Ribonuclease zymogen induces cytotoxicity upon HIV-1 infection
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12981_2021_Article_399.pdf
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Author(s)
Windsor, Ian W.
Dudley, Dawn M.
O’Connor, David H.
Raines, Ronald T.
Date Issued
October 26, 2021
Publisher
BioMed Central
Citation
AIDS Research and Therapy. 2021 Oct 26;18(1):77
Version
Final published version
Abstract
Abstract
Background
Targeting RNA is a promising yet underdeveloped modality for the selective killing of cells infected with HIV-1. The secretory ribonucleases (RNases) found in vertebrates have cytotoxic ribonucleolytic activity that is kept in check by a cytosolic ribonuclease inhibitor protein, RI.
Methods
We engineered amino acid substitutions that enable human RNase 1 to evade RI upon its cyclization into a zymogen that is activated by the HIV-1 protease. In effect, the zymogen has an HIV-1 protease cleavage site between the termini of the wild-type enzyme, thereby positioning a cleavable linker over the active site that blocks access to a substrate.
Results
The amino acid substitutions in RNase 1 diminish its affinity for RI by 106-fold and confer high toxicity for T-cell leukemia cells. Pretreating these cells with the zymogen leads to a substantial drop in their viability upon HIV-1 infection, indicating specific toxicity toward infected cells.
Conclusions
These data demonstrate the utility of ribonuclease zymogens as biologic prodrugs.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Koch Institute for Integrative Cancer Research at MIT
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1186/s12981-021-00399-z
