title: An activated 5' cryptic splice site in the human ALG3 gene generates a premature termination codon insensitive to nonsense-mediated mRNA decay in a new case of congenital disorder of glycosylation type Id (CDG-Id).
abstract: A defect of the dolichyl-P-Man:Man5GlcNAc2-PP-dolichyl mannosyltransferase encoded by the ALG3 gene (alias NOT56L) causes congenital disorder of glycosylation type Id (CDG-Id). In this work, a new mutation in the ALG3 gene causing atypical splicing is described with characterization of expression levels and transcript stabilities of the different splice products. A silent mutation in exon 1 of the ALG3 gene (c.165C<T) resulted in a deletion in the corresponding transcripts (c.160_196del) due to the activation of a cryptic donor splice site. Expression studies revealed that negligible amounts of normal transcripts were present in the patient. The deletion in the ALG3 gene generated a premature termination codon (PTC) coding for an ALG3 protein truncated after the first N-terminal transmembranous domain (p.Val54fsX66). Nonsense mediated decay (NMD) of mRNA is a general mechanism for clearing of RNA molecules containing suitable PTCs. However, suppression of NMD using cycloheximide had no influence on ALG3 transcript levels, although the PTCs of the transcript fulfill the criteria for the initiation of NMD. The results presented in this work demonstrate that factors abrogating NMD of the ALG3 gene exists and that the ALG3 gene can serve as a valuable tool for further investigations of the regulation of NMD.
authors.affiliation: Department of Pediatrics, University Hospital of Münster, Münster, Germany. deneckj@uni-muenster.de
authors.forename: Jonas
authors.initials: J
authors.lastname: Denecke
authors.forename: Christian
authors.initials: C
authors.lastname: Kranz
authors.forename: Dirk
authors.initials: D
authors.lastname: Kemming
authors.forename: Hans-Georg
authors.initials: HG
authors.lastname: Koch
authors.forename: Thorsten
authors.initials: T
authors.lastname: Marquardt
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002239:Carbohydrate Metabolism, Inborn Errors / Q000235:genetics* / Q000378:metabolism; D018389:Codon, Nonsense; D018388:Codon, Terminator; D003513:Cycloheximide / Q000494:pharmacology; D004252:DNA Mutational Analysis; D005091:Exons; D006031:Glycosylation; D006801:Humans; D008364:Mannosyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D012322:RNA Precursors / Q000378:metabolism; D022821:RNA Splice Sites; D012326:RNA Splicing; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: D018389:Codon, Nonsense; D018388:Codon, Terminator; D012322:RNA Precursors; D022821:RNA Splice Sites; D012333:RNA, Messenger; D003513:Cycloheximide; C489947:ALG3 protein, human; D008364:Mannosyltransferases; C066968:dolichyl-P-Man:Man(5)GlcNAc(2)-PP-dolichol alpha-1,3-mannosyltransferase