Structure of phospholipase Cε reveals an integrated RA1 domain and previously unidentified regulatory elements

Rugema, Ngango Yvon; Garland‐Kuntz, Elisabeth E.; Sieng, Monita; Muralidharan, Kaushik; Camp, Michelle M. Van; O'Neill, Hannah; Mbongo, William; Selvia, Arielle F.; Marti, Andrea; Everly, Amanda; McKenzie, Emmanda; Lyon, Angeline M.

doi:10.1038/s42003-020-01178-8

Cited by 13 publications

(12 citation statements)

References 55 publications

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“…While this phospholipase contains a C2 domain that could direct the protein to the PM, a recent 3D structure of (almost) full-length PLCE1 provided insights into its mechanisms and PM localization [54]. The model proposed that the C2 domain was not used for PM binding, but rather the αX-Y helix in the lipase domain.…”

Section: Plce1mentioning

confidence: 99%

The Ins and Outs of RAS Effector Complexes

2021

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RAS oncogenes are among the most commonly mutated proteins in human cancers. They regulate a wide range of effector pathways that control cell proliferation, survival, differentiation, migration and metabolic status. Including aberrations in these pathways, RAS-dependent signaling is altered in more than half of human cancers. Targeting mutant RAS proteins and their downstream oncogenic signaling pathways has been elusive. However, recent results comprising detailed molecular studies, large scale omics studies and computational modeling have painted a new and more comprehensive portrait of RAS signaling that helps us to understand the intricacies of RAS, how its physiological and pathophysiological functions are regulated, and how we can target them. Here, we review these efforts particularly trying to relate the detailed mechanistic studies with global functional studies. We highlight the importance of computational modeling and data integration to derive an actionable understanding of RAS signaling that will allow us to design new mechanism-based therapies for RAS mutated cancers.

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Section: Plce1mentioning

confidence: 99%

The Ins and Outs of RAS Effector Complexes

2021

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“…The PH-COOH Y2155A, L2158A, L2192A, and F2198A mutants all had T m values comparable with that of PH-COOH ( 15 , 30 ), and basal specific activities within ∼2-fold of PH-COOH ( Table 1 and Fig. S1 ), demonstrating that they are properly folded.…”

Section: Resultsmentioning

confidence: 85%

“…The PLCε RA domains are highly similar in structure but have different functional roles in the enzyme ( 1 , 6 , 15 , 16 , 17 , 18 ). The RA1 domain, together with the C2-RA1 linker, forms extensive contacts with EF3/4, the TIM barrel, and the C2 domain that are important for stability and activity ( 15 ). RA1 also interacts with mAKAP, a scaffolding protein at the perinuclear membrane, helping localize PLCε to internal membranes ( 18 ).…”

Section: Discussionmentioning

confidence: 99%

“…RA1 also interacts with mAKAP, a scaffolding protein at the perinuclear membrane, helping localize PLCε to internal membranes ( 18 ). The contribution of the RA2 domain to basal activity is unclear, because its deletion has been reported to either activate, inhibit, or have minimal impact ( 14 , 15 , 17 ). The RA2 domain is the primary binding site for activated Rap1A and Ras GTPases, because deletion of the domain or mutation of two highly conserved lysines ( R. norvegicus PLCε Lys 2150 and Lys 2152 ; Fig.…”

Section: Discussionmentioning

confidence: 99%

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Functional and structural characterization of allosteric activation of phospholipase Cε by Rap1A

Sieng

Selvia

Garland‐Kuntz

et al. 2020

Journal of Biological Chemistry

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Phospholipase Cε (PLCe) is activated downstream of G protein-coupled receptors (GPCRs) and receptor tyrosine kinases (RTKs) through direct interactions with small GTPases, including Rap1A and Ras. While Ras has been reported to allosterically activate the lipase, it is not known whether Rap1A has the same ability, or what its molecular mechanism might be. Rap1A activates PLCε in response to the stimulation of β-adrenergic receptors (β-ARs), translocating the complex to the perinuclear membrane. Because the C-terminal Ras association (RA2) domain of PLCε was proposed to the primary binding site for Rap1A, we first confirmed using purified proteins that the RA2 domain is indeed essential for activation by Rap1A. However, we also showed that the PLCε pleckstrin homology (PH) domain and first two EF hands (EF1/2) are required for Rap1A activation, and identified hydrophobic residues on the surface of the RA2 domain that are also necessary for activation. Finally, small angle X-ray scattering (SAXS) showed that Rap1A binding induces and stabilizes discrete conformational states in PLCε variants that can be activated by the GTPase. This data, together with the recent structure of a catalytically active fragment of PLCe, provide the first evidence that Rap1A, and by extension Ras, allosterically activate the lipase by promoting and stabilizing interactions between the RA2 domain and the PLC core.

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“…While this phospholipase contains a C2 domain that could direct the protein to the PM, a recent 3D structure of (almost) full length PLCE1 provided insights into its mechanisms and PM localization [52]. The model proposed that the C2 domain was not used for PM binding, but rather the αX-Y helix in the lipase domain.…”

Section: Plce1mentioning

confidence: 99%

The Ins and Outs of RAS Effector Complexes

Kiel¹,

Matallanas²,

Kölch³

2021

Preprint

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RAS oncogenes are amongst the most commonly mutated proteins in human cancers. They regulate a wide range of effector pathways that control cell proliferation, survival, differentiation, migration and metabolic status. Including aberrations in these pathways, RAS dependent signaling is altered in more than half of human cancers. Targeting mutant RAS proteins and their downstream oncogenic signaling pathways has been elusive. However, recent results comprising detailed molecular studies, large scale omics studies and computational modeling have painted a new and more comprehensive portrait of RAS signaling that helps us to understand the intricacies of RAS, how its physiological and pathophysiological functions are regulated, and how we can target them. Here, we review these efforts particularly trying to relate the detailed mechanistic studies with global functional studies. We highlight the importance of computational modeling and data integration to derive an actionable understanding of RAS signaling that will allow us to design new mechanism based therapies for RAS mutated cancers.

show abstract

Structure of phospholipase Cε reveals an integrated RA1 domain and previously unidentified regulatory elements

Cited by 13 publications

References 55 publications

The Ins and Outs of RAS Effector Complexes

The Ins and Outs of RAS Effector Complexes

Functional and structural characterization of allosteric activation of phospholipase Cε by Rap1A

The Ins and Outs of RAS Effector Complexes

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