Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA.
Functions of prokaryotic Argonautes (pAgo) have long remained elusive. Recently, Argonautes of the bacteria Rhodobacter sphaeroides and Thermus thermophilus were demonstrated to be involved in host defense. The Argonaute of the archaeon Pyrococcus furiosus (PfAgo) belongs to a different branch in the phylogenetic tree, which is most closely related to that of RNA interference-mediating eukaryotic Argonautes. Here we describe a functional and mechanistic characterization of PfAgo. Like the bacterial counterparts, archaeal PfAgo contributes to host defense by interfering with the uptake of plasmid DNA. PfAgo utilizes small 5'-phosphorylated DNA guides to cleave both single stranded and double stranded DNA targets, and does not utilize RNA as guide or target. Thus, with respect to function and specificity, the archaeal PfAgo resembles bacterial Argonautes much more than eukaryotic Argonautes. These findings demonstrate that the role of Argonautes is conserved through the bacterial and archaeal domains of life and suggests that eukaryotic Argonautes are derived from DNA-guided DNA-interfering host defense systems.
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, 6703 HB Wageningen, The Netherlands.
Daan C
http://orcid.org/0000-0003-4412-9191
DC
Swarts
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, 6703 HB Wageningen, The Netherlands.
Jorrit W
JW
Hegge
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, 6703 HB Wageningen, The Netherlands.
Ismael
I
Hinojo
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
Masami
M
Shiimori
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
Michael A
MA
Ellis
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
Justin
J
Dumrongkulraksa
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
Rebecca M
RM
Terns
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
Michael P
http://orcid.org/0000-0001-8330-1789
MP
Terns
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, 6703 HB Wageningen, The Netherlands john.vanderoost@wur.nl.
John
J
van der Oost
D019843:Archaeal Proteins / Q000737:chemistry / Q000378:metabolism*; D060565:Argonaute Proteins / Q000737:chemistry / Q000378:metabolism*; D020134:Catalytic Domain; D004247:DNA / Q000378:metabolism; D053837:DNA Cleavage; D004706:Endodeoxyribonucleases / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D020027:Pyrococcus furiosus / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic
D019843:Archaeal Proteins; D060565:Argonaute Proteins; D004247:DNA; D004706:Endodeoxyribonucleases