TY - JOUR
T1 - SdhA blocks disruption of the Legionella-containing vacuole by hijacking the OCRL phosphatase
AU - Choi, Won Young
AU - Kim, Seongok
AU - Aurass, Philipp
AU - Huo, Wenwen
AU - Creasey, Elizabeth A
AU - Edwards, Marc
AU - Lowe, Martin
AU - Isberg, Ralph R
N1 - Funding Information:
This work was supported by HHMI and NIAID grants R01 AI113211 and R01 AI146245 to R.R.I. P.A. was funded by DFG grant AU550/1-1 . M.E. is supported by NIH grant 1R15GM143733 from NIGMS . We thank Ila Anand for scientific discussions throughout the course of this work and Kristen Davis and Erion Lipo for review of the manuscript. We thank Dr. Pietro De Camilli for providing OCRL antibody and GFP-OCRL construct and Dr. Matthias Machner for Rab5a (Q79L) construct. We thank Ross Tomaino of Taplin Mass Spectrometry Facility at Harvard Medical School for performing MS/MS analysis as well as for numerous consultations regarding data analysis. We also thank Drs. Elizabeth Draganova and Ellen White for patient help setting up the lipid extrusion assays and with baculovirus expression.
Funding Information:
This work was supported by HHMI and NIAID grants R01 AI113211 and R01 AI146245 to R.R.I. P.A. was funded by DFG grant AU550/1-1. M.E. is supported by NIH grant 1R15GM143733 from NIGMS. We thank Ila Anand for scientific discussions throughout the course of this work and Kristen Davis and Erion Lipo for review of the manuscript. We thank Dr. Pietro De Camilli for providing OCRL antibody and GFP-OCRL construct and Dr. Matthias Machner for Rab5a (Q79L) construct. We thank Ross Tomaino of Taplin Mass Spectrometry Facility at Harvard Medical School for performing MS/MS analysis as well as for numerous consultations regarding data analysis. We also thank Drs. Elizabeth Draganova and Ellen White for patient help setting up the lipid extrusion assays and with baculovirus expression. W.Y.C. and R.R.I. conceived of and designed this study. W.Y.C. and R.R.I. wrote the manuscript with input from all authors. W.Y.C. S.K. P.A. and E.A.C. performed experiments and constructed strains expressly for this work. W.H. performed bioinformatic analysis. M.E. and M.L. provided strains, reagents, plasmids, and unpublished procedures. M.E. M.L. and R.R.I. provided experimental guidance. The authors declare no competing interests.
Publisher Copyright:
© 2021 The Author(s)
PY - 2021/11/2
Y1 - 2021/11/2
N2 - Legionella pneumophila grows intracellularly within a replication vacuole via action of Icm/Dot-secreted proteins. One such protein, SdhA, maintains the integrity of the vacuolar membrane, thereby preventing cytoplasmic degradation of bacteria. We show here that SdhA binds and blocks the action of OCRL (OculoCerebroRenal syndrome of Lowe), an inositol 5-phosphatase pivotal for controlling endosomal dynamics. OCRL depletion results in enhanced vacuole integrity and intracellular growth of a sdhA mutant, consistent with OCRL participating in vacuole disruption. Overexpressed SdhA alters OCRL function, enlarging endosomes, driving endosomal accumulation of phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2), and interfering with endosomal trafficking. SdhA interrupts Rab guanosine triphosphatase (GTPase)-OCRL interactions by binding to the OCRL ASPM-SPD2-Hydin (ASH) domain, without directly altering OCRL 5-phosphatase activity. The Legionella vacuole encompassing the sdhA mutant accumulates OCRL and endosomal antigen EEA1 (Early Endosome Antigen 1), consistent with SdhA blocking accumulation of OCRL-containing endosomal vesicles. Therefore, SdhA hijacking of OCRL is associated with blocking trafficking events that disrupt the pathogen vacuole.
AB - Legionella pneumophila grows intracellularly within a replication vacuole via action of Icm/Dot-secreted proteins. One such protein, SdhA, maintains the integrity of the vacuolar membrane, thereby preventing cytoplasmic degradation of bacteria. We show here that SdhA binds and blocks the action of OCRL (OculoCerebroRenal syndrome of Lowe), an inositol 5-phosphatase pivotal for controlling endosomal dynamics. OCRL depletion results in enhanced vacuole integrity and intracellular growth of a sdhA mutant, consistent with OCRL participating in vacuole disruption. Overexpressed SdhA alters OCRL function, enlarging endosomes, driving endosomal accumulation of phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2), and interfering with endosomal trafficking. SdhA interrupts Rab guanosine triphosphatase (GTPase)-OCRL interactions by binding to the OCRL ASPM-SPD2-Hydin (ASH) domain, without directly altering OCRL 5-phosphatase activity. The Legionella vacuole encompassing the sdhA mutant accumulates OCRL and endosomal antigen EEA1 (Early Endosome Antigen 1), consistent with SdhA blocking accumulation of OCRL-containing endosomal vesicles. Therefore, SdhA hijacking of OCRL is associated with blocking trafficking events that disrupt the pathogen vacuole.
KW - Legionella pneumophila
KW - OCRL
KW - bacterial pathogenesis
KW - endosomes
KW - intracellular replication
KW - vesicle trafficking
UR - https://www.scopus.com/pages/publications/85118504110
U2 - 10.1016/j.celrep.2021.109894
DO - 10.1016/j.celrep.2021.109894
M3 - Article
C2 - 34731604
SN - 2211-1247
VL - 37
SP - 109894
JO - Cell Reports
JF - Cell Reports
IS - 5
M1 - 109894
ER -