A Novel Homozygous p.L539F Mutation Identified in<i>PINK1</i>Gene in a Moroccan Patient with Parkinsonism

Haj, Rafiqua Ben El; Regragui, W.; Tazi-Ahnini, Rachid; Skalli, A.; Bouslam, Naïma; Benomar, Ali; Yahyaoui, M.; Bouhouche, Ahmed

doi:10.1155/2016/3460234

Cited by 9 publications

(6 citation statements)

References 24 publications

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“…Under these assay conditions, WT PINK1 that was localized to mitochondria efficiently phosphorylates K63-Ub 4 ; however, this was completely abolished in mitochondria isolated from KI PINK1 and 3A PINK1-expressing cells ( figure 4 e ). Overall these studies demonstrate the critical role of the CTE hydrophobic residues in PINK1 activation and strongly suggest that the L539F mutant is not pathogenic and causal of PD in the case reported [ 28 ].…”

Section: Resultsmentioning

confidence: 65%

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Mapping of a N-terminal α-helix domain required for human PINK1 stabilization, Serine228 autophosphorylation and activation in cells

et al. 2022

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Autosomal recessive mutations in the PINK1 gene are causal for Parkinson's disease (PD). PINK1 encodes a mitochondrial localized protein kinase that is a master-regulator of mitochondrial quality control pathways. Structural studies to date have elaborated the mechanism of how mutations located within the kinase domain disrupt PINK1 function; however, the molecular mechanism of PINK1 mutations located upstream and downstream of the kinase domain is unknown. We have employed mutagenesis studies to define the minimal region of human PINK1 required for optimal ubiquitin phosphorylation, beginning at residue Ile111. Inspection of the AlphaFold human PINK1 structure model predicts a conserved N-terminal α-helical extension (NTE) domain forming an intramolecular interaction with the C-terminal extension (CTE), which we corroborate using hydrogen/deuterium exchange mass spectrometry of recombinant insect PINK1 protein. Cell-based analysis of human PINK1 reveals that PD-associated mutations (e.g. Q126P), located within the NTE : CTE interface, markedly inhibit stabilization of PINK1; autophosphorylation at Serine228 (Ser228) and Ubiquitin Serine65 (Ser65) phosphorylation. Furthermore, we provide evidence that NTE and CTE domain mutants disrupt PINK1 stabilization at the mitochondrial Translocase of outer membrane complex. The clinical relevance of our findings is supported by the demonstration of defective stabilization and activation of endogenous PINK1 in human fibroblasts of a patient with early-onset PD due to homozygous PINK1 Q126P mutations. Overall, we define a functional role of the NTE : CTE interface towards PINK1 stabilization and activation and show that loss of NTE : CTE interactions is a major mechanism of PINK1-associated mutations linked to PD.

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Section: Resultsmentioning

confidence: 65%

“…Of relevance is the location of residues mutated in PD including Cys125 and Gln126 within this interface as well as Glu117. The key residues in the α K helix mediating the interaction include Val528, Leu532 and Leu539 ( figure 3 c – e ) in which a PD case has been reported with homozygous Leu539Phe mutation [ 28 ].…”

Section: Resultsmentioning

confidence: 99%

Mapping of a N-terminal α-helix domain required for human PINK1 stabilization, Serine228 autophosphorylation and activation in cells

et al. 2022

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“…We did not observe any biochemical defect of the L539F mutation on PINK1 activation (Figure 4). This mutation has been reported in only one patient/family with PD compared to the other mutations we assessed which have been found in multiple independent families, and overall our data indicates that this mutation is not pathogenic and unlikely to be the cause of Parkinson’s in the reported case [30]. In contrast the clinical relevance and importance of the function of the NTE:CTE interface is supported by the demonstration of defective endogenous PINK1 stabilisation and activation in patient-derived fibroblasts of a homozygous Q126P mutation.…”

Section: Discussionmentioning

confidence: 59%

“…Under these assay conditions, WT PINK1 that was localised to mitochondria efficiently phosphorylates K63-Ub4, however, this was completely abolished in mitochondria isolated from KI PINK1 and 3A PINK1-expressing cells (Figure 4E). Overall these studies demonstrate the critical role of the CTE hydrophobic residues in PINK1 activation and strongly suggest that the L539F mutant is not pathogenic and causal of PD in the case reported [30].…”

Section: Biochemical Analysis Indicates Critical Role Of Nte:cte Interface For Pink1 Activation In Cellsmentioning

confidence: 65%

Mapping of a N-terminal α-helix domain required for human PINK1 stabilisation, Serine228 autophosphorylation and activation in cells

Kakade

Ojha

Raimi

et al. 2021

Preprint

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PINK1 encodes a mitochondrial localised protein kinase that is a master-regulator of mitochondrial quality control pathways. Structural studies to date have elaborated the mechanism of how mutations located within the kinase domain disrupt PINK1 function, however, the molecular mechanism of PINK1 mutations located upstream and downstream of the kinase domain are unknown. We have employed mutagenesis studies of human PINK1 in cells to define the minimal region of PINK1, required for optimal ubiquitin phosphorylation, beginning at residue Ile111. Bioinformatic analysis of the region spanning Ile111 to the kinase domain and inspection of the AlphaFold human PINK1 structure model predicts a conserved N-terminal alpha-helical domain extension (NTE domain) within this region corroborated by hydrogen/deuterium exchange mass spectrometry (HDX-MS) of recombinant insect PINK1 protein. The AlphaFold structure also predicts the NTE domain forms an intramolecular interaction with the C-terminal extension (CTE). Cell-based analysis of human PINK1 reveals that PD-associated mutations (e.g. Q126P), located within the NTE:CTE interface, markedly inhibit stabilization of PINK1; autophosphorylation at Serine228 (Ser228); and Ubiquitin Serine65 (Ser65) phosphorylation. Furthermore, we provide evidence that NTE domain mutants do not affect intrinsic catalytic kinase activity but do disrupt PINK1 stabilisation at the mitochondrial Translocase of outer membrane (TOM) complex. The clinical relevance of our findings is supported by the demonstration of defective stabilization and activation of endogenous PINK1 in human fibroblasts of a patient with early-onset PD due to homozygous PINK1 Q126P mutations. Overall, we define a functional role of the NTE:CTE interface towards PINK1 stabilisation and activation and show that loss of NTE:CTE interactions is a major mechanism of PINK1-associated mutations linked to PD.

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“…Indeed, two patients without PRKN CNV showed ROH at the 1p36 locus and had mutations in the PINK1 gene. These mutations consisted of the already known Q456X mutation in exon 7 and a new L539F variant in exon 8; the latter of which is located in the C-terminal domain of the PINK1 protein that we have already shown to be probably pathogenic ( 37 ).…”

Section: Discussionmentioning

confidence: 99%

Mutation Analysis of Consanguineous Moroccan Patients with Parkinson’s Disease Combining Microarray and Gene Panel

et al. 2017

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During the last two decades, 15 different genes have been reported to be responsible for the monogenic form of Parkinson’s disease (PD), representing a worldwide frequency of 5–10%. Among them, 10 genes have been associated with autosomal recessive PD, with PRKN and PINK1 being the most frequent. In a cohort of 145 unrelated Moroccan PD patients enrolled since 2013, 19 patients were born from a consanguineous marriage, of which 15 were isolated cases and 4 familial. One patient was homozygous for the common LRRK2 G2019S mutation and the 18 others who did not carry this mutation were screened for exon rearrangements in the PRKN gene using Affymetrix Cytoscan HD microarray. Two patients were determined homozygous for PRKN exon-deletions, while another patient presented with compound heterozygous inheritance (3/18, 17%). Two other patients showed a region of homozygosity covering the 1p36.12 locus and were sequenced for the candidate PINK1 gene, which revealed two homozygous point mutations: the known Q456X mutation in exon 7 and a novel L539F variation in exon 8. The 13 remaining patients were subjected to next-generation sequencing (NGS) that targeted a panel of 22 PD-causing genes and overlapping phenotypes. NGS data showed that two unrelated consanguineous patients with juvenile-onset PD (12 and 13 years) carried the same homozygous stop mutation W258X in the ATP13A2 gene, possibly resulting from a founder effect; and one patient with late onset (76 years) carried a novel heterozygous frameshift mutation in SYNJ1. Clinical analysis showed that patients with the ATP13A2 mutation developed juvenile-onset PD with a severe phenotype, whereas patients having either PRKN or PINK1 mutations displayed early-onset PD with a relatively mild phenotype. By identifying pathogenic mutations in 45% (8/18) of our consanguineous Moroccan PD series, we demonstrate that the combination of chromosomal microarray analysis and NGS is a powerful approach to pinpoint the genetic bases of autosomal recessive PD, particularly in countries with a high rate of consanguinity.

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A Novel Homozygous p.L539F Mutation Identified inPINK1Gene in a Moroccan Patient with Parkinsonism

Cited by 9 publications

References 24 publications

Mapping of a N-terminal α-helix domain required for human PINK1 stabilization, Serine228 autophosphorylation and activation in cells

Mapping of a N-terminal α-helix domain required for human PINK1 stabilization, Serine228 autophosphorylation and activation in cells

Mapping of a N-terminal α-helix domain required for human PINK1 stabilisation, Serine228 autophosphorylation and activation in cells

Mutation Analysis of Consanguineous Moroccan Patients with Parkinson’s Disease Combining Microarray and Gene Panel

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