CRISPR screens in sister chromatid cohesion defective cells reveal PAXIP1-PAGR1 as regulator of chromatin association of cohesin

Janne J.M. van Schie, Klaas de Lint, Thom M. Molenaar, Macarena Moronta Gines, Jesper A. Balk, Martin A. Rooimans, Khashayar Roohollahi, Govind M. Pai, Lauri Borghuis, Anisha R. Ramadhin, Francesco Corazza, Josephine C. Dorsman, Kerstin S. Wendt, Rob M.F. Wolthuis, Job de Lange

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The cohesin complex regulates higher order chromosome architecture through maintaining sister chromatid cohesion and folding chromatin by DNA loop extrusion. Impaired cohesin function underlies a heterogeneous group of genetic syndromes and is associated with cancer. Here, we mapped the genetic dependencies of human cell lines defective of cohesion regulators DDX11 and ESCO2. The obtained synthetic lethality networks are strongly enriched for genes involved in DNA replication and mitosis and support the existence of parallel sister chromatid cohesion establishment pathways. Among the hits, we identify the chromatin binding, BRCT-domain containing protein PAXIP1 as a novel cohesin regulator. Depletion of PAXIP1 severely aggravates cohesion defects in ESCO2 mutant cells, leading to mitotic cell death. PAXIP1 promotes global chromatin association of cohesin, independent of DNA replication, a function that cannot be explained by indirect effects of PAXIP1 on transcription or DNA repair. Cohesin regulation by PAXIP1 requires its binding partner PAGR1 and a conserved FDF motif in PAGR1. PAXIP1 co-localizes with cohesin on multiple genomic loci, including active gene promoters and enhancers. Possibly, this newly identified role of PAXIP1-PAGR1 in regulating cohesin occupancy on chromatin is also relevant for previously described functions of PAXIP1 in transcription, immune cell maturation and DNA repair.

Original languageEnglish
Pages (from-to)9594-9609
Number of pages16
JournalNucleic Acids Research
Volume51
Issue number18
DOIs
Publication statusPublished - 13 Oct 2023

Cite this