Identification of two novel beta-mannosidosis-associated sequence variants: biochemical analysis of beta-mannosidase (MANBA) missense mutations.
Beta-mannosidosis (OMIM # 248510) is an autosomal-recessive lysosomal storage disorder caused by deficiency of the lysosomal enzyme beta-mannosidase (MANBA, E.C. 3.2.1.25). The disorder has been reported in goat, cattle and man. The human disorder is rare and only 20 cases in 16 families have been reported. We have sequenced the exons and exon-intron borders in a European patient with infantile onset of beta-mannosidosis. The patient was compound heterozygous for a silent mutation (c.375A>G) in exon 3 causing alternative splicing, and a missense mutation (c.1513T>C, p.Ser505Pro) in exon 12. The alternative splicing event deleted four nucleotides from the transcript and was predicted to result in premature termination of translation. In order to evaluate the consequence of the missense mutation, we inserted the human beta-mannosidase gene into an expression vector, performed site-directed mutagenesis and expressed the normal and mutant enzyme in COS-7 cells. We also included the previously reported beta-mannosidosis-associated missense mutations c.544C>T (p.Arg182Trp) and c.1175G>A (p.Gly392Glu), which were found in patients presenting a milder phenotype. Cells transfected with the wild-type construct showed a 33-fold increase in beta-mannosidase activity compared to mock-transfected cells, whereas cells transfected with the mutant constructs showed no detectable increase in activity. We propose that the milder phenotype described in some beta-mannosidosis patients with missense mutations in the MANBA gene is not due to residual beta-mannosidase activity, but rather caused by epigenetic and/or environmental factors.
Riise Stensland
Hilde Monica Frostad
HMF
Department of Medical Genetics, University Hospital of Northern Norway, Tromsø, Norway.
Persichetti
Emanuele
E
Department of Internal Medicine, University of Perugia, Perugia, Italy.
Sorriso
Carmelita
C
Department of Internal Medicine, University of Perugia, Perugia, Italy.
Hansen
Gaute Martin
GM
Department of Medical Biochemistry, University of Tromsø, Tromsø, Norway.
Bibi
Lucia
L
Department of Internal Medicine, University of Perugia, Perugia, Italy.
Paciotti
Silvia
S
Department of Internal Medicine, University of Perugia, Perugia, Italy.
Balducci
Chiara
C
Department of Internal Medicine, University of Perugia, Perugia, Italy.
Beccari
Tommaso
T
Department of Internal Medicine, University of Perugia, Perugia, Italy.
D017398:Alternative Splicing; D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D018389:Codon, Nonsense; D004252:DNA Mutational Analysis; D015870:Gene Expression; D006801:Humans; D020125:Mutation, Missense; D044465:White People / Q000235:genetics; D044902:beta-Mannosidase / Q000235:genetics* / Q000378:metabolism; D044905:beta-Mannosidosis / Q000201:enzymology* / Q000235:genetics
D018389:Codon, Nonsense; D044902:beta-Mannosidase