DNA substrate recognition and processing by the full-length human UPF1 helicase

• Published in: Nucleic acids research Vol. 45; no. 12; pp. 7354 - 7366
• Main Authors: Dehghani-Tafti, Saba, Sanders, Cyril M
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 07.07.2017
• Subjects: Base Sequence; Cloning, Molecular; DNA - chemistry; DNA - genetics; DNA - metabolism; DNA Replication; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Humans; Nonsense Mediated mRNA Decay; Nucleic Acid Conformation; Nucleic Acid Enzymes; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA Stability; RNA, Messenger - chemistry; RNA, Messenger - genetics; RNA, Messenger - metabolism; Substrate Specificity; Trans-Activators - genetics; Trans-Activators - metabolism
• DOI: 10.1093/nar/gkx478
• ISSN: 0305-1048

UPF1 is a conserved helicase required for nonsense-mediated decay (NMD) regulating mRNA stability in the cytoplasm. Human UPF1 (hUPF1) is also needed for nuclear DNA replication. While loss of NMD is tolerated, loss of hUPF1 induces a DNA damage response and cell cycle arrest. We have analysed nucleic acid (NA) binding and processing by full-length hUPF1. hUPF1 unwinds non-B and B-form DNA and RNA substrates in vitro. Unlike many helicases involved in genome stability no hUPF1 binding to DNA structures stabilized by inter-base-pair hydrogen bonding was observed. Alternatively, hUPF1 binds to single-stranded NAs (ssNA) with apparent affinity increasing with substrate length and with no preference for binding RNA or DNA or purine compared to pyrimidine polynucleotides. However, the data show a pronounced nucleobase bias with a preference for binding poly (U) or d(T) while d(A) polymers bind with low affinity. Although the data indicate that hUPF1 must bind a ssNA segments to initiate unwinding they also raise the possibility that hUPF1 has significantly reduced affinity for ssNA structures with stacked bases. Overall, the NA processing activities of hUPF1 are consistent with its function in mRNA regulation and suggest that roles in DNA replication could also be influenced by base sequence.