title: Chlamydia trachomatis co-opts GBF1 and CERT to acquire host sphingomyelin for distinct roles during intracellular development.
abstract: The strain designated Chlamydia trachomatis serovar that was used for experiments in this paper is Chlamydia muridarum, a species closely related to C. trachomatis (and formerly termed the Mouse Pneumonitis strain of C. trachomatis. [corrected]. The obligate intracellular pathogen Chlamydia trachomatis replicates within a membrane-bound inclusion that acquires host sphingomyelin (SM), a process that is essential for replication as well as inclusion biogenesis. Previous studies demonstrate that SM is acquired by a Brefeldin A (BFA)-sensitive vesicular trafficking pathway, although paradoxically, this pathway is dispensable for bacterial replication. This finding suggests that other lipid transport mechanisms are involved in the acquisition of host SM. In this work, we interrogated the role of specific components of BFA-sensitive and BFA-insensitive lipid trafficking pathways to define their contribution in SM acquisition during infection. We found that C. trachomatis hijacks components of both vesicular and non-vesicular lipid trafficking pathways for SM acquisition but that the SM obtained from these separate pathways is being utilized by the pathogen in different ways. We show that C. trachomatis selectively co-opts only one of the three known BFA targets, GBF1, a regulator of Arf1-dependent vesicular trafficking within the early secretory pathway for vesicle-mediated SM acquisition. The Arf1/GBF1-dependent pathway of SM acquisition is essential for inclusion membrane growth and stability but is not required for bacterial replication. In contrast, we show that C. trachomatis co-opts CERT, a lipid transfer protein that is a key component in non-vesicular ER to trans-Golgi trafficking of ceramide (the precursor for SM), for C. trachomatis replication. We demonstrate that C. trachomatis recruits CERT, its ER binding partner, VAP-A, and SM synthases, SMS1 and SMS2, to the inclusion and propose that these proteins establish an on-site SM biosynthetic factory at or near the inclusion. We hypothesize that SM acquired by CERT-dependent transport of ceramide and subsequent conversion to SM is necessary for C. trachomatis replication whereas SM acquired by the GBF1-dependent pathway is essential for inclusion growth and stability. Our results reveal a novel mechanism by which an intracellular pathogen redirects SM biosynthesis to its replicative niche.
authors: Department of Medicine, University of California, San Francisco, California, United States of America.
authors: Cherilyn A
authors: CA
authors: Elwell
authors: Shaobo
authors: S
authors: Jiang
authors: Jung Hwa
authors: JH
authors: Kim
authors: Albert
authors: A
authors: Lee
authors: Torsten
authors: T
authors: Wittmann
authors: Kentaro
authors: K
authors: Hanada
authors: Paul
authors: P
authors: Melancon
authors: Joanne N
authors: JN
authors: Engel
mesh_terms: D000577:Amides / Q000494:pharmacology; D001549:Benzamides / Q000494:pharmacology; D001565:Benzoates / Q000494:pharmacology; D020126:Brefeldin A / Q000494:pharmacology; D047389:Casein Kinase I / Q000378:metabolism; D002692:Chlamydia trachomatis / Q000254:growth & development / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D002479:Inclusion Bodies / Q000502:physiology*; D008565:Membrane Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D013109:Sphingomyelins / Q000096:biosynthesis*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism*
chemical_list: C568977:2-(4-fluorobenzoylamino)benzoic acid methyl ester; C119087:ARFGEF1 protein, human; C119088:ARFGEF2 protein, human; D000577:Amides; D001549:Benzamides; D001565:Benzoates; C120445:GBF1 protein, human; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; C441643:N-(3-hydroxy-1-hydroxymethyl-3-phenylpropyl)dodecanamide; D009419:Nerve Tissue Proteins; D013109:Sphingomyelins; C114051:VAPA protein, human; D033921:Vesicular Transport Proteins; D020126:Brefeldin A; C119090:CERT1 protein, human; D047389:Casein Kinase I; D017346:Protein Serine-Threonine Kinases; C455440:SGMS1 protein, human; D017855:Transferases (Other Substituted Phosphate Groups)