Effects of the interactions of classical swine fever virus Core protein with proteins of the SUMOylation pathway on virulence in swine.
Here we have identified host cell proteins involved with the cellular SUMOylation pathway, SUMO-1 (small ubiquitin-like modifier) and UBC9, a SUMO-1 conjugating enzyme that interact with classical swine fever virus (CSFV) Core protein. Five highly conserved lysine residues (K179, K180, K220, K221, and K246) within the CSFV Core were identified as putative SUMOylation sites. Analysis of these interactions showed that K179A, K180A, and K221A substitutions disrupt Core-SUMO-1 binding, while K220A substitution precludes Core-UBC9 binding. In vivo, Core mutant viruses (K179A, K180A, K220A, K221A) and (K220A, K221A) harboring those substitutions were attenuated in swine. These data shows a clear correlation between the disruption of Core protein binding to SUMO-1 and UBC9 and CSFV attenuation. Overall, these data suggest that the interaction of Core with the cellular SUMOylation pathway plays a significant role in the CSFV growth cycle in vivo.
Gladue
D P
DP
Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. douglas.gladue@ars.usda.gov
Holinka
L G
LG
Fernandez-Sainz
I J
IJ
Prarat
M V
MV
O'Donell
V
V
Vepkhvadze
N
N
Lu
Z
Z
Rogers
K
K
Risatti
G R
GR
Borca
M V
MV
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002460:Cell Line; D006691:Classical Swine Fever / Q000473:pathology / Q000821:virology; D006692:Classical Swine Fever Virus / Q000472:pathogenicity*; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D025842:SUMO-1 Protein / Q000378:metabolism*; D016415:Sequence Alignment; D013552:Swine; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism*; D014758:Viral Core Proteins / Q000378:metabolism*; D014774:Virulence
D050505:Mutant Proteins; D025842:SUMO-1 Protein; D014758:Viral Core Proteins; D044763:Ubiquitin-Conjugating Enzymes; D008239:Lysine