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

Terence Gall-Duncan; Jennifer Luo; Carla-Marie Jurkovic; Laura A Fischer; Kyota Fujita; Amit L Deshmukh; Rachel J Harding; Stephanie Tran; Mustafa Mehkary; Vanessa Li; David E Leib; Ran Chen; Hikari Tanaka; Amanda G Mason; Dominique Lévesque; Mahreen Khan; Mortezaali Razzaghi; Tanya Prasolava; Stella Lanni; Nozomu Sato; Marie-Christine Caron; Gagan B Panigrahi; Peixiang Wang; Rachel Lau; Arturo López Castel; Jean-Yves Masson; Lynette Tippett; Clinton Turner; Maria Spies; Albert R La Spada; Eric I Campos; Maurice A Curtis; François-Michel Boisvert; Richard L M Faull; Beverly L Davidson; Masayuki Nakamori; Hitoshi Okazawa; Marc S Wold; Christopher E Pearson

doi:10.1016/j.cell.2023.09.008

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.

Authors

Terence Gall-Duncan¹, Jennifer Luo¹, Carla-Marie Jurkovic², Laura A Fischer³, Kyota Fujita⁴, Amit L Deshmukh⁵, Rachel J Harding⁶, Stephanie Tran¹, Mustafa Mehkary¹, Vanessa Li¹, David E Leib⁷, Ran Chen⁸, Hikari Tanaka⁴, Amanda G Mason⁹, Dominique Lévesque², Mahreen Khan¹, Mortezaali Razzaghi¹⁰, Tanya Prasolava⁵, Stella Lanni⁵, Nozomu Sato⁵, Marie-Christine Caron¹¹, Gagan B Panigrahi⁵, Peixiang Wang⁵, Rachel Lau⁵, Arturo López Castel¹², Jean-Yves Masson¹¹, Lynette Tippett¹³, Clinton Turner¹⁴, Maria Spies¹⁰, Albert R La Spada¹⁵, Eric I Campos¹, Maurice A Curtis¹⁶, François-Michel Boisvert², Richard L M Faull¹⁶, Beverly L Davidson⁷, Masayuki Nakamori¹⁷, Hitoshi Okazawa⁴, Marc S Wold¹⁰, Christopher E Pearson¹⁸

Affiliations

¹ Genetics & Genome Biology, The Hospital for Sick Children, Toronto, ON, Canada; Molecular Genetics, University of Toronto, Toronto, ON, Canada.
² Immunology and Cell Biology, Université de Sherbrooke, Sherbrooke, QC, Canada.
³ Developmental Biology and Center of Regenerative Medicine, Washington University School of Medicine, St. Louis, MO, USA.
⁴ Neuropathology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
⁵ Genetics & Genome Biology, The Hospital for Sick Children, Toronto, ON, Canada.
⁶ Structural Genomics Consortium, University of Toronto, Toronto, ON M5G 1L7, Canada; Pharmacology and Toxicology, University of Toronto, Toronto, ON M5S 1A8, Canada.
⁷ Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19146, USA.
⁸ Pediatrics, Division of Hematology and Oncology, Washington University School of Medicine, St. Louis, MO 63110, USA.
⁹ Human Genetics, Leiden University Medical Center, Leiden, the Netherlands.
¹⁰ Biochemistry and Molecular Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA.
¹¹ CHU de Québec-Université Laval, Oncology Division, Molecular Biology, Medical Biochemistry, and Pathology, Laval University Cancer Research Center, Québec, QC, Canada.
¹² BIOTECMED, University of Valencia, Valencia, Spain.
¹³ School of Psychology, University of Auckland, Auckland, New Zealand; University Research Centre for Brain Research, University of Auckland, Auckland, New Zealand.
¹⁴ Anatomical Pathology, LabPlus, Auckland City Hospital, Auckland, New Zealand.
¹⁵ Pathology & Laboratory Medicine, Neurology, and Biological Chemistry, University of California, Irvine School of Medicine, Irvine, CA, USA; Neurobiology & Behavior, University of California, Irvine, Irvine, CA, USA; Center for Neurotherapeutics, University of California, Irvine, Irvine, CA 92697, USA.
¹⁶ University Research Centre for Brain Research, University of Auckland, Auckland, New Zealand; Anatomy and Medical Imaging, University of Auckland, Auckland, New Zealand.
¹⁷ Neurology, Osaka University Graduate School of Medicine, Osaka, Japan.
¹⁸ Genetics & Genome Biology, The Hospital for Sick Children, Toronto, ON, Canada; Structural Genomics Consortium, University of Toronto, Toronto, ON M5G 1L7, Canada. Electronic address: cepearson.sickkids@gmail.com.

PMID: 37827155
DOI: 10.1016/j.cell.2023.09.008

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

Abstract

Publication types

MeSH terms

Substances

Grants and funding