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- W2890176597 abstract "The linker of nucleoskeleton and cytoskeleton (LINC) is a conserved nuclear envelope-spanning molecular bridge that is responsible for the mechanical integration of the nucleus with the cytoskeleton. LINC complexes are formed by a transluminal interaction between the outer and inner nuclear membrane KASH and SUN proteins, respectively. Despite recent structural insights, our mechanistic understanding of LINC complex assembly remains limited by the lack of an experimental system for its in vitro reconstitution and manipulation. Here, we describe artificial nuclear membranes (ANMs) as a synthetic biology platform based on mammalian cell-free expression for the rapid reconstitution of SUN proteins in supported lipid bilayers. We demonstrate that SUN1 and SUN2 are oriented in ANMs with solvent-exposed C-terminal KASH-binding SUN domains. We also find that SUN2 possesses a single transmembrane domain, while SUN1 possesses three. Finally, SUN protein-containing ANMs bind synthetic KASH peptides, thereby reconstituting the LINC complex core. This work represents the first in vitro reconstitution of KASH-binding SUN proteins in supported lipid bilayers using cell-free expression, which will be invaluable for testing proposed models of LINC complex assembly and its regulation." @default.
- W2890176597 created "2018-09-27" @default.
- W2890176597 creator A5029393245 @default.
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- W2890176597 creator A5043839270 @default.
- W2890176597 creator A5058661506 @default.
- W2890176597 creator A5088989964 @default.
- W2890176597 date "2018-01-01" @default.
- W2890176597 modified "2023-09-27" @default.
- W2890176597 title "A synthetic biology platform for the reconstitution and mechanistic dissection of LINC complex assembly" @default.
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- W2890176597 doi "https://doi.org/10.1242/jcs.219451" @default.
- W2890176597 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/30262467" @default.
- W2890176597 hasPublicationYear "2018" @default.
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