The Biochemical Activities of the Saccharomyces cerevisiae Pif1 Helicase Are Regulated by Its N-Terminal Domain
Molecular and Cellular Biochemistry Department, Indiana University, Bloomington, IN 47405, USA
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Author to whom correspondence should be addressed.
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These authors contributed equally to this work.
Genes 2019, 10(6), 411; https://doi.org/10.3390/genes10060411
Received: 31 March 2019 / Revised: 17 May 2019 / Accepted: 20 May 2019 / Published: 28 May 2019
(This article belongs to the Special Issue DNA Helicases: Mechanisms, Biological Pathways, and Disease Relevance)
Pif1 family helicases represent a highly conserved class of enzymes involved in multiple aspects of genome maintenance. Many Pif1 helicases are multi-domain proteins, but the functions of their non-helicase domains are poorly understood. Here, we characterized how the N-terminal domain (NTD) of the Saccharomyces cerevisiae Pif1 helicase affects its functions both in vivo and in vitro. Removal of the Pif1 NTD alleviated the toxicity associated with Pif1 overexpression in yeast. Biochemically, the N-terminally truncated Pif1 (Pif1ΔN) retained in vitro DNA binding, DNA unwinding, and telomerase regulation activities, but these activities differed markedly from those displayed by full-length recombinant Pif1. However, Pif1ΔN was still able to synergize with the Hrq1 helicase to inhibit telomerase activity in vitro, similar to full-length Pif1. These data impact our understanding of Pif1 helicase evolution and the roles of these enzymes in the maintenance of genome integrity.
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Keywords:
DNA helicase; Saccharomyces cerevisiae; Pif1; telomerase; telomere
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MDPI and ACS Style
Nickens, D.G.; Sausen, C.W.; Bochman, M.L. The Biochemical Activities of the Saccharomyces cerevisiae Pif1 Helicase Are Regulated by Its N-Terminal Domain. Genes 2019, 10, 411. https://doi.org/10.3390/genes10060411
AMA Style
Nickens DG, Sausen CW, Bochman ML. The Biochemical Activities of the Saccharomyces cerevisiae Pif1 Helicase Are Regulated by Its N-Terminal Domain. Genes. 2019; 10(6):411. https://doi.org/10.3390/genes10060411
Chicago/Turabian StyleNickens, David G.; Sausen, Christopher W.; Bochman, Matthew L. 2019. "The Biochemical Activities of the Saccharomyces cerevisiae Pif1 Helicase Are Regulated by Its N-Terminal Domain" Genes 10, no. 6: 411. https://doi.org/10.3390/genes10060411
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