Non-human Primate Schlafen11 Inhibits Production of Both Host and Viral Proteins
Fig 6
Sites under positive selection do not govern species-specific differences in activity of Schlafen11.
(A) Plasmids encoding HIV-Gag-Pol, HIV-Rev, GFP, and eGFP were cotransfected into 293T cells along with a plasmid encoding the indicated great ape Schlafen11, or Chlor as a negative control. Cell lysates were subject to immunoblotting. (B) A phylogeny of the great apes is shown, with the activity of the Schlafen11 of each species indicated. Those in green have strong ability to suppress protein production when expressed in human cells, while those in red are weaker. (C) The seven amino acid positions that differ between chimpanzee and bonobo Schlafen11 are illustrated on a protein schematic (see S3 Fig for an alignment). Asterisks indicate sites under positive selection. The table shows the posterior probability of each site being under positive selection (PAML), as well as which sites were determined to be under positive selection by the other three tests. We liberally included position 345 in this group because it was detected in our PAML analysis with a posterior probability of >0.5, providing only weak support for selection (i.e. there is a 54% chance that this codon was correctly assigned to the dN/dS > 1 site class by PAML). (D) Site directed mutagenesis was performed on bonobo Schlafen11 at the indicated sites, changing them to the amino acids encoded by chimpanzee Schlafen11. Plasmids encoding wild-type or mutant Schlafen11, or chloramphenicol acetyltransferase (Chlor) as a negative control, were cotransfected into 293T cells along with a plasmid encoding GFP. Cell lysates were subject to immunoblotting. “p.s. sites” refers to positively selected sites and, in these two clones, all positions under positive selection were simultaneously changed in bonobo Schafen11 to the residues encoded in chimpanzee Schlafen11. The blots in panels A and D are representative of two independent experiments.
