Mutation of the Myxoma virus SERP2 P1-site to prevent proteinase inhibition causes apoptosis in cultured RK-13 cells and attenuates disease in rabbits, but mutation to alter specificity causes apoptosis without reducing virulence.
Myxoma virus (MYX) prevents apoptosis in RK-13 cells and forms thick dermal lesions with 100% mortality in rabbits. MYX encodes the virulence factor SERP2, a serine proteinase inhibitor (serpin). SERP2 was mutated to evaluate SERP2 function during MYX infection. MYXDeltaSERP2::lacZ (deleted for SERP2) did not inhibit apoptosis in RK-13 cells; infected rabbits had thin dermal lesions and <10% mortality. MYX-SERP2-D294A, a P1-site aspartate to alanine mutant, inactivated the serpin; infection was indistinguishable from MYXDeltaSERP2::lacZ. SERP2-D294E prevented inhibition of caspase-8, caspase-10 and granzyme-B; and MYX-SERP2-D294E failed to block apoptosis in RK-13 cells, but was fully virulent in rabbits. MYXDeltaSERP2::crmA expressed crmA instead of SERP2 and inhibited apoptosis in cell culture, but caused thin lesions and only 70% mortality in rabbits, hence crmA cannot fully substitute for SERP2. Control of apoptosis in culture does not correlate with virulence in rabbits. Virulence may instead depend on inhibition of proinflammatory proteinases by SERP2.
MacNeill
Amy L
AL
University of Illinois College of Veterinary Medicine, Department of Pathobiology, 2001 S. Lincoln Ave., Urbana, IL 61802, USA. almac@uiuc.edu <almac@uiuc.edu>
Turner
Peter C
PC
Moyer
Richard W
RW
D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D053804:Granzymes / Q000378:metabolism; D006801:Humans; D009154:Mutation; D009233:Myxoma virus / Q000235:genetics / Q000472:pathogenicity*; D009234:Myxomatosis, Infectious / Q000401:mortality / Q000503:physiopathology / Q000821:virology; D010447:Peptide Hydrolases / Q000378:metabolism; D011817:Rabbits; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D014774:Virulence
C120680:Serp2 protein, myxoma virus; D014764:Viral Proteins; D010447:Peptide Hydrolases; D053804:Granzymes