Cytoplasmic keratins couple with and maintain nuclear envelope integrity in colonic epithelial cells

Mol Biol Cell. 2022 Nov 1;33(13):ar121. doi: 10.1091/mbc.E20-06-0387. Epub 2022 Aug 24.

Abstract

Keratin intermediate filaments convey mechanical stability and protection against stress to epithelial cells. Keratins are essential for colon health, as seen in keratin 8 knockout (K8-/-) mice exhibiting a colitis phenotype. We hypothesized that keratins support the nuclear envelope and lamina in colonocytes. K8-/- colonocytes in vivo exhibit significantly decreased levels of lamins A/C, B1, and B2 in a colon-specific and cell-intrinsic manner. CRISPR/Cas9- or siRNA-mediated K8 knockdown in Caco-2 cells similarly decreased lamin levels, which recovered after reexpression of K8 following siRNA treatment. Nuclear area was not decreased, and roundness was only marginally increased in cells without K8. Down-regulation of K8 in adult K8flox/flox;Villin-CreERt2 mice following tamoxifen administration significantly decreased lamin levels at day 4 when K8 levels had reduced to 40%. K8 loss also led to reduced levels of plectin, LINC complex, and lamin-associated proteins. While keratins were not seen in the nucleoplasm without or with leptomycin B treatment, keratins were found intimately located at the nuclear envelope and complexed with SUN2 and lamin A. Furthermore, K8 loss in Caco-2 cells compromised nuclear membrane integrity basally and after shear stress. In conclusion, colonocyte K8 helps maintain nuclear envelope and lamina composition and contributes to nuclear integrity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caco-2 Cells
  • Colon / metabolism
  • Cytoskeletal Proteins / metabolism
  • Epithelial Cells / metabolism
  • Humans
  • Keratin-8* / genetics
  • Keratins* / metabolism
  • Lamin Type A / metabolism
  • Mice
  • Nuclear Envelope / metabolism
  • Plectin / metabolism
  • RNA, Small Interfering / metabolism
  • Tamoxifen

Substances

  • Cytoskeletal Proteins
  • Keratin-8
  • Lamin Type A
  • Plectin
  • RNA, Small Interfering
  • Tamoxifen
  • Keratins