Supplemental figure legends9 Figure S1. Related to Figure 1. Cell and nucleoid morphology of E. coli cells in different growth 10 media. 11 A. Representative phase contrast and DAPI images of E. coli cells (CJW6324) grown in liquid cultures 12of M9 medium supplemented with the indicated carbon source and other chemicals (CAAT: 0.1% 13 casamino acids and 1 µg/ml thiamine) at 37 °C. For a full description of the growth media, see Table 14 S1. Cell contours (green) were generated using Oufti. 15 B. Bar graph showing the average doubling times of cultures when growing in exponential phase in 16 the indicated growth media. Errors bars indicate the standard deviation between three independent 17 biological replicates. Colors correspond to those used in Figure 1B. 18 C. Scatter plot of growth medium osmolality versus average NC ratio for E. coli cells (CJW6324) grown 19 in the media indicated in B. The color scheme corresponds to the one shown in B. Error bars indicate 20 95% confidence intervals. 21 22
Using a computationally driven approach, a class of inhibitors with picomolar potency known as the catechol diethers were developed targeting the non-nucleoside binding pocket (NNBP) of HIV-1 RT. Computational studies suggested that halogen bonding interactions between the C5 substituent of the inhibitor and backbone carbonyl of conserved residue Pro95 might be important. While the recently reported crystal structures of the RT complexes confirmed the interactions with the NNBP, they revealed the lack of a halogen bonding interaction with Pro95. In order to understand the effects of substituents at the C5 position, we determined additional crystal structures with 5-Br and 5-H derivatives. Using comparative structural analysis, we identified several conformations of the ethoxy uracil dependent on the strength of a van der Waals interaction with the Cγ of Pro95 and the C5 substitution. The 5-Cl and 5-F derivatives position the ethoxy uracil to make more hydrogen bonds, while the larger 5-Br and smaller 5-H position the ethoxy uracil to make fewer hydrogen bonds. EC50 values correlate with the trends observed in the crystal structures. The influence of C5 substitutions on the ethoxy uracil conformation may have strategic value, as future derivatives can possibly be modulated in order to gain additional hydrogen bonding interactions with resistant variants of RT.
Catechol diether compounds have nanomolar antiviral and enzymatic activity against HIV with reverse transcriptase (RT) variants containing K101P, a mutation that confers high-level resistance to FDA-approved non-nucleoside inhibitors efavirenz and rilpivirine. Kinetic data suggests that RT (K101P) variants are as catalytically fit as wild-type and thus can potentially increase in the viral population as more antiviral regimens include efavirenz or rilpivirine. Comparison of wild-type structures and a new crystal structure of RT (K101P) in complex with a leading compound confirms that the K101P mutation is not a liability for the catechol diethers while suggesting that key interactions are lost with efavirenz and rilpivirine.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.