title: Proliferating cell nuclear antigen prevents trinucleotide repeat expansions by promoting repeat deletion and hairpin removal.
abstract: DNA base lesions and base excision repair (BER) within trinucleotide repeat (TNR) tracts modulate repeat instability through the coordination among the key BER enzymes DNA polymerase β, flap endonuclease 1 (FEN1) and DNA ligase I (LIG I). However, it remains unknown whether BER cofactors can also alter TNR stability. In this study, we discovered that proliferating cell nuclear antigen (PCNA), a cofactor of BER, promoted CAG repeat deletion and removal of a CAG repeat hairpin during BER in a duplex CAG repeat tract and CAG hairpin loop, respectively. We showed that PCNA stimulated LIG I activity on a nick across a small template loop during BER in a duplex (CAG)20repeat tract promoting small repeat deletions. Surprisingly, we found that during BER in a hairpin loop, PCNA promoted reannealing of the upstream flap of a double-flap intermediate, thereby facilitating the formation of a downstream flap and stimulating FEN1 cleavage activity and hairpin removal. Our results indicate that PCNA plays a critical role in preventing CAG repeat expansions by modulating the structures of dynamic DNA via cooperation with BER enzymes. We provide the first evidence that PCNA prevents CAG repeat expansions during BER by promoting CAG repeat deletion and removal of a TNR hairpin.
authors: Biochemistry Ph.D. Program, Florida International University, 11200 SW 8th Street, Miami, FL 33199, United States.
authors: Jill M
authors: JM
authors: Beaver
authors: Department of Chemistry and Biochemistry, Florida International University, 11200 SW 8th Street, Miami, FL 33199, United States.
authors: Yanhao
authors: Y
authors: Lai
authors: Department of Chemistry and Biochemistry, Florida International University, 11200 SW 8th Street, Miami, FL 33199, United States.
authors: Shantell J
authors: SJ
authors: Rolle
authors: Biochemistry Ph.D. Program, Florida International University, 11200 SW 8th Street, Miami, FL 33199, United States; Department of Chemistry and Biochemistry, Florida International University, 11200 SW 8th Street, Miami, FL 33199, United States; Biomolecular Sciences Institute, School of Integrated Sciences and Humanities, Florida International University, 11200 SW 8th Street, Miami, FL 33199, United States. Electronic address: yualiu@fiu.edu.
authors: Yuan
authors: Y
authors: Liu
mesh_terms: D001483:Base Sequence; D004249:DNA Damage; D000072481:DNA Ligase ATP / Q000235:genetics* / Q000378:metabolism; D019951:DNA Polymerase beta / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000235:genetics / Q000378:metabolism; D045585:Flap Endonucleases / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D009690:Nucleic Acid Conformation; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion; D019680:Trinucleotide Repeat Expansion; D018911:Trinucleotide Repeats
chemical_list: C000607892:LIG1 protein, human; D018809:Proliferating Cell Nuclear Antigen; D019951:DNA Polymerase beta; D045585:Flap Endonucleases; C468317:FEN1 protein, human; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D000072481:DNA Ligase ATP
keywords: Base excision repair (BER); DNA polymerase β (pol β); DNA repair; Flap endonuclease 1 (FEN1); Proliferating cell nuclear antigen (PCNA); Trinucleotide repeats