Comprehensive analysis of A-type lamin tail region for repairing the nuclear lamina

Kono, Y.; Pack, C.-G.; Ichikawa, T.; Komatsubara, A.; Adam, S. A.; Rolas, L.; Nourshargh, S.; Medalia, O.; Goldman, R. D.; Fukuma, T.; Kimura, H.; Shimi, T.

doi:10.1101/2023.09.02.555826

bioRxiv · 10.1101/2023.09.02.555826

Comprehensive analysis of A-type lamin tail region for repairing the nuclear lamina

Kono, Y.·Pack, C.-G.·Ichikawa, T.·Komatsubara, A.·Adam, S. A.·Rolas, L.·Nourshargh, S.·Medalia, O.·Goldman, R. D.·Fukuma, T.·Kimura, H.·Shimi, T.

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Abstract

The nuclear lamina (NL) lines the nuclear envelope (NE) to maintain nuclear structure in metazoan cells. The major NL components, the nuclear lamins contribute to the protection against NE rupture induced by mechanical stress. Lamin A (LA) and a short form of the splicing variant lamin C (LC) are diffused from the nucleoplasm to sites of NE rupture in immortalized mouse embryonic fibroblasts (MEFs). LA localization to the rupture sites is significantly slow and weak compared to LC because of its relatively small pool in the nucleoplasm, but the precise mechanism remains unknown. In this study, we induce NE rupture in wild-type and LA/C-knockout MEFs, and Hutchinson-Gilford Progeria syndrome (HGPS) knock-in MEFs that express progerin, a LA mutant lacking the second proteolytic cleavage site, by laser microirradiation and AFM indentation. The farnesylation at the CaaX motif of unprocessed LA and the inhibition of the second proteolytic cleavage decrease the nucleoplasmic pool and slow the localization to the rupture sites in a long-time window (60-70 min) after the induction of NE rupture. Our data could explain the defective repair of NE rupture in HGPS through the farnesylation at the CaaX motif of unprocessed progerin. In addition, unique segments in LA-specific tail region cooperate with each other to inhibit the rapid accumulation within a short-time window (3 min) that is also observed with LC. Significance StatementNuclear lamins are the major components of the nuclear lamina (NL) that lies the nuclear envelope (NE). Lamin A (LA) is slowly localized to sites of nuclear envelope (NE) rupture compared to lamin C (LC). This study reveals that the farnesylation at the CaaX motif of unprocessed LA and the inhibition of the second proteolytic cleavage decrease the nucleoplasmic pool and slow the localization to the rupture sites within a long-time window (60-70 min) after the induction of NE rupture, which could explain the defective repair of NE rupture in Hutchinson-Gilford Progeria syndrome (HGPS). Additionally, unique segments in LA-specific tail region are critical for inhibiting the rapid accumulation within a short-time window (3 min).

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Kono, Y., Pack, C.-G., Ichikawa, T., Komatsubara, A., Adam, S. A., Rolas, L., Nourshargh, S., Medalia, O., Goldman, R. D., Fukuma, T., Kimura, H., Shimi, T.. 2023-09-02. Comprehensive analysis of A-type lamin tail region for repairing the nuclear lamina. https://doi.org/10.1101/2023.09.02.555826

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