title: Acyldepsipeptide antibiotics kill mycobacteria by preventing the physiological functions of the ClpP1P2 protease.
abstract: The Clp protease complex in Mycobacterium tuberculosis is unusual in its composition, functional importance and activation mechanism. Whilst most bacterial species contain a single ClpP protein that is dispensable for normal growth, mycobacteria have two ClpPs, ClpP1 and ClpP2, which are essential for viability and together form the ClpP1P2 tetradecamer. Acyldepsipeptide antibiotics of the ADEP class inhibit the growth of Gram-positive firmicutes by activating ClpP and causing unregulated protein degradation. Here we show that, in contrast, mycobacteria are killed by ADEP through inhibition of ClpP function. Although ADEPs can stimulate purified M. tuberculosis ClpP1P2 to degrade larger peptides and unstructured proteins, this effect is weaker than for ClpP from other bacteria and depends on the presence of an additional activating factor (e.g. the dipeptide benzyloxycarbonyl-leucyl-leucine in vitro) to form the active ClpP1P2 tetradecamer. The cell division protein FtsZ, which is a particularly sensitive target for ADEP-activated ClpP in firmicutes, is not degraded in mycobacteria. Depletion of the ClpP1P2 level in a conditional Mycobacterium bovis BCG mutant enhanced killing by ADEP unlike in other bacteria. In summary, ADEPs kill mycobacteria by preventing interaction of ClpP1P2 with the regulatory ATPases, ClpX or ClpC1, thus inhibiting essential ATP-dependent protein degradation.
authors.affiliation: Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tuebingen, Tuebingen, Germany.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Famulla
authors.affiliation: Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tuebingen, Tuebingen, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Sass
authors.affiliation: Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tuebingen, Tuebingen, Germany.
authors.forename: Imran
authors.initials: I
authors.lastname: Malik
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
authors.forename: Tatos
authors.initials: T
authors.lastname: Akopian
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Kandror
authors.affiliation: Institute for Medical Microbiology and Hospital Hygiene, University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Marina
authors.initials: M
authors.lastname: Alber
authors.affiliation: Bayer Pharma AG, Wuppertal, Germany.
authors.forename: Berthold
authors.initials: B
authors.lastname: Hinzen
authors.affiliation: AiCuris GmbH & Co. KG, Wuppertal, Germany.
authors.forename: Helga
authors.initials: H
authors.lastname: Ruebsamen-Schaeff
authors.affiliation: Institute for Medical Microbiology and Hospital Hygiene, University of Duesseldorf, Duesseldorf, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Kalscheuer
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
authors.forename: Alfred L
authors.initials: AL
authors.lastname: Goldberg
authors.affiliation: Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tuebingen, Tuebingen, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Brötz-Oesterhelt
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000187:drug effects / Q000378:metabolism; D047630:Depsipeptides / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D049071:Endopeptidase Clp / Q000187:drug effects* / Q000378:metabolism* / Q000502:physiology; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D059748:Proteolysis; D012697:Serine Endopeptidases / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D047630:Depsipeptides; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D049071:Endopeptidase Clp; D000251:Adenosine Triphosphatases