Binding of the polypyrimidine tract-binding protein-associated splicing factor (PSF) to the hepatitis delta virus RNA.
The hepatitis delta virus (HDV) has a very limited protein coding capacity and must rely on host proteins for its replication. A ribonucleoprotein complex was detected following UV cross-linking between HeLa nuclear proteins and an RNA corresponding to the right terminal stem-loop domain of HDV genomic RNA. Mass spectrometric analysis of the complex revealed the polypyrimidine tract-binding protein-associated splicing factor (PSF) as a novel HDV RNA-interacting protein. Co-immunoprecipitation demonstrated the interaction between HDV RNA and PSF both in vitro in HeLa nuclear extract and in vivo within HeLa cells containing both polarities of the HDV genome. Analysis of the binding of various HDV-derived RNAs to purified, recombinant PSF further confirmed the specificity of the interaction and revealed that PSF directly binds to the terminal stem-loop domains of both polarities of HDV RNA. Our findings provide evidence of the involvement of a host mRNA processing protein in the HDV life cycle.
Greco-Stewart
Valerie S
VS
Department of Biochemistry, Faculty of Medicine, University of Ottawa, 451 Smyth Road, Room 4223A, Ottawa, ON, Canada K1H 8M5.
Thibault
Catherine St-Laurent
CS
Pelchat
Martin
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D003432:Cross-Linking Reagents; D006367:HeLa Cells; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D000072670:PTB-Associated Splicing Factor; D038941:Polypyrimidine Tract-Binding Protein / Q000378:metabolism; D012326:RNA Splicing; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D000072670:PTB-Associated Splicing Factor; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D038941:Polypyrimidine Tract-Binding Protein