Bryant, Kevin F.Cox, J. ColinWang, HongmingHogle, James M.Ellington, Andrew D.Coen, Donald M.2019-10-052005Bryant, K. F. 2005. “Binding of Herpes Simplex Virus-1 US11 to Specific RNA Sequences.” Nucleic Acids Research 33 (19): 6090–6100. https://doi.org/10.1093/nar/gki919.0305-10481362-4962http://nrs.harvard.edu/urn-3:HUL.InstRepos:41482943Herpes simplex virus-1 US11 is a RNA-binding protein with a novel RNA-binding domain. US11 has been reported to exhibit sequence- and conformation-specific RNA-binding, but the sequences and conformations important for binding are not known. US11 has also been described as a double-stranded RNA (dsRNA)-binding protein. To investigate the US11-RNA interaction, we performed in vitro selection of RNA aptamers that bind US11 from a RNA library consisting of > 10(14) 80 base sequences which differ in a 30 base randomized region. US11 bound specifically to selected aptamers with an affinity of 70 nM. Analysis of 23 selected sequences revealed a strong consensus sequence. The US11 RNA-binding domain and <= 46 bases of selected RNA containing the consensus sequence were each sufficient for binding. US11 binding protected the consensus motif from hydroxyl radical cleavage. RNase digestions of a selected aptamer revealed regions of both single-stranded RNA and dsRNA. We observed that US11 bound two different dsRNAs in a sequence non-specific manner, but with lower affinity than it bound selected aptamers. The results define a relatively short specific sequence that binds US11 with high affinity and indicate that dsRNA alone does not confer high-affinity binding.en-USJournal Article2019-10-0510.1093/nar/gki919