SHOC2 subcellular shuttling requires the KEKE motif-rich region and N-terminal leucine-rich repeat domain and impacts on ERK signalling

Marialetizia Motta, Giovanni Chillemi, Valentina Fodale, Serena Cecchetti, Simona Coppola, Silvia Stipo, Viviana Cordeddu, Pompeo Macioce, Bruce D. Gelb, Marco Tartaglia

Research output: Contribution to journalArticle

Abstract

SHOC2 is a scaffold protein composed almost entirely by leucine-rich repeats (LRRs) and having an N-terminal region enriched in alternating lysine and glutamate/aspartate residues (KEKE motifs). SHOC2 acts as a positive modulator of the RAS-RAF-MEK-ERK signalling cascade by favouring stable RAF1 interaction with RAS. We previously reported that the p.Ser2Gly substitution in SHOC2 underlies Mazzanti syndrome, a RASopathy clinically overlapping Noonan syndrome, promoting N-myristoylation and constitutive targeting of the mutant to the plasma membrane. We also documented transient nuclear translocation of wild-type SHOC2 upon EGF stimulation, suggesting a more complex function in signal transduction.Here, we characterized the domains controlling SHOC2 shuttling between the nucleus and cytoplasm, and those contributing to SHOC2(S2G) mistargeting to the plasma membrane, analysed the structural organization of SHOC2's LRR motifs, and determined the impact of SHOC2 mislocalization on ERK signalling. We show that LRRs 1 to 13 constitute a structurally recognizable domain required for SHOC2 import into the nucleus and constitutive targeting of SHOC2(S2G) to the plasma membrane, while the KEKE motif-rich region is necessary to achieve efficient SHOC2 export from the nucleus. We also document that SHOC2(S2G) localizes both in raft and non-raft domains, and that it translocates to the non-raft domains following stimulation. Finally, we demonstrate that SHOC2 trapping at different subcellular sites has a diverse impact on ERK signalling strength and dynamics, suggesting a dual counteracting modulatory role of SHOC2 in the control of ERK signalling exerted at different intracellular compartments.

Original languageEnglish
Pages (from-to)3824-3835
Number of pages12
JournalHuman Molecular Genetics
Volume25
Issue number17
DOIs
Publication statusPublished - Sep 1 2016

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Leucine
Cell Membrane
Noonan Syndrome
Mitogen-Activated Protein Kinase Kinases
Epidermal Growth Factor
Aspartic Acid
Lysine
Glutamic Acid
Signal Transduction
Cytoplasm
Proteins

Keywords

  • Journal Article

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SHOC2 subcellular shuttling requires the KEKE motif-rich region and N-terminal leucine-rich repeat domain and impacts on ERK signalling. / Motta, Marialetizia; Chillemi, Giovanni; Fodale, Valentina; Cecchetti, Serena; Coppola, Simona; Stipo, Silvia; Cordeddu, Viviana; Macioce, Pompeo; Gelb, Bruce D.; Tartaglia, Marco.

In: Human Molecular Genetics, Vol. 25, No. 17, 01.09.2016, p. 3824-3835.

Research output: Contribution to journalArticle

Motta, M, Chillemi, G, Fodale, V, Cecchetti, S, Coppola, S, Stipo, S, Cordeddu, V, Macioce, P, Gelb, BD & Tartaglia, M 2016, 'SHOC2 subcellular shuttling requires the KEKE motif-rich region and N-terminal leucine-rich repeat domain and impacts on ERK signalling', Human Molecular Genetics, vol. 25, no. 17, pp. 3824-3835. https://doi.org/10.1093/hmg/ddw229
Motta, Marialetizia ; Chillemi, Giovanni ; Fodale, Valentina ; Cecchetti, Serena ; Coppola, Simona ; Stipo, Silvia ; Cordeddu, Viviana ; Macioce, Pompeo ; Gelb, Bruce D. ; Tartaglia, Marco. / SHOC2 subcellular shuttling requires the KEKE motif-rich region and N-terminal leucine-rich repeat domain and impacts on ERK signalling. In: Human Molecular Genetics. 2016 ; Vol. 25, No. 17. pp. 3824-3835.
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T1 - SHOC2 subcellular shuttling requires the KEKE motif-rich region and N-terminal leucine-rich repeat domain and impacts on ERK signalling

AU - Motta, Marialetizia

AU - Chillemi, Giovanni

AU - Fodale, Valentina

AU - Cecchetti, Serena

AU - Coppola, Simona

AU - Stipo, Silvia

AU - Cordeddu, Viviana

AU - Macioce, Pompeo

AU - Gelb, Bruce D.

AU - Tartaglia, Marco

N1 - © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

PY - 2016/9/1

Y1 - 2016/9/1

N2 - SHOC2 is a scaffold protein composed almost entirely by leucine-rich repeats (LRRs) and having an N-terminal region enriched in alternating lysine and glutamate/aspartate residues (KEKE motifs). SHOC2 acts as a positive modulator of the RAS-RAF-MEK-ERK signalling cascade by favouring stable RAF1 interaction with RAS. We previously reported that the p.Ser2Gly substitution in SHOC2 underlies Mazzanti syndrome, a RASopathy clinically overlapping Noonan syndrome, promoting N-myristoylation and constitutive targeting of the mutant to the plasma membrane. We also documented transient nuclear translocation of wild-type SHOC2 upon EGF stimulation, suggesting a more complex function in signal transduction.Here, we characterized the domains controlling SHOC2 shuttling between the nucleus and cytoplasm, and those contributing to SHOC2(S2G) mistargeting to the plasma membrane, analysed the structural organization of SHOC2's LRR motifs, and determined the impact of SHOC2 mislocalization on ERK signalling. We show that LRRs 1 to 13 constitute a structurally recognizable domain required for SHOC2 import into the nucleus and constitutive targeting of SHOC2(S2G) to the plasma membrane, while the KEKE motif-rich region is necessary to achieve efficient SHOC2 export from the nucleus. We also document that SHOC2(S2G) localizes both in raft and non-raft domains, and that it translocates to the non-raft domains following stimulation. Finally, we demonstrate that SHOC2 trapping at different subcellular sites has a diverse impact on ERK signalling strength and dynamics, suggesting a dual counteracting modulatory role of SHOC2 in the control of ERK signalling exerted at different intracellular compartments.

AB - SHOC2 is a scaffold protein composed almost entirely by leucine-rich repeats (LRRs) and having an N-terminal region enriched in alternating lysine and glutamate/aspartate residues (KEKE motifs). SHOC2 acts as a positive modulator of the RAS-RAF-MEK-ERK signalling cascade by favouring stable RAF1 interaction with RAS. We previously reported that the p.Ser2Gly substitution in SHOC2 underlies Mazzanti syndrome, a RASopathy clinically overlapping Noonan syndrome, promoting N-myristoylation and constitutive targeting of the mutant to the plasma membrane. We also documented transient nuclear translocation of wild-type SHOC2 upon EGF stimulation, suggesting a more complex function in signal transduction.Here, we characterized the domains controlling SHOC2 shuttling between the nucleus and cytoplasm, and those contributing to SHOC2(S2G) mistargeting to the plasma membrane, analysed the structural organization of SHOC2's LRR motifs, and determined the impact of SHOC2 mislocalization on ERK signalling. We show that LRRs 1 to 13 constitute a structurally recognizable domain required for SHOC2 import into the nucleus and constitutive targeting of SHOC2(S2G) to the plasma membrane, while the KEKE motif-rich region is necessary to achieve efficient SHOC2 export from the nucleus. We also document that SHOC2(S2G) localizes both in raft and non-raft domains, and that it translocates to the non-raft domains following stimulation. Finally, we demonstrate that SHOC2 trapping at different subcellular sites has a diverse impact on ERK signalling strength and dynamics, suggesting a dual counteracting modulatory role of SHOC2 in the control of ERK signalling exerted at different intracellular compartments.

KW - Journal Article

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DO - 10.1093/hmg/ddw229

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VL - 25

SP - 3824

EP - 3835

JO - Human Molecular Genetics

JF - Human Molecular Genetics

SN - 0964-6906

IS - 17

ER -