TY - JOUR
T1 - Two Faces of CwlM, an Essential PknB Substrate, in Mycobacterium tuberculosis
AU - Turapov, Obolbek
AU - Forti, Francesca
AU - Kadhim, Baleegh
AU - Ghisotti, Daniela
AU - Sassine, Jad
AU - Straatman-Iwanowska, Anna
AU - Bottrill, Andrew R.
AU - Moynihan, Patrick J.
AU - Wallis, Russell
AU - Barthe, Philippe
AU - Cohen-Gonsaud, Martin
AU - Ajuh, Paul
AU - Vollmer, Waldemar
AU - Mukamolova, Galina V.
PY - 2018/10/2
Y1 - 2018/10/2
N2 - Tuberculosis claims >1 million lives annually, and its causative agent Mycobacterium tuberculosis is a highly successful pathogen. Protein kinase B (PknB) is reported to be critical for mycobacterial growth. Here, we demonstrate that PknB-depleted M. tuberculosis can replicate normally and can synthesize peptidoglycan in an osmoprotective medium. Comparative phosphoproteomics of PknB-producing and PknB-depleted mycobacteria identify CwlM, an essential regulator of peptidoglycan synthesis, as a major PknB substrate. Our complementation studies of a cwlM mutant of M. tuberculosis support CwlM phosphorylation as a likely molecular basis for PknB being essential for mycobacterial growth. We demonstrate that growing mycobacteria produce two forms of CwlM: a non-phosphorylated membrane-associated form and a PknB-phosphorylated cytoplasmic form. Furthermore, we show that the partner proteins for the phosphorylated and non-phosphorylated forms of CwlM are FhaA, a fork head-associated domain protein, and MurJ, a proposed lipid II flippase, respectively. From our results, we propose a model in which CwlM potentially regulates both the biosynthesis of peptidoglycan precursors and their transport across the cytoplasmic membrane. PknB controls growth and peptidoglycan biosynthesis in Mycobacterium tuberculosis. Turapov et al. show that CwlM, a major PknB substrate, is produced in two forms: a non-phosphorylated membrane-associated CwlM and a PknB-phosphorylated cytoplasmic CwlM. The phosphorylated CwlM binds FhaA, a fork head-associated domain protein, while non-phosphorylated CwlM interacts with MurJ (MviN), a proposed lipid II flippase.
AB - Tuberculosis claims >1 million lives annually, and its causative agent Mycobacterium tuberculosis is a highly successful pathogen. Protein kinase B (PknB) is reported to be critical for mycobacterial growth. Here, we demonstrate that PknB-depleted M. tuberculosis can replicate normally and can synthesize peptidoglycan in an osmoprotective medium. Comparative phosphoproteomics of PknB-producing and PknB-depleted mycobacteria identify CwlM, an essential regulator of peptidoglycan synthesis, as a major PknB substrate. Our complementation studies of a cwlM mutant of M. tuberculosis support CwlM phosphorylation as a likely molecular basis for PknB being essential for mycobacterial growth. We demonstrate that growing mycobacteria produce two forms of CwlM: a non-phosphorylated membrane-associated form and a PknB-phosphorylated cytoplasmic form. Furthermore, we show that the partner proteins for the phosphorylated and non-phosphorylated forms of CwlM are FhaA, a fork head-associated domain protein, and MurJ, a proposed lipid II flippase, respectively. From our results, we propose a model in which CwlM potentially regulates both the biosynthesis of peptidoglycan precursors and their transport across the cytoplasmic membrane. PknB controls growth and peptidoglycan biosynthesis in Mycobacterium tuberculosis. Turapov et al. show that CwlM, a major PknB substrate, is produced in two forms: a non-phosphorylated membrane-associated CwlM and a PknB-phosphorylated cytoplasmic CwlM. The phosphorylated CwlM binds FhaA, a fork head-associated domain protein, while non-phosphorylated CwlM interacts with MurJ (MviN), a proposed lipid II flippase.
KW - cellular localization
KW - CwlM
KW - MurJ
KW - Mycobacterium tuberculosis
KW - peptidoglycan
KW - phosphoproteomics
KW - protein kinase B
KW - serine/threonine protein kinase
UR - http://www.scopus.com/inward/record.url?scp=85053692176&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2018.09.004
DO - 10.1016/j.celrep.2018.09.004
M3 - Article
C2 - 30282038
AN - SCOPUS:85053692176
SN - 2211-1247
VL - 25
SP - 57-67.e5
JO - Cell Reports
JF - Cell Reports
IS - 1
ER -