Leishmania major: differential regulation of the surface metalloprotease in amastigote and promastigote stages.
During its life cycle, the protozoan parasite Leishmania major alternates from an intracellular amastigote form in the mammalian host to a flagellated promastigote form in the insect vector. The expression of the surface metalloprotease (PSP) during differentiation in vitro was investigated by Western and Northern blots, by immunoprecipitation of cells metabolically labeled with [35S]methionine or labeled at the surface with radioactive iodine, and by quantification of the proteolytic activity in substrate-containing polyacrylamide gels. We report that the surface metalloprotease is down-regulated at both the mRNA and the protein level in amastigotes, where it represents less than 1% of the equivalent proteolytic activity detected in promastigotes. A significant amount of mRNA is detected 4 hr after the onset of differentiation. The expression of the protease begins at that time and reaches steady state 8 hr later. The synthesis of PSP precedes the complete morphological differentiation to the promastigote stage and the appearance of the lipophosphoglycan, another major promastigote surface component. In contrast to PSP, a family of mercaptoethanol-activated proteases present in the amastigote exists only at a reduced level in the promastigote. The confinement of the surface metalloprotease to the insect stage of the parasite suggests that it has no physiological function in the parasitism maintenance of mammalian host macrophages.
Schneider
P
P
Institut de Biochimie, WHO Immunology Research and Training Center, Université de Lausanne, Epalinges, Switzerland.
Rosat
J P
JP
Bouvier
J
J
Louis
J
J
Bordier
C
C
D000818:Animals; D015536:Down-Regulation; D010450:Endopeptidases / Q000096:biosynthesis; D015971:Gene Expression Regulation, Enzymologic; D006028:Glycosphingolipids / Q000096:biosynthesis; D007895:Leishmania tropica / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism
D006028:Glycosphingolipids; D012333:RNA, Messenger; D016053:RNA, Protozoan; C008290:lipophosphonoglycan; D010450:Endopeptidases; D008666:Metalloendopeptidases; C055146:glycoprotein gp63, Leishmania