PT  - JOURNAL ARTICLE
AU  - Mark Kittisopikul
AU  - Takeshi Shimi
AU  - Meltem Tatli
AU  - Joseph R. Tran
AU  - Yixian Zheng
AU  - Ohad Medalia
AU  - Khuloud Jaqaman
AU  - Stephen A. Adam
AU  - Robert D. Goldman
TI  - Computational analysis of lamin isoform interactions with nuclear pore complexes
AID  - 10.1101/2020.04.03.022798
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.04.03.022798
4099  - http://biorxiv.org/content/early/2020/07/16/2020.04.03.022798.short
4100  - http://biorxiv.org/content/early/2020/07/16/2020.04.03.022798.full
AB  - Nuclear lamin isoforms form fibrous meshworks associated with nuclear pore complexes (NPCs). Using data sets prepared from sub-pixel and segmentation analyses of 3D-Structured Illumination Microscopy images of WT and lamin isoform knockout mouse embryo fibroblasts, we determined with high precision the spatial association of NPCs with specific lamin isoform fibers. These relationships are retained in the enlarged lamin meshworks of Lmna-/- and Lmnb1-/- fibroblast nuclei. Cryo-ET observations reveal that the lamin filaments composing the fibers contact the nucleoplasmic ring of NPCs. Knockdown of the ring-associated nucleoporin ELYS induces NPC clusters that exclude lamin A/C fibers, but include LB1 and LB2 fibers. Knockdown of the nucleoporins TPR or NUP153 alter the arrangement of lamin fibers and NPCs. Evidence that the number of NPCs is regulated by specific lamin isoforms is presented. Overall the results demonstrate that lamin isoforms and nucleoporins act together to maintain the normal organization of lamin meshworks and NPCs within the nuclear envelope.Competing Interest StatementThe authors have declared no competing interest.