Novel Families of Archaeo-Eukaryotic Primases Associated with Mobile Genetic Elements of Bacteria and Archaea

J Mol Biol. 2018 Mar 2;430(5):737-750. doi: 10.1016/j.jmb.2017.11.014. Epub 2017 Dec 11.


Cellular organisms in different domains of life employ structurally unrelated, non-homologous DNA primases for synthesis of a primer for DNA replication. Archaea and eukaryotes encode enzymes of the archaeo-eukaryotic primase (AEP) superfamily, whereas bacteria uniformly use primases of the DnaG family. However, AEP genes are widespread in bacterial genomes raising questions regarding their provenance and function. Here, using an archaeal primase-polymerase PolpTN2 encoded by pTN2 plasmid as a seed for sequence similarity searches, we recovered over 800 AEP homologs from bacteria belonging to 12 highly diverse phyla. These sequences formed a supergroup, PrimPol-PV1, and could be classified into five novel AEP families which are characterized by a conserved motif containing an arginine residue likely to be involved in nucleotide binding. Functional assays confirm the essentiality of this motif for catalytic activity of the PolpTN2 primase-polymerase. Further analyses showed that bacterial AEPs display a range of domain organizations and uncovered several candidates for novel families of helicases. Furthermore, sequence and structure comparisons suggest that PriCT-1 and PriCT-2 domains frequently fused to the AEP domains are related to each other as well as to the non-catalytic, large subunit of archaeal and eukaryotic primases, and to the recently discovered PriX subunit of archaeal primases. Finally, genomic neighborhood analysis indicates that the identified AEPs encoded in bacterial genomes are nearly exclusively associated with highly diverse integrated mobile genetic elements, including integrative conjugative plasmids and prophages.

Keywords: DNA replication; Thermococcus plasmids; evolution; helicases; structural modeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Archaea / enzymology*
  • Archaea / genetics*
  • Bacteria / enzymology
  • Bacteria / genetics*
  • Catalytic Domain
  • DNA Helicases / metabolism
  • DNA Primase / classification
  • DNA Primase / metabolism*
  • DNA Replication
  • Eukaryota / enzymology*
  • Eukaryota / genetics*
  • Evolution, Molecular
  • Interspersed Repetitive Sequences
  • Models, Molecular
  • Protein Conformation
  • Protein Domains
  • Sequence Alignment
  • Thermococcus / genetics


  • DNA Primase
  • DNA Helicases