A novel homozygous splicing mutation of CASC5 causes primary microcephaly in a large Pakistani family

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• 作者：Sandra Szczepanski ; Muhammad Sajid Hussain ; Ilknur Sur ; Janine Altmüller…
• 刊名：Human Genetics
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：135
• 期：2
• 页码：157-170
• 全文大小：4,773 KB
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• 作者单位：Sandra Szczepanski (1) (2) (3) (6)
  Muhammad Sajid Hussain (1) (2) (3) (6)
  Ilknur Sur (2) (3) (6)
  Janine Altmüller (1) (4)
  Holger Thiele (1)
  Uzma Abdullah (5)
  Syeda Seema Waseem (5)
  Abubakar Moawia (5)
  Gudrun Nürnberg (1)
  Angelika Anna Noegel (1) (2) (3) (6)
  Shahid Mahmood Baig (5)
  Peter Nürnberg (1) (3) (6)
  
  1. Cologne Center for Genomics (CCG), University of Cologne, 50931, Cologne, Germany
  2. Institute of Biochemistry I, Medical Faculty, University of Cologne, Joseph-Stelzmann-Str. 52, 50931, Cologne, Germany
  3. Center for Molecular Medicine Cologne (CMMC), University of Cologne, Weyertal 115b, 50931, Cologne, Germany
  6. Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, 50931, Cologne, Germany
  4. Institute of Human Genetics, University of Cologne, 50931, Cologne, Germany
  5. Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Molecular Medicine
  Internal Medicine
  Metabolic Diseases
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1203

文摘

Primary microcephaly is a disorder characterized by a small head and brain associated with impaired cognitive capabilities. Mutations in 13 different genes encoding centrosomal proteins and cell cycle regulators have been reported to cause the disease. CASC5, a gene encoding a protein important for kinetochore formation and proper chromosome segregation during mitosis, has been suggested to be associated with primary microcephaly-4 (MCPH4). This was based on one mutation only and circumstantial functional evidence. By combining homozygosity mapping and whole-exome sequencing in an MCPH family from Pakistan, we identified a second mutation (NM_170589.4;c.6673-19T>A) in CASC5. This mutation induced skipping of exon 25 of CASC5 resulting in a frameshift and the introduction of a premature stop codon (p.Met2225Ilefs*7). The C-terminally truncated protein lacks 118 amino acids that encompass the region responsible for the interaction with the hMIS12 complex, which is essential for proper chromosome alignment and segregation. Furthermore, we showed a down-regulation of CASC5 mRNA and reduction of the amount of CASC5 protein by quantitative RT-PCR and western blot analysis, respectively. As a further sign of functional deficits, we observed dispersed dots of CASC5 immunoreactive material outside the metaphase plate of dividing patient fibroblasts. Normally, CASC5 is a component of the kinetochore of metaphase chromosomes. A higher mitotic index in patient cells indicated a mitotic arrest in the cells carrying the mutation. We also observed lobulated and fragmented nuclei as well as micronuclei in the patient cells. Moreover, we detected an altered DNA damage response with higher levels of γH2AX and 53BP1 in mutant as compared to control fibroblasts. Our findings substantiate the proposed role of CASC5 for primary microcephaly and suggest that it also might be relevant for genome stability. S. Szczepanski and M. S. Hussain are joint first authors.Electronic supplementary materialThe online version of this article (doi:10.​1007/​s00439-015-1619-5) contains supplementary material, which is available to authorized users.