Binding of herpes simplex virus-1 US11 to specific RNA sequences

• Published in: Nucleic acids research Vol. 33; no. 19; pp. 6090 - 6100
• Main Authors: Bryant, Kevin F., Cox, J. Colin, Wang, Hongming, Hogle, James M., Ellington, Andrew D., Coen, Donald M.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-01-2005; Oxford Publishing Limited (England)
• Subjects: Base Sequence; Binding Sites; Consensus Sequence; Electrophoretic Mobility Shift Assay; Herpesvirus 1, Human; Hydroxyl Radical - chemistry; Molecular Sequence Data; Oligonucleotides - chemistry; Protein Structure, Tertiary; Ribonucleases - metabolism; RNA - chemistry; RNA - metabolism; RNA, Double-Stranded - chemistry; RNA, Double-Stranded - metabolism; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - metabolism; Viral Proteins - chemistry; Viral Proteins - metabolism
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gki919

Herpes 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 >1014 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.