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Article ; Online: The Highs and Lows of FBXW7: New Insights into Substrate Affinity in Disease and Development.

de la Cova, Claire C

2023 Volume 12, Issue 17

Abstract: FBXW7 is a critical regulator of cell cycle, cell signaling, and development. A highly conserved F-box protein and component of the SKP1-Cullin-F-box (SCF) complex, FBXW7 functions as a recognition subunit within a Cullin-RING E3 ubiquitin ligase ... ...

Abstract	FBXW7 is a critical regulator of cell cycle, cell signaling, and development. A highly conserved F-box protein and component of the SKP1-Cullin-F-box (SCF) complex, FBXW7 functions as a recognition subunit within a Cullin-RING E3 ubiquitin ligase responsible for ubiquitinating substrate proteins and targeting them for proteasome-mediated degradation. In human cells, FBXW7 promotes degradation of a large number of substrate proteins, including many that impact disease, such as NOTCH1, Cyclin E, MYC, and BRAF. A central focus for investigation has been to understand the molecular mechanisms that allow the exquisite substrate specificity exhibited by FBXW7. Recent work has produced a clearer understanding of how FBXW7 physically interacts with both high-affinity and low-affinity substrates. We review new findings that provide insights into the consequences of "hotspot" missense mutations of
MeSH term(s)	Humans ; Animals ; F-Box-WD Repeat-Containing Protein 7/genetics ; Caenorhabditis elegans ; Cullin Proteins ; F-Box Proteins/genetics ; Cell Cycle
Chemical Substances	F-Box-WD Repeat-Containing Protein 7 ; Cullin Proteins ; F-Box Proteins ; FBXW7 protein, human
Language	English
Publishing date	2023-08-24
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12172141
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Article ; Online: The E3/E4 ubiquitin ligase UFD-2 suppresses normal and oncogenic signaling mediated by a Raf ortholog in

Townley, Robert / Deniaud, Augustin / Stacy, Kennedy S / Rodriguez Torres, Claudia S / Cheraghi, Fatemeh / Wicker, Nicole B / de la Cova, Claire C

Science signaling

2023 Volume 16, Issue 800, Page(s) eabq4355

Abstract: Signaling by the kinase cascade composed of Raf, MEK, and ERK is critical for animal development and is often inappropriately activated in human malignancies. We sought to identify factors that control signaling mediated by ... ...

Abstract	Signaling by the kinase cascade composed of Raf, MEK, and ERK is critical for animal development and is often inappropriately activated in human malignancies. We sought to identify factors that control signaling mediated by the
MeSH term(s)	Animals ; Humans ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans Proteins/genetics ; Cell Cycle Proteins ; Mammals ; Mitogen-Activated Protein Kinase Kinases ; Signal Transduction ; Ubiquitin-Protein Ligases/genetics ; Ubiquitins
Chemical Substances	Caenorhabditis elegans Proteins ; Cell Cycle Proteins ; Mitogen-Activated Protein Kinase Kinases (EC 2.7.12.2) ; SEL-10 protein, C elegans ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; Ubiquitins ; ufd-2 protein, C elegans (EC 6.3.2.-) ; lin-45 protein, C elegans (EC 2.7.11.1)
Language	English
Publishing date	2023-08-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2417226-1
ISSN	1937-9145 ; 1945-0877
ISSN (online)	1937-9145
ISSN	1945-0877
DOI	10.1126/scisignal.abq4355
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Article ; Online: Negative feedback by conserved kinases patterns the degradation of

de la Cova, Claire C / Townley, Robert / Greenwald, Iva

Development (Cambridge, England)

2020 Volume 147, Issue 24

Abstract: Activation of a canonical EGFR-Ras-Raf-ERK cascade initiates patterning of multipotent vulval precursor cells (VPCs) ... ...

Abstract	Activation of a canonical EGFR-Ras-Raf-ERK cascade initiates patterning of multipotent vulval precursor cells (VPCs) of
MeSH term(s)	Animals ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans/growth & development ; Caenorhabditis elegans Proteins/genetics ; Cell Cycle Proteins/genetics ; Cell Differentiation/genetics ; Feedback, Physiological ; Female ; Gene Expression Regulation, Developmental/genetics ; Glycogen Synthase Kinase 3/genetics ; Phosphotransferases/genetics ; Proteolysis ; Signal Transduction/genetics ; Ubiquitin-Protein Ligases ; Vulva/growth & development ; Vulva/metabolism ; raf Kinases/genetics
Chemical Substances	Caenorhabditis elegans Proteins ; Cell Cycle Proteins ; SEL-10 protein, C elegans ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; Phosphotransferases (EC 2.7.-) ; lin-45 protein, C elegans (EC 2.7.11.1) ; raf Kinases (EC 2.7.11.1) ; Glycogen Synthase Kinase 3 (EC 2.7.11.26)
Language	English
Publishing date	2020-12-23
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.195941
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Article ; Online: Widely Used Mutants of

Kodra, Albana / de la Cova, Claire / Gerhold, Abigail R / Johnston, Laura A

G3 (Bethesda, Md.)

2020 Volume 10, Issue 12, Page(s) 4707–4712

Abstract: The process of apoptosis in epithelia involves activation of caspases, delamination of cells, and degradation of cellular components. Corpses and cellular debris are then rapidly cleared from the tissue by phagocytic blood cells. In studies of ... ...

Abstract	The process of apoptosis in epithelia involves activation of caspases, delamination of cells, and degradation of cellular components. Corpses and cellular debris are then rapidly cleared from the tissue by phagocytic blood cells. In studies of the
MeSH term(s)	Animals ; Drosophila/genetics ; Drosophila Proteins/genetics ; Drosophila melanogaster/genetics ; Mutation ; Receptors, Immunologic ; Tumor Necrosis Factor-alpha
Chemical Substances	Drosophila Proteins ; Receptors, Immunologic ; Tumor Necrosis Factor-alpha ; phagocytosis receptor
Language	English
Publishing date	2020-12-03
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2629978-1
ISSN	2160-1836 ; 2160-1836
ISSN (online)	2160-1836
ISSN	2160-1836
DOI	10.1534/g3.120.401800
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Article ; Online: A Real-Time Biosensor for ERK Activity Reveals Signaling Dynamics during C. elegans Cell Fate Specification.

de la Cova, Claire / Townley, Robert / Regot, Sergi / Greenwald, Iva

Developmental cell

2017 Volume 42, Issue 5, Page(s) 542–553.e4

Abstract: Kinase translocation reporters (KTRs) are genetically encoded fluorescent activity sensors that convert kinase activity into a nucleocytoplasmic shuttling equilibrium for visualizing single-cell signaling dynamics. Here, we adapt the first-generation KTR ...

Abstract	Kinase translocation reporters (KTRs) are genetically encoded fluorescent activity sensors that convert kinase activity into a nucleocytoplasmic shuttling equilibrium for visualizing single-cell signaling dynamics. Here, we adapt the first-generation KTR for extracellular signal-regulated kinase (ERK) to allow easy implementation in vivo. This sensor, "ERK-nKTR," allows quantitative and qualitative assessment of ERK activity by analysis of individual nuclei and faithfully reports ERK activity during development and neural function in diverse cell contexts in Caenorhabditis elegans. Analysis of ERK activity over time in the vulval precursor cells, a well-characterized paradigm of epidermal growth factor receptor (EGFR)-Ras-ERK signaling, has identified dynamic features not evident from analysis of developmental endpoints alone, including pulsatile frequency-modulated signaling associated with proximity to the EGF source. The toolkit described here will facilitate studies of ERK signaling in other C. elegans contexts, and the design features will enable implementation of this technology in other multicellular organisms.
MeSH term(s)	Animals ; Biosensing Techniques/methods ; Caenorhabditis elegans/cytology ; Caenorhabditis elegans/enzymology ; Cell Line ; Cell Lineage ; Cell Movement ; Cell Nucleus/metabolism ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Female ; Genes, Reporter ; Germ Cells/cytology ; MAP Kinase Signaling System ; Mammals ; Mutation/genetics ; Myoblasts/cytology ; Neurons/cytology ; Phosphorylation ; Reproducibility of Results ; Stem Cells/cytology ; Stem Cells/metabolism ; Subcellular Fractions/metabolism ; Vulva/cytology
Chemical Substances	Extracellular Signal-Regulated MAP Kinases (EC 2.7.11.24)
Language	English
Publishing date	2017-08-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2054967-2
ISSN	1878-1551 ; 1534-5807
ISSN (online)	1878-1551
ISSN	1534-5807
DOI	10.1016/j.devcel.2017.07.014
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Article ; Online: SEL-10/Fbw7-dependent negative feedback regulation of LIN-45/Braf signaling in C. elegans via a conserved phosphodegron.

de la Cova, Claire / Greenwald, Iva

Genes & development

2012 Volume 26, Issue 22, Page(s) 2524–2535

Abstract: The conserved E3 ubiquitin ligase component named SEL-10 in Caenorhabditis elegans and Fbw7 in mammals targets substrates for ubiquitin-mediated degradation through a high-affinity binding site called a Cdc4 phosphodegron (CPD). As many known substrates ... ...

Abstract	The conserved E3 ubiquitin ligase component named SEL-10 in Caenorhabditis elegans and Fbw7 in mammals targets substrates for ubiquitin-mediated degradation through a high-affinity binding site called a Cdc4 phosphodegron (CPD). As many known substrates of Fbw7 are oncoproteins, the identification of new substrates may offer insight into cancer biology as well as aspects of proteome regulation. Here, we evaluated whether the presence of an evolutionarily conserved CPD would be a feasible complement to proteomics-based approaches for identifying new potential substrates. For functional assessments, we focused on LIN-45, a component of the signal transduction pathway underlying vulval induction and the ortholog of human Braf, an effector of Ras in numerous cancers. Our analysis demonstrates that LIN-45 behaves as a bona fide substrate of SEL-10, with mutation of the CPD or loss of sel-10 resulting in increased activity and protein stability in vivo. Furthermore, during vulval induction, the downstream kinase MPK-1/ERK is also required for LIN-45 protein degradation in a negative feedback loop, resulting in degradation of LIN-45 where ERK is highly active. As the CPD consensus sequence is conserved in human Braf, we propose that Fbw7 may also regulate Braf stability in some cell contexts. We discuss the implications of our findings for vulval development in C. elegans, the potential applicability to human Braf, and the value of a CPD-based predictive approach for human Fbw7 substrates.
MeSH term(s)	Animals ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans/metabolism ; Caenorhabditis elegans/physiology ; Caenorhabditis elegans Proteins/genetics ; Caenorhabditis elegans Proteins/metabolism ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Down-Regulation ; Feedback, Physiological ; Humans ; Protein Binding ; Proto-Oncogene Proteins B-raf/genetics ; Proto-Oncogene Proteins B-raf/metabolism ; Signal Transduction/genetics ; Ubiquitin-Protein Ligases/genetics ; Ubiquitin-Protein Ligases/metabolism ; Vertebrates/genetics ; raf Kinases/genetics ; raf Kinases/metabolism
Chemical Substances	Caenorhabditis elegans Proteins ; Cell Cycle Proteins ; SEL-10 protein, C elegans ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; Proto-Oncogene Proteins B-raf (EC 2.7.11.1) ; lin-45 protein, C elegans (EC 2.7.11.1) ; raf Kinases (EC 2.7.11.1)
Language	English
Publishing date	2012-11-15
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	806684-x
ISSN	1549-5477 ; 0890-9369
ISSN (online)	1549-5477
ISSN	0890-9369
DOI	10.1101/gad.203703.112
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Article: Myc in model organisms: a view from the flyroom.

de la Cova, Claire / Johnston, Laura A

Seminars in cancer biology

2006 Volume 16, Issue 4, Page(s) 303–312

Abstract: The Myc transcription factor regulates fundamental processes in a cell's life: its growth, division, and survival. Myc is conserved throughout metazoan phyla, and its identification in the fruit fly, Drosophila melanogaster has led to new insights in Myc' ...

Abstract	The Myc transcription factor regulates fundamental processes in a cell's life: its growth, division, and survival. Myc is conserved throughout metazoan phyla, and its identification in the fruit fly, Drosophila melanogaster has led to new insights in Myc's physiological roles. In this review, we describe recent research on the biology of Myc and its family members in Drosophila, paying particular attention to its role in the control of growth during development.
MeSH term(s)	Animals ; Animals, Genetically Modified ; Drosophila Proteins/physiology ; Drosophila melanogaster/physiology ; Gene Expression Regulation, Developmental ; Proto-Oncogene Proteins c-myc/physiology ; Signal Transduction ; Trans-Activators
Chemical Substances	Drosophila Proteins ; Proto-Oncogene Proteins c-myc ; Trans-Activators
Language	English
Publishing date	2006-08
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1033980-2
ISSN	1096-3650 ; 1044-579X
ISSN (online)	1096-3650
ISSN	1044-579X
DOI	10.1016/j.semcancer.2006.07.010
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Article ; Online: Supercompetitor status of Drosophila Myc cells requires p53 as a fitness sensor to reprogram metabolism and promote viability.

de la Cova, Claire / Senoo-Matsuda, Nanami / Ziosi, Marcello / Wu, D Christine / Bellosta, Paola / Quinzii, Catarina M / Johnston, Laura A

Cell metabolism

2014 Volume 19, Issue 3, Page(s) 470–483

Abstract: In growing tissues, cell fitness disparities can provoke interactions that promote stronger cells at the expense of the weaker in a process called cell competition. The mechanistic definition of cell fitness is not understood, nor is it understood how ... ...

Abstract	In growing tissues, cell fitness disparities can provoke interactions that promote stronger cells at the expense of the weaker in a process called cell competition. The mechanistic definition of cell fitness is not understood, nor is it understood how fitness differences are recognized. Drosophila cells with extra Myc activity acquire "supercompetitor" status upon confrontation with wild-type (WT) cells, prompting the latter's elimination via apoptosis. Here we show that such confrontation enhances glycolytic flux in Myc cells and promotes their fitness and proliferation in a p53-dependent manner. Whereas p53 loss in noncompeting Myc cells is inconsequential, its loss impairs metabolism, reduces viability, and prevents the killing activity of Myc supercompetitor cells. We propose that p53 acts as a general sensor of competitive confrontation to enhance the fitness of the "winner" population. Our findings suggest that the initial confrontation between precancerous and WT cells could enhance cancer cell fitness and promote tumor progression.
MeSH term(s)	Animals ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Drosophila/cytology ; Drosophila/metabolism ; Drosophila Proteins/genetics ; Drosophila Proteins/metabolism ; Genomic Instability ; Glycolysis ; Oxidative Phosphorylation ; Proto-Oncogene Proteins c-myc/genetics ; Proto-Oncogene Proteins c-myc/metabolism ; RNA, Messenger/metabolism ; Tumor Suppressor Protein p53/genetics ; Tumor Suppressor Protein p53/metabolism
Chemical Substances	Drosophila Proteins ; Proto-Oncogene Proteins c-myc ; RNA, Messenger ; Tumor Suppressor Protein p53
Language	English
Publishing date	2014-02-20
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2176834-1
ISSN	1932-7420 ; 1550-4131
ISSN (online)	1932-7420
ISSN	1550-4131
DOI	10.1016/j.cmet.2014.01.012
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Article: Drosophila myc regulates organ size by inducing cell competition.

de la Cova, Claire / Abril, Mauricio / Bellosta, Paola / Gallant, Peter / Johnston, Laura A

Cell

2004 Volume 117, Issue 1, Page(s) 107–116

Abstract: Experiments in both vertebrates and invertebrates have illustrated the competitive nature of growth and led to the idea that competition is a mechanism of regulating organ and tissue size. We have assessed competitive interactions between cells in a ... ...

Abstract	Experiments in both vertebrates and invertebrates have illustrated the competitive nature of growth and led to the idea that competition is a mechanism of regulating organ and tissue size. We have assessed competitive interactions between cells in a developing organ and examined their effect on its final size. We show that local expression of the Drosophila growth regulator dMyc, a homolog of the c-myc protooncogene, induces cell competition and leads to the death of nearby wild-type cells in developing wings. We demonstrate that cell competition is executed via induction of the proapoptotic gene hid and that both competition and hid function are required for the wing to reach an appropriate size when dMyc is expressed. Moreover, we provide evidence that reproducible wing size during normal development requires apoptosis. Modulating dmyc levels to create cell competition and hid-dependent cell death may be a mechanism used during normal development to control organ size.
MeSH term(s)	Animals ; Apoptosis/physiology ; Body Patterning/genetics ; Body Weight/genetics ; Cell Communication/genetics ; Cell Differentiation/genetics ; Cells, Cultured ; Clone Cells ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Drosophila Proteins/genetics ; Drosophila Proteins/metabolism ; Drosophila melanogaster/genetics ; Drosophila melanogaster/growth & development ; Drosophila melanogaster/metabolism ; Gene Expression Regulation, Developmental/genetics ; Morphogenesis/genetics ; Neuropeptides/genetics ; Neuropeptides/metabolism ; Organogenesis/genetics ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Wings, Animal/cytology ; Wings, Animal/growth & development ; Wings, Animal/metabolism
Chemical Substances	DNA-Binding Proteins ; Drosophila Proteins ; HID protein, Drosophila ; Myc protein, Drosophila ; Neuropeptides ; Transcription Factors
Language	English
Publishing date	2004-04-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	187009-9
ISSN	1097-4172 ; 0092-8674
ISSN (online)	1097-4172
ISSN	0092-8674
DOI	10.1016/s0092-8674(04)00214-4
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Article: Analysis of free hydroxytyrosol in human plasma following the administration of olive oil

Pastor, Antoni / Jose RodrÃguez-MoratÃ³ / EulÃ lia Olesti / Mitona Pujadas / Clara PÃ©rez-MaÃ±Ã¡ / Olha Khymenets / Montserrat FitÃ³ / MarÃa-Isabel Covas / Rosa SolÃ¡ / MarÃa-JosÃ© Motilva / MagÃ FarrÃ© / Rafael de la Torre

Journal of chromatography. 2016 Mar. 11, v. 1437

2016

Abstract: Hydroxytyrosol (HT) from olive oil, a potent bioactive molecule with health benefits, has a poor bioavailability, its free form (free HT) being undetectable so far. This fact leads to the controversy whether attained HT concentrations after olive oil ... ...

Abstract	Hydroxytyrosol (HT) from olive oil, a potent bioactive molecule with health benefits, has a poor bioavailability, its free form (free HT) being undetectable so far. This fact leads to the controversy whether attained HT concentrations after olive oil polyphenol ingestion are too low to explain the observed biological activities. Due to this, an analytical methodology to determine free HT in plasma is crucial for understanding HT biological activity. Plasma HT instability and low concentrations have been major limitations for its quantification in clinical studies. Here, we describe a method to detect and quantify free HT in human plasma by using liquid chromatography coupled to tandem mass spectrometry. The method encompasses different steps of sample preparation including plasma stabilization, protein precipitation, selective derivatization with benzylamine, and purification by solid-phase extraction. A high sensitivity (LOD, 0.3ng/mL), specificity and stability of HT is achieved following these procedures. The method was validated and its applicability was demonstrated by analyzing human plasma samples after olive oil intake. A pharmacokinetic comparison was performed measuring free HT plasma concentrations following the intake of 25mL of ordinary olive oil (nearly undetectable concentrations) versus an extra-virgin olive oil (Cmax=4.40ng/mL). To our knowledge, this is the first time that an analytical procedure for quantifying free HT in plasma after olive oil dietary doses has been reported. The present methodology opens the door to a better understanding of the relationship between HT plasma concentrations and its beneficial health effects.
Keywords	bioactive properties ; bioavailability ; clinical trials ; derivatization ; extra-virgin olive oil ; humans ; ingestion ; liquid chromatography ; pharmacokinetics ; polyphenols ; solid phase extraction ; tandem mass spectrometry
Language	English
Dates of publication	2016-0311
Size	p. 183-190.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	218139-3
ISSN	0021-9673 ; 0378-4355 ; 0376-737X
ISSN	0021-9673 ; 0378-4355 ; 0376-737X
DOI	10.1016/j.chroma.2016.02.016
Database	NAL-Catalogue (AGRICOLA)

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