Modulation of Leishmania major aquaglyceroporin activity by a mitogen-activated protein kinase.
Leishmania major aquaglyceroporin (LmjAQP1) adventitiously facilitates the uptake of antimonite [Sb(III)], an active form of Pentostam® or Glucantime®, which are the first line of defence against all forms of leishmaniasis. The present paper shows that LmjAQP1 activity is modulated by the mitogen-activated protein kinase, LmjMPK2. Leishmania parasites coexpressing LmjAQP1 and LmjMPK2 show increased Sb(III) uptake and increased Sb(III) sensitivity. When subjected to a hypo-osmotic stress, these cells show faster volume recovery than cells expressing LmjAQP1 alone. LmjAQP1 is phosphorylated in vivo at Thr-197 and this phosphorylation requires LmjMPK2 activity. Lys-42 of LmjMPK2 is critical for its kinase activity. Cells expressing altered T197A LmjAQP1 or K42A LmjMPK2 showed decreased Sb(III) influx and a slower volume recovery than cells expressing wild-type proteins. Phosphorylation of LmjAQP1 led to a decrease in its turnover rate affecting LmjAQP1 activity. Although LmjAQP1 is localized to the flagellum of promastigotes, upon phosphorylation, it is relocalized to the entire surface of the parasite. Leishmania mexicana promastigotes with an MPK2 deletion showed reduced Sb(III) uptake and slower volume recovery than wild-type cells. This is the first report where a parasite aquaglyceroporin activity is post-translationally modulated by a mitogen-activated protein kinase.
Department of Molecular Microbiology and Infectious Diseases, Florida International University, Herbert Wertheim College of Medicine, Miami, FL, USA.
Goutam
G
Mandal
Mansi
M
Sharma
Martin
M
Kruse
Claudia
C
Sander-Juelch
Laura A
LA
Munro
Yong
Y
Wang
Jenny Veide
JV
Vilg
Markus J
MJ
Tamás
Hiranmoy
H
Bhattacharjee
Martin
M
Wiese
Rita
R
Mukhopadhyay
D000965:Antimony / Q000378:metabolism; D000981:Antiprotozoal Agents / Q000378:metabolism; D051398:Aquaporin 1 / Q000378:metabolism*; D017353:Gene Deletion; D018320:Leishmania major / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism*; D007894:Leishmania mexicana / Q000201:enzymology / Q000235:genetics; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D021261:Parasitic Sensitivity Tests
D000981:Antiprotozoal Agents; D051398:Aquaporin 1; C061374:antimonite; D000965:Antimony; D020928:Mitogen-Activated Protein Kinases