Binding of the polypyrimidine tract-binding protein-associated splicing factor (PSF) to the hepatitis delta virus RNA

• Published in: Virology (New York, N.Y.) Vol. 356; no. 1; pp. 35 - 44
• Main Authors: Greco-Stewart, Valerie S., Thibault, Catherine St-Laurent, Pelchat, Martin
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2006
• Subjects: 60 APPLIED LIFE SCIENCES; Amino Acid Sequence; Base Sequence; Co-immunoprecipitation; CROSS-LINKING; Cross-Linking Reagents; HELA CELLS; HEPATITIS; Hepatitis D virus; Hepatitis delta virus; Hepatitis Delta Virus - genetics; Hepatitis Delta Virus - metabolism; Humans; Immunoprecipitation; IN VITRO; IN VIVO; LIFE CYCLE; Mass Spectrometry; MASS SPECTROSCOPY; Molecular Sequence Data; Polypyrimidine tract-binding protein; Polypyrimidine Tract-Binding Protein - metabolism; PROTEINS; PSF; PTB-Associated Splicing Factor; RNA; RNA binding protein; RNA replication; RNA Splicing; RNA virus; RNA, Viral - genetics; RNA, Viral - metabolism; RNA-Binding Proteins - metabolism; SPECIFICITY; SPLICING; Splicing factor; Ultraviolet Rays; UV cross-linking; VIRUSES; UV cross-linking; RNA replication; PSF; Co-immunoprecipitation; RNA virus; RNA binding protein; Splicing factor; Mass spectrometry; Hepatitis delta virus; Polypyrimidine tract-binding protein
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/j.virol.2006.06.040

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.