α-Fodrin is required for the organization of functional microtubules during mitosis

Cell Cycle. 2019 Oct;18(20):2713-2726. doi: 10.1080/15384101.2019.1656476. Epub 2019 Aug 27.

Abstract

The cytoskeleton protein α-fodrin plays a major role in maintaining structural stability of membranes. It was also identified as part of the brain γ-tubulin ring complex, the major microtubule nucleator. Here, we investigated the requirement of α-fodrin for microtubule spindle assembly during mitotic progression. We found that α-fodrin depletion results in abnormal mitosis with uncongressed chromosomes, leading to prolonged activation of the spindle assembly checkpoint and a severe mitotic delay. Further, α-fodrin repression led to the formation of shortened spindles with unstable kinetochore-microtubule attachments. We also found that the mitotic kinesin CENP-E had reduced levels at kinetochores to likely account for the chromosome misalignment defects in α-fodrin-depleted cells. Importantly, we showed these cells to exhibit reduced levels of detyrosinated α-tubulin, which primarily drives CENP-E localization. Since proper microtubule dynamics and chromosome alignment are required for completion of normal mitosis, this study reveals an unforeseen role of α-fodrin in regulating mitotic progression. Future studies on these lines of observations should reveal important mechanistic insight for fodrin's involvement in cancer.

Keywords: CENP-E; SAC; detyrosinated tubulin; mitosis; spindle; α-Fodrin.

MeSH terms

  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Line, Tumor
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Chromosome Segregation
  • Humans
  • Kinetochores / metabolism
  • M Phase Cell Cycle Checkpoints / genetics*
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Microtubules / metabolism*
  • Mitosis / genetics*
  • RNA, Small Interfering
  • Spindle Apparatus / metabolism
  • Tubulin / metabolism
  • Tyrosine / metabolism

Substances

  • Carrier Proteins
  • Chromosomal Proteins, Non-Histone
  • Microfilament Proteins
  • RNA, Small Interfering
  • Tubulin
  • centromere protein E
  • fodrin
  • Tyrosine

Grants and funding

RKN, JSS and DD got fellowships from University Grants Commission, India. RKN was also funded by European Molecular Biology Organization short term fellowship (No. 7103). [IF/00916/2014]; Rajiv Gandhi Centre for Biotechnology, Government of India; North Regional Operational Program [NORTE-01-0145-FEDER-000029]; Fundação para a Ciência e a Tecnologia (PT) [PTDC/BIA-CEL/31120/2017]; North Regional Operational Program [NORTE-01-0145-FEDER-000029]; Fundação para a Ciência e Tecnologia grant [PTDC/ BEX- BCM/1921/ 2014]; DST-SERB [EMR/2015/001676]; Fundação para a Ciência e a Tecnologia (PT) [PTDC/BEX-BCM/2090/2014].