5 years ago

The WYL domain of the Thermotoga elfii PIF1 helicase is an accessory single-stranded DNA binding module

Sausen, Andis, M., Alladin, N. M., Bochman, C. W., A.
In the yeast Saccharomyces cerevisiae, the PIF1 family helicases Pif1 and Rrm3 aid in the maintenance of nuclear and mitochondrial genome stability. Despite great progress in understanding the roles of these enzymes as a whole, the functions of the N- and C-terminal domains that flank their central helicase core remain unclear. Here, we characterized the biochemical activities of the Thermotoga elfii PIF1 helicase (TePif1), which contains a C-terminal WYL domain. As is typical of helicases from thermophilic organisms, recombinant TePif1 was amenable to over-expression and purification, thermostable in vitro, and displayed activities similar to its better-studied eukaryotic homologs. We also found that the WYL domain was necessary for high affinity single-stranded DNA (ssDNA) binding and impacted both ATPase and helicase activities. Our results indicate that TePif1 and other bacterial PIF1 helicases may act as accessory helicases at replication forks to sense ssDNA levels behind the replicative helicase. Further, our findings predict that the domains of unknown function found in TePif1 homologs like S. cerevisiae Pif1 and Rrm3 may contain motifs needed for ssDNA binding.

Publisher URL: http://biorxiv.org/cgi/content/short/163188v1

DOI: 10.1101/163188

You might also like
Discover & Discuss Important Research

Keeping up-to-date with research can feel impossible, with papers being published faster than you'll ever be able to read them. That's where Researcher comes in: we're simplifying discovery and making important discussions happen. With over 19,000 sources, including peer-reviewed journals, preprints, blogs, universities, podcasts and Live events across 10 research areas, you'll never miss what's important to you. It's like social media, but better. Oh, and we should mention - it's free.

  • Download from Google Play
  • Download from App Store
  • Download from AppInChina

Researcher displays publicly available abstracts and doesn’t host any full article content. If the content is open access, we will direct clicks from the abstracts to the publisher website and display the PDF copy on our platform. Clicks to view the full text will be directed to the publisher website, where only users with subscriptions or access through their institution are able to view the full article.