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Hum Mol Genet. 2019 Jan 1;28(1):96-104. doi: 10.1093/hmg/ddy330.

GLS hyperactivity causes glutamate excess, infantile cataract and profound developmental delay.

Author information

1
Department of Genetics, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.
2
Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.
3
Department of Pediatrics, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.
4
Hubrecht Institute-KNAW, University Medical Center Utrecht, Utrecht University, Utrecht CT, The Netherlands.
5
Department of Radiology and Nuclear Medicine, VU University Medical Center, Amsterdam HV, The Netherlands.
6
Department of Radiology, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.
7
Department of Microbiology and Immunobiology, Harvard Medical School, Boston MA, USA.
8
Department of Ophthalmology, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.
9
Division of Evolution and Genomic Sciences, The University of Manchester, Manchester M139WL, UK.
10
Manchester Centre for Genomic Medicine, St Mary's Hospital, Manchester M139WL, UK.
11
Department of Medical Physiology, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.
12
Department of Child Neurology, VU University Medical Center, Amsterdam HV, The Netherlands.
13
Molecular Cancer Research, University Medical Center Utrecht, Utrecht University, Utrecht CX, The Netherlands.

Abstract

Loss-of-function mutations in glutaminase (GLS), the enzyme converting glutamine into glutamate, and the counteracting enzyme glutamine synthetase (GS) cause disturbed glutamate homeostasis and severe neonatal encephalopathy. We report a de novo Ser482Cys gain-of-function variant in GLS encoding GLS associated with profound developmental delay and infantile cataract. Functional analysis demonstrated that this variant causes hyperactivity and compensatory downregulation of GLS expression combined with upregulation of the counteracting enzyme GS, supporting pathogenicity. Ser482Cys-GLS likely improves the electrostatic environment of the GLS catalytic site, thereby intrinsically inducing hyperactivity. Alignment of +/-12.000 GLS protein sequences from >1000 genera revealed extreme conservation of Ser482 to the same degree as catalytic residues. Together with the hyperactivity, this indicates that Ser482 is evolutionarily preserved to achieve optimal-but submaximal-GLS activity. In line with GLS hyperactivity, increased glutamate and decreased glutamine concentrations were measured in urine and fibroblasts. In the brain (both grey and white matter), glutamate was also extremely high and glutamine was almost undetectable, demonstrated with magnetic resonance spectroscopic imaging at clinical field strength and subsequently supported at ultra-high field strength. Considering the neurotoxicity of glutamate when present in excess, the strikingly high glutamate concentrations measured in the brain provide an explanation for the developmental delay. Cataract, a known consequence of oxidative stress, was evoked in zebrafish expressing the hypermorphic Ser482Cys-GLS and could be alleviated by inhibition of GLS. The capacity to detoxify reactive oxygen species was reduced upon Ser482Cys-GLS expression, providing an explanation for cataract formation. In conclusion, we describe an inborn error of glutamate metabolism caused by a GLS hyperactivity variant, illustrating the importance of balanced GLS activity.

PMID:
30239721
DOI:
10.1093/hmg/ddy330
[Indexed for MEDLINE]

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