A trans-acting protein effect causes severe eye malformation in the Mp mouse

PLoS Genet. 2013;9(12):e1003998. doi: 10.1371/journal.pgen.1003998. Epub 2013 Dec 12.

Abstract

Mp is an irradiation-induced mouse mutation associated with microphthalmia, micropinna and hind limb syndactyly. We show that Mp is caused by a 660 kb balanced inversion on chromosome 18 producing reciprocal 3-prime gene fusion events involving Fbn2 and Isoc1. The Isoc1-Fbn2 fusion gene (Isoc1(Mp)) mRNA has a frameshift and early stop codon resulting in nonsense mediated decay. Homozygous deletions of Isoc1 do not support a significant developmental role for this gene. The Fbn2-Isoc1 fusion gene (Fbn2 (Mp)) predicted protein consists of the N-terminal Fibrillin-2 (amino acids 1-2646, exons 1-62) lacking the C-terminal furin-cleavage site with a short out-of-frame extension encoded by the final exon of Isoc1. The Mp limb phenotype is consistent with that reported in Fbn2 null embryos. However, severe eye malformations, a defining feature of Mp, are not seen in Fbn2 null animals. Fibrillin-2(Mp) forms large fibrillar structures within the rough endoplasmic reticulum (rER) associated with an unfolded protein response and quantitative mass spectrometry shows a generalised defect in protein secretion in conditioned media from mutant cells. In the embryonic eye Fbn2 is expressed within the peripheral ciliary margin (CM). Mp embryos show reduced canonical Wnt-signalling in the CM - known to be essential for ciliary body development - and show subsequent aplasia of CM-derived structures. We propose that the Mp "worse-than-null" eye phenotype plausibly results from a failure in normal trafficking of proteins that are co-expressed with Fbn2 within the CM. The prediction of similar trans-acting protein effects will be an important challenge in the medical interpretation of human mutations from whole exome sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromosome Inversion / genetics
  • Chromosomes, Human, Pair 18 / genetics
  • Exons
  • Eye / growth & development
  • Eye / physiopathology
  • Eye Abnormalities / genetics*
  • Eye Abnormalities / physiopathology
  • Fibrillin-2
  • Fibrillins
  • Frameshift Mutation
  • Humans
  • Mice
  • Microfilament Proteins / genetics*
  • Microphthalmos / genetics*
  • Microphthalmos / physiopathology
  • Mutation / radiation effects*
  • Phenotype
  • Syndactyly / genetics
  • Syndactyly / physiopathology
  • Wnt Signaling Pathway / genetics

Substances

  • FBN2 protein, human
  • Fbn2 protein, mouse
  • Fibrillin-2
  • Fibrillins
  • Microfilament Proteins