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PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72

Loss of function mutations in the PTEN-induced kinase 1 (PINK1) kinase are causal for autosomal recessive Parkinson's disease (PD) whilst gain of function mutations in the LRRK2 kinase cause autosomal dominant PD. PINK1 indirectly regulates the phosphorylation of a subset of Rab GTPases at a co...

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Autores principales: Vieweg, Sophie, Mulholland, Katie, Bräuning, Bastian, Kachariya, Nitin, Lai, Yu-Chiang, Toth, Rachel, Singh, Pawan Kishor, Volpi, Ilaria, Sattler, Michael, Groll, Michael, Itzen, Aymelt, Muqit, Miratul M. K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7219890/
https://www.ncbi.nlm.nih.gov/pubmed/32227113
http://dx.doi.org/10.1042/BCJ20190664
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author Vieweg, Sophie
Mulholland, Katie
Bräuning, Bastian
Kachariya, Nitin
Lai, Yu-Chiang
Toth, Rachel
Singh, Pawan Kishor
Volpi, Ilaria
Sattler, Michael
Groll, Michael
Itzen, Aymelt
Muqit, Miratul M. K.
author_facet Vieweg, Sophie
Mulholland, Katie
Bräuning, Bastian
Kachariya, Nitin
Lai, Yu-Chiang
Toth, Rachel
Singh, Pawan Kishor
Volpi, Ilaria
Sattler, Michael
Groll, Michael
Itzen, Aymelt
Muqit, Miratul M. K.
author_sort Vieweg, Sophie
collection PubMed
description Loss of function mutations in the PTEN-induced kinase 1 (PINK1) kinase are causal for autosomal recessive Parkinson's disease (PD) whilst gain of function mutations in the LRRK2 kinase cause autosomal dominant PD. PINK1 indirectly regulates the phosphorylation of a subset of Rab GTPases at a conserved Serine111 (Ser111) residue within the SF3 motif. Using genetic code expansion technologies, we have produced stoichiometric Ser111-phosphorylated Rab8A revealing impaired interactions with its cognate guanine nucleotide exchange factor and GTPase activating protein. In a screen for Rab8A kinases we identify TAK1 and MST3 kinases that can efficiently phosphorylate the Switch II residue Threonine72 (Thr72) in a similar manner as LRRK2 in vitro. Strikingly, we demonstrate that Ser111 phosphorylation negatively regulates the ability of LRRK2 but not MST3 or TAK1 to phosphorylate Thr72 of recombinant nucleotide-bound Rab8A in vitro and demonstrate an interplay of PINK1- and LRRK2-mediated phosphorylation of Rab8A in transfected HEK293 cells. Finally, we present the crystal structure of Ser111-phosphorylated Rab8A and nuclear magnetic resonance structure of Ser111-phosphorylated Rab1B. The structures reveal that the phosphorylated SF3 motif does not induce any major changes, but may interfere with effector-Switch II interactions through intramolecular H-bond formation and/or charge effects with Arg79. Overall, we demonstrate antagonistic regulation between PINK1-dependent Ser111 phosphorylation and LRRK2-mediated Thr72 phosphorylation of Rab8A indicating a potential cross-talk between PINK1-regulated mitochondrial homeostasis and LRRK2 signalling that requires further investigation in vivo.
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spelling pubmed-72198902020-05-29 PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72 Vieweg, Sophie Mulholland, Katie Bräuning, Bastian Kachariya, Nitin Lai, Yu-Chiang Toth, Rachel Singh, Pawan Kishor Volpi, Ilaria Sattler, Michael Groll, Michael Itzen, Aymelt Muqit, Miratul M. K. Biochem J Molecular Bases of Health & Disease Loss of function mutations in the PTEN-induced kinase 1 (PINK1) kinase are causal for autosomal recessive Parkinson's disease (PD) whilst gain of function mutations in the LRRK2 kinase cause autosomal dominant PD. PINK1 indirectly regulates the phosphorylation of a subset of Rab GTPases at a conserved Serine111 (Ser111) residue within the SF3 motif. Using genetic code expansion technologies, we have produced stoichiometric Ser111-phosphorylated Rab8A revealing impaired interactions with its cognate guanine nucleotide exchange factor and GTPase activating protein. In a screen for Rab8A kinases we identify TAK1 and MST3 kinases that can efficiently phosphorylate the Switch II residue Threonine72 (Thr72) in a similar manner as LRRK2 in vitro. Strikingly, we demonstrate that Ser111 phosphorylation negatively regulates the ability of LRRK2 but not MST3 or TAK1 to phosphorylate Thr72 of recombinant nucleotide-bound Rab8A in vitro and demonstrate an interplay of PINK1- and LRRK2-mediated phosphorylation of Rab8A in transfected HEK293 cells. Finally, we present the crystal structure of Ser111-phosphorylated Rab8A and nuclear magnetic resonance structure of Ser111-phosphorylated Rab1B. The structures reveal that the phosphorylated SF3 motif does not induce any major changes, but may interfere with effector-Switch II interactions through intramolecular H-bond formation and/or charge effects with Arg79. Overall, we demonstrate antagonistic regulation between PINK1-dependent Ser111 phosphorylation and LRRK2-mediated Thr72 phosphorylation of Rab8A indicating a potential cross-talk between PINK1-regulated mitochondrial homeostasis and LRRK2 signalling that requires further investigation in vivo. Portland Press Ltd. 2020-05-15 2020-05-11 /pmc/articles/PMC7219890/ /pubmed/32227113 http://dx.doi.org/10.1042/BCJ20190664 Text en © 2020 The Author(s) https://creativecommons.org/licenses/by/4.0/ This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Molecular Bases of Health & Disease
Vieweg, Sophie
Mulholland, Katie
Bräuning, Bastian
Kachariya, Nitin
Lai, Yu-Chiang
Toth, Rachel
Singh, Pawan Kishor
Volpi, Ilaria
Sattler, Michael
Groll, Michael
Itzen, Aymelt
Muqit, Miratul M. K.
PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72
title PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72
title_full PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72
title_fullStr PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72
title_full_unstemmed PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72
title_short PINK1-dependent phosphorylation of Serine111 within the SF3 motif of Rab GTPases impairs effector interactions and LRRK2-mediated phosphorylation at Threonine72
title_sort pink1-dependent phosphorylation of serine111 within the sf3 motif of rab gtpases impairs effector interactions and lrrk2-mediated phosphorylation at threonine72
topic Molecular Bases of Health & Disease
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7219890/
https://www.ncbi.nlm.nih.gov/pubmed/32227113
http://dx.doi.org/10.1042/BCJ20190664
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