Tuberculosis kills 1.5-2 million people globally every year. An effective vaccine or chemotherapy has yet to be developed. Recently, through a large-scale transposon mutagenesis screening, the Mycobacterium tuberculosis (Mtb) proteasome and the proteasomal ATPase (Mpa) were found to be required for Mtb resistance to killing by a source of nitric oxide (NO). NO is produced by the host immune system to control Mtb infections. Proteasome and Mpa appear to protect Mtb against NO by degrading proteins after exposure to NO. Thus, Mpa and the Mtb proteasome may be promising targets for the development of anti-Tb chemotherapeutics. Mtb proteasome has unique broad substrate specificity towards small synthetic peptides, and its proteolytic activity is inhibited by MLN-273, an analog of the anti-myeloma drug bortezomib that targets human proteasome. The unique substrate binding property of Mtb proteasome can be exploited to develop Mtb specific inhibitors. Mpa is homologous to ATPases that function with the eukaryotic proteasome, and likely unfolds the protein substrate for processive degradation by Mtb proteasome. A mutant Mpa protein missing only its last two amino acids retains ATPase activity, yet fails to protect Mtb against nitrite. We propose to use a combination of cryo-electron microscopy (cryo-EM) and X-ray crystallography to investigate the structures of Mtb proteasome and Mpa. Our comprehensive structural studies (1) will reveal the structural basis of the Mtb proteasome's broad substrate specificity; (2) will uncover the inhibition mechanism of the dipeptidyl boronate MLN-273; (3) will elucidate the conformational changes of Mpa associated with the ATP hydrolysis cycle; (4) will provide answer to the long standing question as to how a symmetry mismatched six-fold ATPase oligomer activates the seven-fold proteasome, (5) will contribute to a better understanding of the eubacterial proteasome biology, and (6) will set the stage for the structure-based anti-TB chemotherapeutic development. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI070285-01A1
Application #
7259003
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Sizemore, Christine F
Project Start
2007-04-01
Project End
2012-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
1
Fiscal Year
2007
Total Cost
$369,891
Indirect Cost
Name
Brookhaven National Laboratory
Department
Type
DUNS #
027579460
City
Upton
State
NY
Country
United States
Zip Code
11973
Hu, Kuan; Jastrab, Jordan B; Zhang, Susan et al. (2018) Proteasome substrate capture and gate opening by the accessory factor PafE from Mycobacterium tuberculosis. J Biol Chem 293:4713-4723
Yu, Hongjun; Lupoli, Tania J; Kovach, Amanda et al. (2018) ATP hydrolysis-coupled peptide translocation mechanism of Mycobacterium tuberculosis ClpB. Proc Natl Acad Sci U S A 115:E9560-E9569
Samanovic, Marie I; Hsu, Hao-Chi; Jones, Marcus B et al. (2018) Cytokinin Signaling in Mycobacterium tuberculosis. MBio 9:
Yu, Hongjun; Wu, Chang-Hao; Schut, Gerrit J et al. (2018) Structure of an Ancient Respiratory System. Cell 173:1636-1649.e16
Zhang, Susan; Burns-Huang, Kristin E; Janssen, Guido V et al. (2017) Mycobacterium tuberculosis Proteasome Accessory Factor A (PafA) Can Transfer Prokaryotic Ubiquitin-Like Protein (Pup) between Substrates. MBio 8:
Wu, Yujie; Hu, Kuan; Li, Defeng et al. (2017) Mycobacterium tuberculosis proteasomal ATPase Mpa has a ?-grasp domain that hinders docking with the proteasome core protease. Mol Microbiol 105:227-241
Bai, Lin; Jastrab, Jordan B; Isasa, Marta et al. (2017) Structural Analysis of Mycobacterium tuberculosis Homologues of the Eukaryotic Proteasome Assembly Chaperone 2 (PAC2). J Bacteriol 199:
Hsu, Hao-Chi; Singh, Pradeep K; Fan, Hao et al. (2017) Structural Basis for the Species-Selective Binding of N,C-Capped Dipeptides to the Mycobacterium tuberculosis Proteasome. Biochemistry 56:324-333
Tian, Ye; Zhang, Yugang; Wang, Tong et al. (2016) Lattice engineering through nanoparticle-DNA frameworks. Nat Mater 15:654-61
Liu, Wenyan; Tagawa, Miho; Xin, Huolin L et al. (2016) Diamond family of nanoparticle superlattices. Science 351:582-6

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