ORF157 from the archaeal virus Acidianus filamentous virus 1 defines a new class of nuclease.
Acidianus filamentous virus 1 (AFV1) (Lipothrixviridae) is an enveloped filamentous virus that was characterized from a crenarchaeal host. It infects Acidianus species that thrive in the acidic hot springs (>85 degrees C and pH <3) of Yellowstone National Park, WY. The AFV1 20.8-kb, linear, double-stranded DNA genome encodes 40 putative open reading frames whose sequences generally show little similarity to other genes in the sequence databases. Because three-dimensional structures are more conserved than sequences and hence are more effective at revealing function, we set out to determine protein structures from putative AFV1 open reading frames (ORF). The crystal structure of ORF157 reveals an alpha+beta protein with a novel fold that remotely resembles the nucleotidyltransferase topology. In vitro, AFV1-157 displays a nuclease activity on linear double-stranded DNA. Alanine substitution mutations demonstrated that E86 is essential to catalysis. AFV1-157 represents a novel class of nuclease, but its exact role in vivo remains to be determined.
Goulet
Adeline
A
Architecture et Fonction des Macromolcéules Biologiques, CNRS and Universités d'Aix-Marseille I & II, Marseille, France.
Pina
Mery
M
Redder
Peter
P
Prangishvili
David
D
Vera
Laura
L
Lichière
Julie
J
Leulliot
Nicolas
N
van Tilbeurgh
Herman
H
Ortiz-Lombardia
Miguel
M
Campanacci
Valérie
V
Cambillau
Christian
C
D041541:Acidianus / Q000821:virology*; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D018360:Crystallography, X-Ray; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D045482:Hot Springs / Q000382:microbiology; D023643:Lipothrixviridae / Q000737:chemistry* / Q000201:enzymology* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D014764:Viral Proteins; D004247:DNA; D003851:Deoxyribonucleases