SPEG interacts with myotubularin, and its deficiency causes centronuclear myopathy with dilated cardiomyopathy

  • Am J Hum Genet. 2014 Aug 7;95(2):218-26. doi: 10.1016/j.ajhg.2014.07.004.
Pankaj B Agrawal  1 Christopher R Pierson  2 Mugdha Joshi  3 Xiaoli Liu  4 Gianina Ravenscroft  5 Behzad Moghadaszadeh  3 Tiffany Talabere  6 Marissa Viola  7 Lindsay C Swanson  3 Göknur Haliloğlu  8 Beril Talim  9 Kyle S Yau  5 Richard J N Allcock  10 Nigel G Laing  5 Mark A Perrella  11 Alan H Beggs  12
Affiliations
  • 1. Division of Genetics and Genomics, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA; Division of Newborn Medicine, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA; Manton Center for Orphan Disease Research, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA. Electronic address: [email protected].
  • 2. Department of Pathology and Laboratory Medicine, Nationwide Children's Hospital and the Ohio State University College of Medicine, Columbus, OH 43205, USA.
  • 3. Division of Genetics and Genomics, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA; Manton Center for Orphan Disease Research, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
  • 4. Department of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston MA 02115, USA.
  • 5. Harry Perkins Institute of Medical Research and the Centre for Medical Research, University of Western Australia, Nedlands, WA 6009, Australia.
  • 6. Research Institute, Nationwide Children's Hospital and the Ohio State University College of Medicine, Columbus, OH 43205, USA.
  • 7. Division of Genetics and Genomics, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
  • 8. Neurology Unit, Department of Pediatrics, Hacettepe University Children's Hospital, Ankara 06100, Turkey.
  • 9. Pathology Unit, Department of Pediatrics, Hacettepe University Children's Hospital, Ankara 06100, Turkey.
  • 10. Lotterywest State Biomedical Facility Genomics and School of Pathology and Laboratory Medicine, University of Western Australia, Perth, WA 6009, Australia.
  • 11. Department of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston MA 02115, USA; Department of Newborn Medicine, Brigham and Women's Hospital, Boston MA 02115, USA.
  • 12. Division of Genetics and Genomics, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA; Manton Center for Orphan Disease Research, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA. Electronic address: [email protected].
Abstract

Centronuclear myopathies (CNMs) are characterized by muscle weakness and increased numbers of central nuclei within myofibers. X-linked myotubular myopathy, the most common severe form of CNM, is caused by mutations in MTM1, encoding myotubularin (MTM1), a lipid Phosphatase. To increase our understanding of MTM1 function, we conducted a yeast two-hybrid screen to identify MTM1-interacting proteins. Striated muscle preferentially expressed protein kinase (SPEG), the product of SPEG complex locus (SPEG), was identified as an MTM1-interacting protein, confirmed by immunoprecipitation and immunofluorescence studies. SPEG knockout has been previously associated with severe dilated cardiomyopathy in a mouse model. Using whole-exome Sequencing, we identified three unrelated CNM-affected probands, including two with documented dilated cardiomyopathy, carrying homozygous or compound-heterozygous SPEG mutations. SPEG was markedly reduced or absent in two individuals whose muscle was available for immunofluorescence and immunoblot studies. Examination of muscle samples from Speg-knockout mice revealed an increased frequency of central nuclei, as seen in human subjects. SPEG localizes in a double line, flanking desmin over the Z lines, and is apparently in alignment with the terminal cisternae of the sarcoplasmic reticulum. Examination of human and murine MTM1-deficient muscles revealed similar abnormalities in staining patterns for both desmin and SPEG. Our results suggest that mutations in SPEG, encoding SPEG, cause a CNM phenotype as a result of its interaction with MTM1. SPEG is present in cardiac muscle, where it plays a critical role; therefore, individuals with SPEG mutations additionally present with dilated cardiomyopathy.