Essential host cofactors in retrovirus replication bind cis-acting sequences in the 5'untranslated region (UTR). Although host RBPs are crucial to all aspects of virus biology, elucidating their roles in replication remains a challenge to the field. Here RNA affinity-coupled-proteomics generated a comprehensive, unbiased inventory of human and avian RNA binding proteins (RBPs) co-isolating with 5'UTRs of HIV-1, spleen necrosis virus and Rous sarcoma virus. Applying stringent biochemical and statistical criteria, we identified 185 RBP; 122 were previously implicated in retrovirus biology and 63 are new to the 5'UTR proteome. RNA electrophoretic mobility assays investigated paralogs present in the common ancestor of vertebrates and one hnRNP was identified as a central node to the biological process-anchored networks of HIV-1, SNV, and RSV 5' UTR-proteomes. This comprehensive view of the host constituents of retroviral RNPs is broadly applicable to investigation of viral replication and antiviral response in both human and avian cell lineages.
Bibliographical noteFunding Information:
We thank Bruce Stanley, PhD, Penn State College of Medicine Proteomics and Small Molecule Mass Spectrometry Core Facility, for help with data analysis using Protein Pilot. This research was performed with support by the National Institutes of Health’s National Institutes of General Medical Sciences P50GM103297 to LJP and KBL ; and National Cancer Institute P01CA16058 to The Ohio State University Comprehensive Cancer Center and support from NIH National Cancer Institute to LJP (R01 CA76534) and RJK (F31 CA171862) .
© 2015 Elsevier Inc.
- Cis-Acting replication sequences
- Post-transcriptional control
- RNA recognition motif
- Virus-host interaction