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bioRxiv · 10.1101/2022.03.17.484746

Structure and functional mapping of the KRAB-KAP1 repressor complex

Stoll, G. A.·Pandiloski, N.·Douse, C. H.·Modis, Y.
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Abstract

Transposable elements (TEs) are a genetic reservoir from which new genes and regulatory elements can emerge. Expression of TEs can be pathogenic, however, and is tightly regulated. KRAB domain-containing zinc finger proteins (KRAB-ZFPs) recruit co-repressor KRAB-associated protein 1 (KAP1/TRIM28) to regulate many TEs but how KRAB-ZFPs and KAP1 interact remains unclear. Here, we report the crystal structure of the KAP1 tripartite motif (TRIM) in complex with the KRAB domain from a human KRAB-ZFP, ZNF93. Structure-guided mutations in the KAP1-KRAB binding interface abolished repressive activity in an epigenetic TE silencing assay. Deposition of H3K9me3 over thousands of loci is lost genome-wide in cells expressing a KAP1 variant with mutations that abolish KRAB binding. Our work identifies and functionally validates the KRAB-KAP1 molecular interface, which forms the nexus of a transcriptional control axis vital to vertebrates and underpins programmable gene repression by CRISPRi.

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Stoll, G. A., Pandiloski, N., Douse, C. H., Modis, Y.. 2022-03-17. Structure and functional mapping of the KRAB-KAP1 repressor complex. https://doi.org/10.1101/2022.03.17.484746

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