Functional Integration of the Conserved Domains of Shoc2 Scaffold

Jeoung, Myoungkun; Abdelmoti, Lina; Jang, Eun Ryoung; Kooi, Craig W. Vander; Galperin, Emilia

doi:10.1371/journal.pone.0066067

Cited by 25 publications

(50 citation statements)

References 59 publications

(68 reference statements)

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“…Our earlier studies suggested that, in addition to tethering and enforcing physical proximity between Ras and RAF-1, the Shoc2 function extends to the coordination and integration of positive and negative ERK1/2 feedback loops (41). Thus, we set out to identify the proteins of the Shoc2 feedback loops and the mechanism that regulates Shoc2 function.…”

Section: Resultsmentioning

confidence: 99%

HUWE1 Is a Molecular Link Controlling RAF-1 Activity Supported by the Shoc2 Scaffold

Jang

Shi

Bryant

et al. 2014

Molecular and Cellular Biology

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Scaffold proteins play a critical role in controlling the activity of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway. Shoc2 is a leucine-rich repeat scaffold protein that acts as a positive modulator of ERK1/2 signaling. However, the precise mechanism by which Shoc2 modulates the activity of the ERK1/2 pathway is unclear. Here we report the identification of the E3 ubiquitin ligase HUWE1 as a binding partner and regulator of Shoc2 function. HUWE1 mediates ubiquitination and, consequently, the levels of Shoc2. Additionally, we show that both Shoc2 and HUWE1 are necessary to control the levels and ubiquitination of the Shoc2 signaling partner, RAF-1. Depletion of HUWE1 abolishes RAF-1 ubiquitination, with corresponding changes in ERK1/2 pathway activity occurring. Our results indicate that the HUWE1-mediated ubiquitination of Shoc2 is the switch that regulates the transition from an active to an inactive state of the RAF-1 kinase. Taken together, our results demonstrate that HUWE1 is a novel player involved in regulating ERK1/2 signal transmission through the Shoc2 scaffold complex.

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Section: Resultsmentioning

confidence: 99%

HUWE1 Is a Molecular Link Controlling RAF-1 Activity Supported by the Shoc2 Scaffold

Jang

Shi

Bryant

et al. 2014

Molecular and Cellular Biology

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“…S3A) (Jeoung et al, 2013). We found that a fragment of Shoc2 corresponding to leucine-rich repeats (LRRs) 20 to 21 (ΔN-19) was sufficient for PSMC5 binding (Fig.…”

Section: Psmc5 Binds To Leucine-rich Repeat 21 Of Shoc2mentioning

confidence: 90%

“…Shoc2 interacts with its signaling partners, Ras and RAF-1 through the unstructured N-terminal domain and the leucine-rich curvature of Shoc2 is left available to interact with additional components of the scaffolding module Jeoung et al, 2013). To further investigate the composition and dynamics of the Shoc2 scaffold complex, we performed a yeast two-hybrid screen in the human adult and fetal heart library using full-length Shoc2 as a bait as described previously .…”

Section: Interaction Of Shoc2 With Psmc5mentioning

confidence: 99%

“…All other counterparts in the Shoc2 complex, including PSMC5, were not affected or manipulated in any way offering a clear interpretation of how the loss of PSMC5 in the Shoc2 complex affects the cellular functions regulated by Shoc2-mediated ERK1/2 signals. The stability of the Shoc2-Δ21-C mutant and its ability to bind Ras and RAF-1 were assessed in our previous studies and found to be comparable to that of the full-length Shoc2 (Jeoung et al, 2013). In order to prevent clonal variations due to the different sites of viral genome incorporation, we utilized pooled populations of cells in the following experiments.…”

Section: Psmc5 Regulates Egfr/raf-1-induced Cell Growth and Motilitymentioning

confidence: 99%

“…Shoc2-tRFP and its mutants were described previously (Galperin et al, 2012;Jeoung et al, 2013). The plasmid carrying full-length PSMC5 was obtained from Open Biosystems (Thermo Scientific).…”

Section: Yeast Two-hybrid Screening Assaysmentioning

confidence: 99%

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Spatial control of Shoc2 scaffold-mediated ERK1/2 signaling requires remodeling activity of the ATPase PSMC5

Jang

Shi

et al. 2015

Journal of Cell Science

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The scaffold protein Shoc2 accelerates activity of the ERK1 and ERK2 (ERK1/2, also known as MAPK3 and MAPK1) pathway. Mutations in Shoc2 result in Noonan-like RASopathy, a developmental disorder with a wide spectrum of symptoms. The amplitude of the ERK1/2 signals transduced through the complex is fine-tuned by the HUWE1-mediated ubiquitylation of Shoc2 and its signaling partner RAF-1. Here, we provide a mechanistic basis of how ubiquitylation of Shoc2 and RAF-1 is controlled. We demonstrate that the newly identified binding partner of Shoc2, the (AAA+) ATPase PSMC5, triggers translocation of Shoc2 to endosomes. At the endosomes, PSMC5 displaces the E3 ligase HUWE1 from the scaffolding complex to attenuate ubiquitylation of Shoc2 and RAF-1. We show that a RASopathy mutation that changes the subcellular distribution of Shoc2 leads to alterations in Shoc2 ubiquitylation due to the loss of accessibility to PSMC5. In summary, our results demonstrate that PSMC5 is a new and important player involved in regulating ERK1/2 signal transmission through the remodeling of Shoc2 scaffold complex in a spatially-defined manner.

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A NovelSHOC2Variant in Rasopathy

et al. 2014

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Rasopathies are a group of genetic disorders caused by germline mutations in multiple genes of the Extracellular signal-Regulated Kinases 1 and 2 (ERK1/2) pathway. The only previously identified missense mutation in SHOC2, a scaffold protein of the ERK1/2 pathway, led to Noonan-like syndrome with loose anagen hair. Here we report a novel mutation in SHOC2(c.519G>A; p.M173I) that leads to a Rasopathy with clinical features partially overlapping those occurring in Noonan and Cardio-Facio-Cutaneous syndromes. Studies to clarify the significance of this SHOC2 variant revealed that the mutant protein has impaired capacity to interact with protein phosphatase 1c (PP1c), leading to insufficient activation of RAF-1 kinase. This SHOC2 variant thus is unable to fully rescue ERK1/2 activity in cells depleted of endogenous SHOC2. We conclude that SHOC2 mutations can cause a spectrum of Rasopathy phenotypes in heterozygous individuals. Importantly, our work suggests that individuals with mild Rasopathy symptoms may be under-diagnosed.

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Functional Integration of the Conserved Domains of Shoc2 Scaffold

Cited by 25 publications

References 59 publications

HUWE1 Is a Molecular Link Controlling RAF-1 Activity Supported by the Shoc2 Scaffold

HUWE1 Is a Molecular Link Controlling RAF-1 Activity Supported by the Shoc2 Scaffold

Spatial control of Shoc2 scaffold-mediated ERK1/2 signaling requires remodeling activity of the ATPase PSMC5

A NovelSHOC2Variant in Rasopathy

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