@article{a2bcad3a5bea4980a0fb7fbecc46317f,
title = "Nonredundant functions of Mycobacterium tuberculosis chaperones promote survival under stress",
abstract = "Bacterial chaperones ClpB and DnaK, homologs of the respective eukaryotic heat shock proteins Hsp104 and Hsp70, are essential in the reactivation of toxic protein aggregates that occur during translation or periods of stress. In the pathogen Mycobacterium tuberculosis (Mtb), the protective effect of chaperones extends to survival in the presence of host stresses, such as protein-damaging oxidants. However, we lack a full understanding of the interplay of Hsps and other stress response genes in mycobacteria. Here, we employ genome-wide transposon mutagenesis to identify the genes that support clpB function in Mtb. In addition to validating the role of ClpB in Mtb's response to oxidants, we show that HtpG, a homolog of Hsp90, plays a distinct role from ClpB in the proteotoxic stress response. While loss of neither clpB nor htpG is lethal to the cell, loss of both through genetic depletion or small molecule inhibition impairs recovery after exposure to host-like stresses, especially reactive nitrogen species. Moreover, defects in cells lacking clpB can be complemented by overexpression of other chaperones, demonstrating that Mtb's stress response network depends upon finely tuned chaperone expression levels. These results suggest that inhibition of multiple chaperones could work in concert with host immunity to disable Mtb.",
keywords = "ClpB, Mycobacterium tuberculosis, chaperone, heat shock protein, oxidant, proteostasis",
author = "Alexa Harnagel and {Lopez Quezada}, Landys and Park, {Sae Woong} and Catherine Baranowski and Karen Kieser and Xiuju Jiang and Julia Roberts and Julien Vaubourgeix and Amy Yang and Brock Nelson and Allison Fay and Eric Rubin and Sabine Ehrt and Carl Nathan and Lupoli, {Tania J.}",
note = "Funding Information: The authors thank Kan Lin, Michael Chao, Kristin Burns‐Huang, Ben Gold, Dirk Schnappinger, Michael Glickman, Gloria Marcela Rodriguez, and Sunil Kumar for assistance with experimental interpretation and set‐up, Heran Darwin for anti‐HspR, Gang Lin for anti‐PrcB, and Jeremy Rock for CRISPRi plasmids and protocols. Rockefeller University Proteomics Resource Center (Joseph Fernandez) performed the proteomics analysis. Next‐generation sequencing was performed at the Weill Cornell Genomics Resources Core Facility (Jenny Xiang). Jamie Bean performed genomic variant analysis to identify SNPs and verify deletion strains. pEVOL‐pBpF was a gift from Peter Schultz (Addgene plasmid # 31190; http://n2t.net/addgene:31190 ; RRID:Addgene_31190). pET His6 Sumo TEV LIC cloning vector (2S‐T) was a gift from Scott Gradia (Addgene plasmid # 29711 http://n2t.net/addgene:29711 ; RRID:Addgene_29711). pHYRS52 was a gift from Hideo Iwai (Addgene plasmid # 31122; http://n2t.net/addgene:31122 ; RRID:Addgene_31122). pACYC‐T7 was a gift from Dan Bolon (Addgene plasmid # 41187; http://n2t.net/addgene:41187 ; RRID:Addgene_41187). This work was supported by NIH grant U19 AI111143 (CN, PI); Milstein Program in Chemical Biology and Translational Medicine (CN, PI); Bill & Melinda Gates Foundation (EJR, PI). K.J.K. was partially supported by a National Science Foundation Graduate Research Fellowship (Grants DGE1144152 and DGE0946799). The Department of Microbiology and Immunology at WCM is supported by the William Randolph Hearst Trust. TL acknowledges the Helen Hay Whitney and Simons Foundation, as well as NYU FAS for additional support. Funding Information: The authors thank Kan Lin, Michael Chao, Kristin Burns-Huang, Ben Gold, Dirk Schnappinger, Michael Glickman, Gloria Marcela Rodriguez, and Sunil Kumar for assistance with experimental interpretation and set-up, Heran Darwin for anti-HspR, Gang Lin for anti-PrcB, and Jeremy Rock for CRISPRi plasmids and protocols. Rockefeller University Proteomics Resource Center (Joseph Fernandez) performed the proteomics analysis. Next-generation sequencing was performed at the Weill Cornell Genomics Resources Core Facility (Jenny Xiang). Jamie Bean performed genomic variant analysis to identify SNPs and verify deletion strains. pEVOL-pBpF was a gift from Peter Schultz (Addgene plasmid # 31190; http://n2t.net/addgene:31190; RRID:Addgene_31190). pET His6 Sumo TEV LIC cloning vector (2S-T) was a gift from Scott Gradia (Addgene plasmid # 29711 http://n2t.net/addgene:29711; RRID:Addgene_29711). pHYRS52 was a gift from Hideo Iwai (Addgene plasmid # 31122; http://n2t.net/addgene:31122; RRID:Addgene_31122). pACYC-T7 was a gift from Dan Bolon (Addgene plasmid # 41187; http://n2t.net/addgene:41187; RRID:Addgene_41187). This work was supported by NIH grant U19 AI111143 (CN, PI); Milstein Program in Chemical Biology and Translational Medicine (CN, PI); Bill & Melinda Gates Foundation (EJR, PI). K.J.K. was partially supported by a National Science Foundation Graduate Research Fellowship (Grants DGE1144152 and DGE0946799). The Department of Microbiology and Immunology at WCM is supported by the William Randolph Hearst Trust. TL acknowledges the Helen Hay Whitney and Simons Foundation, as well as NYU FAS for additional support. Publisher Copyright: {\textcopyright} 2020 John Wiley & Sons Ltd",
year = "2021",
month = feb,
doi = "10.1111/mmi.14615",
language = "English (US)",
volume = "115",
pages = "272--289",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "2",
}