, Genome Canada, the Innovative Medicines Initiative (European Union [EU]/European Federation of Pharmaceutical Industries and Associations [EFPIA]) (ULTRA-DD grant no. 115766), Janssen, Merck & Company, Merck KGaA, Novartis Pharma AG, the Ontario Ministry of Economic Development and Innovation, Pfizer, the São Paulo Research Foundation (FAPESP) (2013/50724-5), Takeda, and the Wellcome Trust (106169/ZZ14/Z). R.R.R. received financial support from FAPESP (2016/17469-0). We would also like to thank Claire Strain-Damerell and Pavel Savitsky for cloning various mutants of AAK1 and BMP2K proteins that were used in the crystallization trials. Additionally, we thank Dr. Sean Conner for providing the AAK1 plasmids, Dr. Stephane Angers for kindly providing the HEK293T DVL TKO cells, and Dr. Mariann Bienz for providing comments and feedback. We would like to thank members of the Major laboratory for their feedback and expertise regarding experimental design and project direction. Publisher Copyright: © 2018 The Authors

PY - 2019/1/2

Y1 - 2019/1/2

N2 - WNT signal transduction is essential for normal development and contributes to many human diseases. Agajanian et al. used a kinase gain-of-function screen to show that WNT activates the AAK1 kinase to promote clathrin-mediated endocytosis of the WNT receptor. This work identifies an AAK-driven negative feedback loop that downregulates WNT signaling.

AB - WNT signal transduction is essential for normal development and contributes to many human diseases. Agajanian et al. used a kinase gain-of-function screen to show that WNT activates the AAK1 kinase to promote clathrin-mediated endocytosis of the WNT receptor. This work identifies an AAK-driven negative feedback loop that downregulates WNT signaling.

KW - AAK1

KW - AP2M1

KW - LRP6

KW - WNT signaling

KW - clathrin

KW - endocytosis

KW - feedback loop

KW - gain-of-function screen

KW - kinase

UR - http://www.scopus.com/inward/record.url?scp=85059112535&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2018.12.023

DO - 10.1016/j.celrep.2018.12.023

M3 - Article

C2 - 30605688

AN - SCOPUS:85059112535

SN - 2211-1247

VL - 26

SP - 79-93.e8

JO - Cell Reports

JF - Cell Reports

IS - 1

ER -

WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop

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