Antagonistic roles of canonical and Alternative-RPA in disease-associated tandem CAG repeat instability

Cell. 2023 Oct 26;186(22):4898-4919.e25. doi: 10.1016/j.cell.2023.09.008. Epub 2023 Oct 11.

Abstract

Expansions of repeat DNA tracts cause >70 diseases, and ongoing expansions in brains exacerbate disease. During expansion mutations, single-stranded DNAs (ssDNAs) form slipped-DNAs. We find the ssDNA-binding complexes canonical replication protein A (RPA1, RPA2, and RPA3) and Alternative-RPA (RPA1, RPA3, and primate-specific RPA4) are upregulated in Huntington disease and spinocerebellar ataxia type 1 (SCA1) patient brains. Protein interactomes of RPA and Alt-RPA reveal unique and shared partners, including modifiers of CAG instability and disease presentation. RPA enhances in vitro melting, FAN1 excision, and repair of slipped-CAGs and protects against CAG expansions in human cells. RPA overexpression in SCA1 mouse brains ablates expansions, coincident with decreased ATXN1 aggregation, reduced brain DNA damage, improved neuron morphology, and rescued motor phenotypes. In contrast, Alt-RPA inhibits melting, FAN1 excision, and repair of slipped-CAGs and promotes CAG expansions. These findings suggest a functional interplay between the two RPAs where Alt-RPA may antagonistically offset RPA's suppression of disease-associated repeat expansions, which may extend to other DNA processes.

Keywords: Alternative replication protein A (Alt-RPA); BioID protein interactome; DNA repair (FAN1, MSH2, MSH3, MSH6, HTT); Huntington's disease (HD); RPA1, RPA2, RPA3, RPA4; Replication protein A (RPA); Spinocerebellar ataxia type 1 (SCA1); slipped-DNA; tandem repeat expansions; trinucleotide CAG repeat expansions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • DNA / genetics
  • DNA Mismatch Repair
  • Humans
  • Huntington Disease / genetics
  • Mice
  • Proteins / genetics
  • Replication Protein A* / metabolism
  • Spinocerebellar Ataxias / genetics
  • Trinucleotide Repeat Expansion*

Substances

  • DNA
  • Proteins
  • Replication Protein A