Activities of adenovirus virus-associated RNAs: purification and characterization of RNA binding proteins.
Most human adenoviruses encode two virus-associated (VA) RNAs, VA RNAI and VA RNAII, that accumulate to high levels in the cytoplasm of infected cells. The function of VA RNAI in blocking the activation of the cellular kinase PKR is well known, but the role of VA RNAII is obscure. Herein we characterize and purify several human proteins that interact preferentially with VA RNAII in Northwestern blot assays. Two of these proteins were identified as RNA helicase A and NF90, a component of the heterodimeric nuclear factor of activated T cells (NFAT). They copurified with the smaller NFAT subunit, NF45, which did not bind VA RNAII, and with an unidentified protein, p97, which did bind VA RNAII. Both RNA helicase A and NF90 contain two copies of a double-stranded (ds) RNA binding motif and bind strongly to dsRNA. NF90 interacts with RNAs in the following order of affinity: dsRNA > VA RNAII > VA RNAI > single-stranded RNA. Furthermore, VA RNAII is more effective than VA RNAI as an inhibitor of RNA helicase activity. These data identify RNA helicase A and NF90 as cellular proteins with an affinity for dsRNA and other structured RNA molecules and suggest that their functions are subject to regulation by RNA ligands including VA RNAII.
Cold Spring Harbor Laboratory, P. O. Box 100, Cold Spring Harbor, NY 11724, USA.
H J
HJ
Liao
R
R
Kobayashi
M B
MB
Mathews
D000256:Adenoviridae / Q000235:genetics*; D000595:Amino Acid Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D050778:NFATC Transcription Factors; D051903:Nuclear Factor 45 Protein; D050779:Nuclear Factor 90 Proteins; D009687:Nuclear Proteins; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D004268:DNA-Binding Proteins; C494013:ILF2 protein, human; D050778:NFATC Transcription Factors; D051903:Nuclear Factor 45 Protein; D050779:Nuclear Factor 90 Proteins; D009687:Nuclear Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014157:Transcription Factors; D012316:RNA Nucleotidyltransferases; D020365:RNA Helicases