Miro-dependent mitochondrial pool of CENP-F and its farnesylated C-terminal domain are dispensable for normal development in mice

PLoS Genet. 2019 Mar 11;15(3):e1008050. doi: 10.1371/journal.pgen.1008050. eCollection 2019 Mar.


CENP-F is a large, microtubule-binding protein that regulates multiple cellular processes including chromosome segregation and mitochondrial trafficking at cytokinesis. This multiplicity of functions is mediated through the binding of various partners, like Bub1 at the kinetochore and Miro at mitochondria. Due to the multifunctionality of CENP-F, the cellular phenotypes observed upon its depletion are difficult to interpret and there is a need to genetically separate its different functions by preventing binding to selected partners. Here we engineer a CENP-F point-mutant that is deficient in Miro binding and thus is unable to localize to mitochondria, but retains other localizations. We introduce this mutation in cultured human cells using CRISPR/Cas9 system and show it causes a defect in mitochondrial spreading similar to that observed upon Miro depletion. We further create a mouse model carrying this CENP-F variant, as well as truncated CENP-F mutants lacking the farnesylated C-terminus of the protein. Importantly, one of these truncations leads to ~80% downregulation of CENP-F expression. We observe that, despite the phenotypes apparent in cultured cells, mutant mice develop normally. Taken together, these mice will serve as important models to study CENP-F biology at organismal level. In addition, because truncations of CENP-F in humans cause a lethal disease termed Strømme syndrome, they might also be relevant disease models.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • CRISPR-Cas Systems
  • Cell Line
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Eye Abnormalities / genetics
  • Humans
  • Intestinal Atresia / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred DBA
  • Mice, Transgenic
  • Microcephaly / genetics
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Mitochondria / metabolism
  • Mitochondrial Proteins / chemistry
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism*
  • Point Mutation
  • Prenylation
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • rho GTP-Binding Proteins / chemistry
  • rho GTP-Binding Proteins / genetics
  • rho GTP-Binding Proteins / metabolism*


  • Chromosomal Proteins, Non-Histone
  • Microfilament Proteins
  • Miro-1 protein, mouse
  • Mitochondrial Proteins
  • centromere protein F
  • Rhot2 protein, mouse
  • rho GTP-Binding Proteins

Supplementary concepts

  • Jejunal Atresia with Microcephaly and Ocular Anomalies

Grants and funding

This work was funded by grants from the ERC (337906-Orga-Net) and the Swiss National Science Foundation (PP00P3_13365) to BK. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.