title: Changes in lamina structure are followed by spatial reorganization of heterochromatic regions in caspase-8-activated human mesenchymal stem cells.
abstract: Apoptosis is fundamental to the regulation of homeostasis of stem cells in vivo. Whereas the pathways underlying the molecular and biochemical details of nuclear breakdown that accompanies apoptosis have been elucidated, the precise nature of nuclear reorganization that precedes the demolition phase is not fully understood. Here, we expressed an inducible caspase-8 in human mesenchymal stem cells, and quantitatively followed the early changes in nuclear organization during apoptosis. We found that caspase-8 induces alteration of the nuclear lamina and a subsequent spatial reorganization of both centromeres, which are shifted towards a peripheral localization, and telomeres, which form aggregates. This nuclear reorganization correlates with caspase-3 sensitivity of lamina proteins, because the expression of lamin mutant constructs with caspase-3 hypersensitivity resulted in a caspase-8-independent appearance of lamina intranuclear structures and telomere aggregates, whereas application of a caspase inhibitor restrains these changes in nuclear reorganization. Notably, upon activation of apoptosis, we observed no initial changes in the spatial organization of the promyelocytic leukemia nuclear bodies (PML-NBs). We suggest that during activation of the caspase-8 pathway changes in the lamina structure precede changes in heterochromatin spatial organization, and the subsequent breakdown of lamina and PML-NB.
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Einsteinweg 20, 2300RC Leiden, The Netherlands. v.raz@lumc.nl
authors.forename: Vered
authors.initials: V
authors.lastname: Raz
authors.forename: Françoise
authors.initials: F
authors.lastname: Carlotti
authors.forename: Bart J
authors.initials: BJ
authors.lastname: Vermolen
authors.forename: Egge
authors.initials: E
authors.lastname: van der Poel
authors.forename: Willem C R
authors.initials: WC
authors.lastname: Sloos
authors.forename: Shoshan
authors.initials: S
authors.lastname: Knaän-Shanzer
authors.forename: Antoine A F
authors.initials: AA
authors.lastname: de Vries
authors.forename: Rob C
authors.initials: RC
authors.lastname: Hoeben
authors.forename: Ian T
authors.initials: IT
authors.lastname: Young
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Tanke
authors.forename: Yuval
authors.initials: Y
authors.lastname: Garini
authors.forename: Roeland W
authors.initials: RW
authors.lastname: Dirks
mesh_terms: D015153:Blotting, Western; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D002503:Centromere / Q000378:metabolism; D004789:Enzyme Activation; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006570:Heterochromatin / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D034921:Lamin Type B / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009154:Mutation / Q000235:genetics; D034881:Nuclear Lamina / Q000378:metabolism*; D016615:Telomere / Q000378:metabolism
chemical_list: D006570:Heterochromatin; D034921:Lamin Type B; D049452:Green Fluorescent Proteins; D053181:Caspase 8