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Microinjection of antibodies targeting the lamin A/C histone-binding site blocks mitotic entry and reveals separate chromatin interactions with HP1, CenpB and PML

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Original languageEnglish
Article number9
JournalCells
Volume6
Issue number2
DOIs
StatePublished - 25 Mar 2017

Abstract

Lamins form a scaffold lining the nucleus that binds chromatin and contributes to spatial genome organization; however, due to the many other functions of lamins, studies knocking out or altering the lamin polymer cannot clearly distinguish between direct and indirect effects. To overcome this obstacle, we specifically targeted the mapped histone-binding site of A/C lamins by microinjecting antibodies specific to this region predicting that this would make the genome more mobile. No increase in chromatin mobility was observed; however, interestingly, injected cells failed to go through mitosis while control antibody-injected cells did. This effect was not due to crosslinking of the lamin polymer as Fab fragments also blocked mitosis. The lack of genome mobility suggested other lamin-chromatin interactions. To determine what these might be mini-lamin A constructs were expressed with or without the histone-binding site that assembled into independent intranuclear structures. HP1, CenpB and PML proteins accumulated at these structures for both constructs, indicating that other sites supporting chromatin interactions exist on lamin A. Together these results indicate that lamin A-chromatin interactions are highly redundant and more diverse than generally acknowledged and highlight the importance of trying to experimentally separate their individual functions.

    Research areas

  • lamin , cell cycle, genome stability, DNA replication, centromere proteins

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