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J. Biol. Chem., Vol. 275, Issue 21, 16281-16288, May 26, 2000
From the We investigated the intracellular route of
Salmonella in macrophages to determine a plausible
mechanism for their survival in phagocytes. Western blot analysis of
isolated phagosomes using specific antibodies revealed that by 5 min
after internalization dead Salmonella-containing phagosomes
acquire transferrin receptors (a marker for early endosomes), whereas
by 30 min the dead bacteria are found in vesicles carrying the late
endosomal markers cation-dependent mannose 6-phosphate
receptors, Rab7 and Rab9. In contrast, live Salmonella-containing phagosomes (LSP) retain a significant
amount of Rab5 and transferrin receptor until 30 min, selectively
deplete Rab7 and Rab9, and never acquire mannose 6-phosphate receptors even 90 min after internalization. Retention of Rab5 and Rab18 and
selective depletion of Rab7 and Rab9 presumably enable the LSP to avoid
transport to lysosomes through late endosomes. The presence of immature
cathepsin D (48 kDa) and selective depletion of the vacuolar ATPase in
LSP presumably contributes to the less acidic pH of LSP. In contrast,
proteolytically processed cathepsin D (Mr
17,000) was detected by 30 min on the dead
Salmonella-containing phagosomes. Morphological analysis
also revealed that after uptake by macrophages, the dead
Salmonella are transported to lysosomes, whereas the live
bacteria persist in compartments that avoid fusion with lysosomes,
indicating that live Salmonella bypass the normal endocytic
route targeted to lysosomes and mature in a specialized compartment.
Live Salmonella Modulate Expression of Rab Proteins
to Persist in a Specialized Compartment and Escape Transport to
Lysosomes*
,,, and¶
National Institute of Immunology, Aruna Asaf
Ali Marg, New Delhi 110067, India and the § Institute of
Microbial Technology, Sector 39A, Chandigarh 160036, India
*
This work was supported by grants from the Department of
Biotechnology, Indian Council of Medical Research (to A. M.) and by
funds from the Jawaharlal Nehru Center for Advanced Scientific Research, Bangalore (to S. K. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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