Mapping the binding interface of ERK and transcriptional repressor Capicua using photocrosslinking

Futran, Alan; Kyin, Saw; Shvartsman, Stanislav Y.; Link, A. James

doi:10.1073/pnas.1501373112

Cited by 49 publications

(56 citation statements)

References 50 publications

(63 reference statements)

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“…We found that the subcellular distribution of Cic and expression of its target genes were unaffected in CSN1b or CSN5 mutant follicle cells (data not shown), suggesting that Cic is not ubiquitinated, and therefore does not require CSN function, in this tissue. Conversely, our data show that Cic stability can be regulated independently of MAPK and of the C2 domain that is required for MAPK binding (Astigarraga et al, 2007;Futran et al, 2015), through a mechanism that is specifically antagonized by CSN1b. The competition between CSN1b and this alternative mode of degradation would establish the basal level of Cic in the absence of RTK activity.…”

Section: Regulation Of Capicua Degradationcontrasting

confidence: 51%

“…Activated MAPK phosphorylates Cic after binding to its conserved C2 domain; phosphorylation of potential target sites in human CIC interferes with binding of its nuclear localization signal to importin-α (Astigarraga et al, 2007;Dissanayake et al, 2011;Futran et al, 2015). Consistent with this model, EGFR signaling results in cytoplasmic localization of Cic (Astigarraga et al, 2007), accompanied in some tissues by its degradation (Roch et al, 2002).…”

Section: Introductionmentioning

confidence: 94%

“…In the wing disc, MAPK promotes aos expression by binding to and phosphorylating the transcriptional repressor Cic, inhibiting its function and triggering its degradation ( Fig. 3A) (Astigarraga et al, 2007;Futran et al, 2015;Roch et al, 2002). This makes Cic a good candidate to mediate the effect of the CSN on EGFR signaling.…”

Section: The Cop9 Signalosome Promotes Capicua Accumulationmentioning

confidence: 99%

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COP9 signalosome subunits protect Capicua from MAPK-dependent and -independent mechanisms of degradation

Suisse¹,

He²,

Legent³

et al. 2017

Development

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The COP9 signalosome removes Nedd8 modifications from the Cullin subunits of ubiquitin ligase complexes, reducing their activity. Here, we show that mutations in the Drosophila COP9 signalosome subunit 1b (CSN1b) gene increase the activity of ubiquitin ligases that contain Cullin 1. Analysis of CSN1b mutant phenotypes revealed a requirement for the COP9 signalosome to prevent ectopic expression of Epidermal growth factor receptor (EGFR) target genes. It does so by protecting Capicua, a transcriptional repressor of EGFR target genes, from EGFR pathway-dependent ubiquitylation by a Cullin 1/SKP1-related A/Archipelago E3 ligase and subsequent proteasomal degradation. The CSN1b subunit also maintains basal Capicua levels by protecting it from a separate mechanism of degradation that is independent of EGFR signaling. As a suppressor of tumor growth and metastasis, Capicua may be an important target of the COP9 signalosome in cancer.

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Section: Regulation Of Capicua Degradationcontrasting

confidence: 51%

Section: Introductionmentioning

confidence: 94%

Section: The Cop9 Signalosome Promotes Capicua Accumulationmentioning

confidence: 99%

See 1 more Smart Citation

COP9 signalosome subunits protect Capicua from MAPK-dependent and -independent mechanisms of degradation

Suisse¹,

He²,

Legent³

et al. 2017

Development

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“…ERK-mediated down-regulation of Cic depends on the conserved C2 motif located in region Cic3, which serves as the ERK docking site (5). Deletion of the C2 motif abrogated Cic-ERK interaction (5), and a single amino acid substitution, F1054A, in the C2 motif (QQFILAPTPAQLG) reduced the binding of Cic to ERK (26). Importantly, deletion of the C2 domain did not affect the binding of Cic to Mnb (Fig.…”

Section: Significancementioning

confidence: 91%

“…2C). Our previous studies showed that region Cic3 includes an ERK docking site and is subject to ERK-mediated phosphorylation (5,14,26). To compare the activities of Mnb and ERK, we coexpressed Cic1 or Cic3 with Mnb or a constitutively active Drosophila ERK, ERK Sem (27).…”

Section: Significancementioning

confidence: 99%

Minibrain and Wings apart control organ growth and tissue patterning through down-regulation of Capicua

Liu

Paul

Trieu

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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The transcriptional repressor Capicua (Cic) controls tissue patterning and restricts organ growth, and has been recently implicated in several cancers. Cic has emerged as a primary sensor of signaling downstream of the receptor tyrosine kinase (RTK)/extracellular signal-regulated kinase (ERK) pathway, but how Cic activity is regulated in different cellular contexts remains poorly understood. We found that the kinase Minibrain (Mnb, ortholog of mammalian DYRK1A), acting through the adaptor protein Wings apart (Wap), physically interacts with and phosphorylates the Cic protein. Mnb and Wap inhibit Cic function by limiting its transcriptional repressor activity. Down-regulation of Cic by Mnb/Wap is necessary for promoting the growth of multiple organs, including the wings, eyes, and the brain, and for proper tissue patterning in the wing. We have thus uncovered a previously unknown mechanism of down-regulation of Cic activity by Mnb and Wap, which operates independently from the ERK-mediated control of Cic. Therefore, Cic functions as an integrator of upstream signals that are essential for tissue patterning and organ growth. Finally, because DYRK1A and CIC exhibit, respectively, prooncogenic vs. tumor suppressor activities in human oligodendroglioma, our results raise the possibility that DYRK1A may also down-regulate CIC in human cells.

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Assessing the predictive capabilities of design heuristics and coarse‐grain simulation toward understanding and optimizing site‐specific covalent immobilization of β‐lactamase

Soltani

Hunt

Smith

et al. 2022

Biotechnology Journal

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For industrial applications, covalent immobilization of enzymes provides minimum leakage, recoverability, reusability, and high stability. Yet, the suitability of a given site on the enzyme for immobilization remains a trial‐and‐error procedure. Here, we investigate the reliability of design heuristics and a coarse‐grain molecular simulation in predicting the optimum sites for covalent immobilization of TEM‐1 β‐lactamase. We utilized Escherichia coli‐lysate‐based cell‐free protein synthesis (CFPS) to produce variants containing a site‐specific incorporated unnatural amino acid with a unique moiety to facilitate site directed covalent immobilization. To constrain the number of potential immobilization sites, we investigated the predictive capability of several design heuristics. The suitability of immobilization sites was determined by analyzing expression yields, specific activity, immobilization efficiency, and stability of variants. These experimental findings are compared with coarse‐grain simulation of TEM‐1 domain stability and thermal stability and analyzed for a priori predictive capabilities. This work demonstrates that the design heuristics successfully identify a subset of locations for experimental validation. Specifically, the nucleotide following amber stop codon and domain stability correlate well with the expression yield and specific activity of the variants, respectively. Our approach highlights the advantages of combining coarse‐grain simulation and high‐throughput experimentation using CFPS to identify optimal enzyme immobilization sites.

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Mapping the binding interface of ERK and transcriptional repressor Capicua using photocrosslinking

Cited by 49 publications

References 50 publications

COP9 signalosome subunits protect Capicua from MAPK-dependent and -independent mechanisms of degradation

COP9 signalosome subunits protect Capicua from MAPK-dependent and -independent mechanisms of degradation

Minibrain and Wings apart control organ growth and tissue patterning through down-regulation of Capicua

Assessing the predictive capabilities of design heuristics and coarse‐grain simulation toward understanding and optimizing site‐specific covalent immobilization of β‐lactamase

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