title: Regulation of the MID1 protein function is fine-tuned by a complex pattern of alternative splicing.
abstract: Clinical features of Opitz BBB/G syndrome are confined to defects of the developing ventral midline, whereas the causative gene, MID1, is ubiquitously expressed. Therefore, a non-redundant physiological function of the MID1 product appears to be developmentally restricted. Here, we report the identification of several alternative MID1 exons in human, mouse and fugu. We show that splice variants of the MID1 gene that are comparable in terms of function occur in the three organisms, suggesting an important role in the regulation of the MID1 protein function. Accordingly, we observed differential MID1 transcript patterns in a tissue-specific manner by Northern blot and RT-PCR. The identified splice variants cause loss-of-function effects via several mechanisms. Some introduce a stop codon followed by a novel poly(A(+)) tail, leading to the formation of C-terminally truncated proteins. Dominant negative effects through altered binding to the MID1-interacting protein alpha4 in vitro could be demonstrated in a couple of cases. Others carry premature termination codons without poly(A(+)) tails. These are degraded by nonsense mediated mRNA decay (NMD). Our data reveal a mechanism conserved in human, mouse and fugu that regulates developmentally restricted MID1 activity and suggest NMD to be critical in the translational regulation of a ubiquitously transcribed mRNA.
authors.affiliation: Max-Planck Institute for Molecular Genetics, Ihnestrasse 63-73, 14195 Berlin, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Winter
authors.forename: Tanja
authors.initials: T
authors.lastname: Lehmann
authors.forename: Sybille
authors.initials: S
authors.lastname: Krauss
authors.forename: Alexander
authors.initials: A
authors.lastname: Trockenbacher
authors.forename: Zofia
authors.initials: Z
authors.lastname: Kijas
authors.forename: John
authors.initials: J
authors.lastname: Foerster
authors.forename: Vanessa
authors.initials: V
authors.lastname: Suckow
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Yaspo
authors.forename: Andreas
authors.initials: A
authors.lastname: Kulozik
authors.forename: Vera
authors.initials: V
authors.lastname: Kalscheuer
authors.forename: Rainer
authors.initials: R
authors.lastname: Schneider
authors.forename: Susann
authors.initials: S
authors.lastname: Schweiger
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D018389:Codon, Nonsense / Q000235:genetics; D005091:Exons / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D008868:Microtubule Proteins / Q000235:genetics* / Q000502:physiology; D009687:Nuclear Proteins / Q000235:genetics* / Q000502:physiology; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D023741:Takifugu / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics* / Q000502:physiology; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases
chemical_list: D018389:Codon, Nonsense; D008868:Microtubule Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; C109052:MID1 protein, human; D044767:Ubiquitin-Protein Ligases