The splicing factor CCAR1 regulates the Fanconi anemia/BRCA pathway

Mol Cell. 2024 Jul 25;84(14):2618-2633.e10. doi: 10.1016/j.molcel.2024.06.031. Epub 2024 Jul 17.

Abstract

The twenty-three Fanconi anemia (FA) proteins cooperate in the FA/BRCA pathway to repair DNA interstrand cross-links (ICLs). The cell division cycle and apoptosis regulator 1 (CCAR1) protein is also a regulator of ICL repair, though its possible function in the FA/BRCA pathway remains unknown. Here, we demonstrate that CCAR1 plays a unique upstream role in the FA/BRCA pathway and is required for FANCA protein expression in human cells. Interestingly, CCAR1 co-immunoprecipitates with FANCA pre-mRNA and is required for FANCA mRNA processing. Loss of CCAR1 results in retention of a poison exon in the FANCA transcript, thereby leading to reduced FANCA protein expression. A unique domain of CCAR1, the EF hand domain, is required for interaction with the U2AF heterodimer of the spliceosome and for excision of the poison exon. Taken together, CCAR1 is a splicing modulator required for normal splicing of the FANCA mRNA and other mRNAs involved in various cellular pathways.

Keywords: CCAR1; DNA repair; EF hand; FANCA; Fanconi anemia; U2AF1/2; alternative splicing; poison exon.

MeSH terms

  • Apoptosis Regulatory Proteins* / genetics
  • Apoptosis Regulatory Proteins* / metabolism
  • BRCA1 Protein / genetics
  • BRCA1 Protein / metabolism
  • BRCA2 Protein / genetics
  • BRCA2 Protein / metabolism
  • Cell Cycle Proteins* / genetics
  • Cell Cycle Proteins* / metabolism
  • DNA Repair
  • Endodeoxyribonucleases
  • Exons
  • Fanconi Anemia Complementation Group A Protein* / genetics
  • Fanconi Anemia Complementation Group A Protein* / metabolism
  • Fanconi Anemia* / genetics
  • Fanconi Anemia* / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Protein Binding
  • RNA Precursors / genetics
  • RNA Precursors / metabolism
  • RNA Splicing*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Spliceosomes / genetics
  • Spliceosomes / metabolism
  • Splicing Factor U2AF* / genetics
  • Splicing Factor U2AF* / metabolism

Substances

  • BRCA1 Protein
  • BRCA1 protein, human
  • BRCA2 Protein
  • BRCA2 protein, human
  • Endodeoxyribonucleases
  • FANCA protein, human
  • Fanconi Anemia Complementation Group A Protein
  • RBBP8 protein, human
  • RNA Precursors
  • RNA, Messenger
  • Splicing Factor U2AF
  • U2AF1 protein, human
  • CCAR1 protein, human
  • Cell Cycle Proteins
  • Apoptosis Regulatory Proteins