TY - JOUR
T1 - Analysis of small molecule ligands targeting the HIV-1 matrix protein-RNA binding site
AU - Alfadhli, Ayna
AU - McNett, Henry
AU - Eccles, Jacob
AU - Tsagli, Seyram
AU - Noviello, Colleen
AU - Sloan, Rachel
AU - López, Claudia S.
AU - Peyton, David H.
AU - Barklis, Eric
PY - 2013/1/4
Y1 - 2013/1/4
N2 - The matrix domain (MA) of the HIV-1 precursor Gag (PrGag) protein directs PrGag proteins to assembly sites at the plasma membrane by virtue of its affinity to the phospholipid, phos-phatidylinositol-4,5-bisphosphate (PI(4,5)P2). MA also binds to RNA at a site that overlaps its PI(4,5)P2 site, suggesting that RNA binding may protect MA from associating with inappropriate cellular membranes prior to PrGag delivery to the PM. Based on this, we have developed an assay in which small molecule competitors to MA-RNA binding can be characterized, with the assumption that such compounds might interfere with essential MA functions and help elucidate additional features of MA binding. Following this approach, we have identified four compounds, including three thiadiazolanes, that compete with RNA for MA binding. We also have identified MA residues involved in thiadiazolane binding and found that they overlap the MA PI(4,5)P2 and RNA sites. Cell culture studies demonstrated that thiadiazolanes inhibit HIV-1 replication but are associated with significant levels of toxicity. Nevertheless, these observations provide new insights into MA binding and pave the way for the development of antivirals that target the HIV-1 matrix domain.
AB - The matrix domain (MA) of the HIV-1 precursor Gag (PrGag) protein directs PrGag proteins to assembly sites at the plasma membrane by virtue of its affinity to the phospholipid, phos-phatidylinositol-4,5-bisphosphate (PI(4,5)P2). MA also binds to RNA at a site that overlaps its PI(4,5)P2 site, suggesting that RNA binding may protect MA from associating with inappropriate cellular membranes prior to PrGag delivery to the PM. Based on this, we have developed an assay in which small molecule competitors to MA-RNA binding can be characterized, with the assumption that such compounds might interfere with essential MA functions and help elucidate additional features of MA binding. Following this approach, we have identified four compounds, including three thiadiazolanes, that compete with RNA for MA binding. We also have identified MA residues involved in thiadiazolane binding and found that they overlap the MA PI(4,5)P2 and RNA sites. Cell culture studies demonstrated that thiadiazolanes inhibit HIV-1 replication but are associated with significant levels of toxicity. Nevertheless, these observations provide new insights into MA binding and pave the way for the development of antivirals that target the HIV-1 matrix domain.
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U2 - 10.1074/jbc.M112.399865
DO - 10.1074/jbc.M112.399865
M3 - Article
C2 - 23135280
AN - SCOPUS:84872078381
SN - 0021-9258
VL - 288
SP - 666
EP - 676
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 1
ER -