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

Ridge Karen M; Stenvall Carl-Gustaf A; Heikkilä Taina R. H; Toivola Diana M.; Foisner Roland; Jansson Theresia; Nyström Joel H; Adam Stephen A; Butler-Hallissey Ciarán; Goldman Robert D

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

Ridge Karen M
Stenvall Carl-Gustaf A
Heikkilä Taina R. H
Toivola Diana M.
Foisner Roland
Jansson Theresia
Nyström Joel H
Adam Stephen A
Butler-Hallissey Ciarán
Goldman Robert D
Katso/Avaa
mbc.e20-06-0387.pdf (2.570Mb)
Lataukset: 

American Society for Cell Biology
doi:10.1091/mbc.E20-06-0387
URI
https://doi.org/10.1091/mbc.E20-06-0387
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Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe2022112967841
Tiivistelmä

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.

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