title: Beclin 1 is required for starvation-enhanced, but not rapamycin-enhanced, LC3-associated phagocytosis of Burkholderia pseudomallei in RAW 264.7 cells.
abstract: LC3-associated phagocytosis (LAP) of Burkholderia pseudomallei by murine macrophage (RAW 264.7) cells is an intracellular innate defense mechanism. Beclin 1, a protein with several roles in autophagic processes, is known to be recruited to phagosomal membranes as a very early event in LAP. We sought to determine whether knockdown of Beclin 1 by small interfering RNA (siRNA) would affect recruitment of LC3 and subsequent LAP of infecting B. pseudomallei. Both starvation and rapamycin treatment can induce Beclin 1-dependent autophagy. Therefore, we analyzed the consequences of Beclin 1 knockdown for LAP in infected cells that had been either starved or treated with rapamycin by determining the levels of bacterial colocalization with LC3 and intracellular survival. Concurrently, we confirmed the location of bacteria as either contained in phagosomes or free in the cytoplasm. We found that both rapamycin and starvation treatment enhanced LAP of B. pseudomallei but that the rapamycin response is Beclin 1 independent whereas the starvation response is Beclin 1 dependent.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton campus, Victoria, Australia.
authors.forename: Xuelei
authors.initials: X
authors.lastname: Li
authors.forename: Mark
authors.initials: M
authors.lastname: Prescott
authors.forename: Ben
authors.initials: B
authors.lastname: Adler
authors.forename: John D
authors.initials: JD
authors.lastname: Boyce
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Devenish
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001343:Autophagy / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D016957:Burkholderia pseudomallei / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D008554:Melioidosis / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010587:Phagocytosis / Q000235:genetics / Q000276:immunology*; D010588:Phagosomes / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology; D013217:Starvation / Q000276:immunology / Q000503:physiopathology*
chemical_list: D051017:Apoptosis Regulatory Proteins; D001426:Bacterial Proteins; D008869:Microtubule-Associated Proteins; D020123:Sirolimus