Leishmania adaptor protein-1 subunits are required for normal lysosome traffic, flagellum biogenesis, lipid homeostasis, and adaptation to temperatures encountered in the mammalian host.
The adaptor protein-1 (AP-1) complex is involved in membrane transport between the Golgi apparatus and endosomes. In the protozoan parasite Leishmania mexicana mexicana, the AP-1 mu1 and sigma1 subunits are not required for growth at 27 degrees C but are essential for infectivity in the mammalian host. In this study, we have investigated the function of these AP-1 subunits in order to understand the molecular basis for this loss of virulence. The mu1 and sigma1 subunits were localized to late Golgi and endosome membranes of the major parasite stages. Parasite mutants lacking either AP-1 subunit lacked obvious defects in Golgi structure, endocytosis, or exocytic transport. However, these mutants displayed reduced rates of endosome-to-lysosome transport and accumulated fragmented, sterol-rich lysosomes. Defects in flagellum biogenesis were also evident in nondividing promastigote stages, and this phenotype was exacerbated by inhibitors of sterol and sphingolipid biosynthesis. Furthermore, both AP-1 mutants were hypersensitive to elevated temperature and perturbations in membrane lipid composition. The pleiotropic requirements for AP-1 in membrane trafficking and temperature stress responses explain the loss of virulence of these mutants in the mammalian host.
Department of Biochemistry and Molecular Biology, University of Melbourne, Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Rd, Parkville, Victoria 3010, Australia.
James E
JE
Vince
Dedreia L
DL
Tull
Timothy
T
Spurck
Merran C
MC
Derby
Geoffrey I
GI
McFadden
Paul A
PA
Gleeson
Suzanne
S
Gokool
Malcolm J
MJ
McConville
D000222:Adaptation, Physiological / Q000502:physiology; D033961:Adaptor Protein Complex 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001831:Body Temperature / Q000502:physiology; D011992:Endosomes / Q000378:metabolism / Q000648:ultrastructure; D005407:Flagella / Q000378:metabolism* / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000378:metabolism / Q000648:ultrastructure; D018869:Heat-Shock Response / Q000502:physiology; D006706:Homeostasis / Q000502:physiology; D006790:Host-Parasite Interactions / Q000502:physiology*; D007894:Leishmania mexicana / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D050356:Lipid Metabolism / Q000502:physiology*; D008247:Lysosomes / Q000378:metabolism* / Q000648:ultrastructure; D008322:Mammals / Q000469:parasitology / Q000502:physiology; D008563:Membrane Lipids / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D013261:Sterols / Q000378:metabolism
D033961:Adaptor Protein Complex 1; D008563:Membrane Lipids; D021122:Protein Subunits; D013261:Sterols