Journal
JOURNAL OF MOLECULAR BIOLOGY
Volume 433, Issue 10, Pages -Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2021.166923
Keywords
Retroviruses; Gag-RNA interactions; Purines; Footprinting; hSHAPE
Categories
Funding
- United Arab Emirates University (UAEU) Program for Advanced Research-UPAR [UPAR-31M233]
- CNRS
- UAE University Zayed Bin Sultan Center for Health Sciences [UCBR-31R123]
- [UPAR-31 M233]
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In this study, retroviral Gag proteins were found to select gRNA against spliced viral RNA by simultaneously binding to two single stranded loops on the MPMV Psi RNA. The second Gag binding site, crucial for MPMV gRNA packaging, is located immediately downstream of the major splice donor.
How retroviral Gag proteins recognize the packaging signals (Psi) on their genomic RNA (gRNA) is a key question that we addressed here using Mason-Pfizer monkey virus (MPMV) as a model system by combining band-shift assays and footprinting experiments. Our data show that Pr78(Gag) selects gRNA against spliced viral RNA by simultaneously binding to two single stranded loops on the MPMV Psi RNA: (1) a large purine loop (ssPurines), and (2) a loop which partially overlaps with a mostly base-paired purine repeat (bpPurines) and extends into a GU-rich binding motif. Importantly, this second Gag binding site is located immediately downstream of the major splice donor (mSD) and is thus absent from the spliced viral RNAs. Identifying elements crucial for MPMV gRNA packaging should help in understanding not only the mechanism of virion assembly by retroviruses, but also facilitate construction of safer retroviral vectors for human gene therapy. (C) 2021 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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