TY - JOUR
T1 - Structural mechanism of transcription inhibition by lasso peptides microcin J25 and capistruin
AU - Braffman, Nathaniel R.
AU - Piscotta, Frank J.
AU - Hauver, Jesse
AU - Campbell, Elizabeth A.
AU - James Link, A.
AU - Darst, Seth A.
N1 - Funding Information:
We thank B. Bae, K.-A. Twist, M. Chaker-Margot, P. Olinares, and B. T. Chait for initial contributions to this project. This work is based on research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by National Institute of General Medical Sciences–National Institutes of Health (NIH) Grant P41 GM103403.The Pila-tus 6M detector on 24-ID-C beamline is funded by NIH-Office of Research Infrastructure Programs High-End Instruments Grant S10 RR029205. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. The use of The Rockefeller University Structural Biology Resource Center was made possible by NIH/National Center for Research Resources Grant 1S10RR027037. This work was supported by NIH Grants R01GM107036(to A.J.L.) and R35 GM118130 (to S.A.D.).
Funding Information:
ACKNOWLEDGMENTS. We thank B. Bae, K.-A. Twist, M. Chaker-Margot, P. Olinares, and B. T. Chait for initial contributions to this project. This work is based on research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by National Institute of General Medical Sciences–National Institutes of Health (NIH) Grant P41 GM103403. The Pila-tus 6M detector on 24-ID-C beamline is funded by NIH-Office of Research Infrastructure Programs High-End Instruments Grant S10 RR029205. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under ContractDE-AC02-06CH11357. The use of The Rockefeller University Structural Biology Resource Center was made possible by NIH/National Center for Research Resources Grant 1S10RR027037. This work was supported by NIH Grants R01 GM107036 (to A.J.L.) and R35 GM118130 (to S.A.D.).
Publisher Copyright:
© 2019 National Academy of Sciences. All Rights Reserved.
PY - 2019/1/22
Y1 - 2019/1/22
N2 - We report crystal structures of the antibacterial lasso peptides microcin J25 (MccJ25) and capistruin (Cap) bound to their natural enzymatic target, the bacterial RNA polymerase (RNAP). Both peptides bind within the RNAP secondary channel, through which NTP substrates enter the RNAP active site, and sterically block trigger-loop folding, which is essential for efficient catalysis by the RNAP. MccJ25 binds deep within the secondary channel in a manner expected to interfere with NTP substrate binding, explaining the partial competitive mechanism of inhibition with respect to NTPs found previously [Mukhopadhyay J, Sineva E, Knight J, Levy RM, Ebright RH (2004) Mol Cell 14:739–751]. The Cap binding determinant on RNAP overlaps, but is not identical to, that of MccJ25. Cap binds further from the RNAP active site and does not sterically interfere with NTP binding, and we show that Cap inhibition is partially noncompetitive with respect to NTPs. This work lays the groundwork for structure determination of other lasso peptides that target the bacterial RNAP and provides a structural foundation to guide lasso peptide antimicrobial engineering approaches.
AB - We report crystal structures of the antibacterial lasso peptides microcin J25 (MccJ25) and capistruin (Cap) bound to their natural enzymatic target, the bacterial RNA polymerase (RNAP). Both peptides bind within the RNAP secondary channel, through which NTP substrates enter the RNAP active site, and sterically block trigger-loop folding, which is essential for efficient catalysis by the RNAP. MccJ25 binds deep within the secondary channel in a manner expected to interfere with NTP substrate binding, explaining the partial competitive mechanism of inhibition with respect to NTPs found previously [Mukhopadhyay J, Sineva E, Knight J, Levy RM, Ebright RH (2004) Mol Cell 14:739–751]. The Cap binding determinant on RNAP overlaps, but is not identical to, that of MccJ25. Cap binds further from the RNAP active site and does not sterically interfere with NTP binding, and we show that Cap inhibition is partially noncompetitive with respect to NTPs. This work lays the groundwork for structure determination of other lasso peptides that target the bacterial RNAP and provides a structural foundation to guide lasso peptide antimicrobial engineering approaches.
KW - Capistruin
KW - Lasso peptide
KW - Microcin J25
KW - RNA polymerase
KW - X-ray crystallography
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U2 - 10.1073/pnas.1817352116
DO - 10.1073/pnas.1817352116
M3 - Article
C2 - 30626643
AN - SCOPUS:85060314869
SN - 0027-8424
VL - 116
SP - 1273
EP - 1278
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 4
ER -