Nonstructural protein σ1s mediates reovirus-induced cell cycle arrest and apoptosis.
Reovirus nonstructural protein σ1s is implicated in cell cycle arrest at the G2/M boundary and induction of apoptosis. However, the contribution of σ1s to these effects in an otherwise isogenic viral background has not been defined. To evaluate the role of σ1s in cell cycle arrest and apoptosis, we used reverse genetics to generate a σ1s-null reovirus. Following infection with wild-type virus, we observed an increase in the percentage of cells in G2/M, whereas the proportion of cells in G2/M following infection with the σ1s-null mutant was unaffected. Similarly, we found that the wild-type virus induced substantially greater levels of apoptosis than the σ1s-null mutant. These data indicate that σ1s is required for both reovirus-induced cell cycle arrest and apoptosis. To define sequences in σ1s that mediate these effects, we engineered viruses encoding C-terminal σ1s truncations by introducing stop codons in the σ1s open reading frame. We also generated viruses in which charged residues near the σ1s amino terminus were replaced individually or as a cluster with nonpolar residues. Analysis of these mutants revealed that amino acids 1 to 59 and the amino-terminal basic cluster are required for induction of both cell cycle arrest and apoptosis. Remarkably, viruses that fail to induce cell cycle arrest and apoptosis also are attenuated in vivo. Thus, identical sequences in σ1s are required for reovirus-induced cell cycle arrest, apoptosis, and pathogenesis. Collectively, these findings provide evidence that the σ1s-mediated properties are genetically linked and suggest that these effects are mechanistically related.
Department of Microbiology and Immunology.
Karl W
KW
Boehme
Katharina
K
Hammer
William C
WC
Tollefson
Jennifer L
JL
Konopka-Anstadt
Takeshi
T
Kobayashi
Terence S
TS
Dermody
D020816:Amino Acid Motifs; D000818:Animals; D017209:Apoptosis; D059447:Cell Cycle Checkpoints; D002460:Cell Line; D006801:Humans; D012089:Mammalian orthoreovirus 3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D012088:Reoviridae Infections / Q000503:physiopathology* / Q000821:virology*; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D017361:Viral Nonstructural Proteins; C071901:sigma 1NS protein, Reovirus