A novel splice site FUS mutation in a familial ALS case: effects on protein expression.
Amyotroph Lateral Scler Frontotemporal Degener
; 23(1-2): 128-136, 2022 02.
Article
en En
| MEDLINE
| ID: mdl-33879000
ABSTRACT
Objective:
To investigate the impact of a novel heterozygous FUS mutation in the acceptor splice site of intron 14 (c.1542 - 1 g > t) on protein expression in Peripheral Blood Mononuclear Cells (PBMC) from a familial ALS patient.Methods:
PBMC were isolated for mRNA analysis (cDNA synthesis, sequencing and one-step RT-PCR), Western Immunoblot (WI), and Immunofluorescence (IF).Results:
cDNA analysis revealed the skipping of exon 15 and a premature stop codon at c.228. RT-PCR showed reduced FUS mRNA by more than half compared to a healthy control (HC) and an ALS patient without genetic mutations (wtALS). In WI FUS band intensity in the proband was 30-50% compared to HC and wtALS. An antibody expected to detect only the wild-type protein did not reveal any reduction of FUS band intensity compared to the other antibodies. IF showed no difference among HC, wtALS, and the proband.Discussion:
The reduction of FUS mRNA and protein in PBMC suggests the absence of the truncated protein, probably due to nonsense-mediated decay, leading to loss of function.Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Leucocitos Mononucleares
/
Esclerosis Amiotrófica Lateral
Límite:
Adult
/
Humans
/
Male
/
Middle aged
Idioma:
En
Revista:
Amyotroph Lateral Scler Frontotemporal Degener
Año:
2022
Tipo del documento:
Article
País de afiliación:
Italia