Characterization of the interaction between Toxoplasma gondii rhoptry neck protein 4 and host cellular β-tubulin.
Toxoplasma rhoptry neck protein 4 (TgRON4) is a component of the moving junction macromolecular complex that plays a central role during invasion. TgRON4 is exposed on the cytosolic side of the host cell during invasion, but its molecular interactions remain unclear. Here, we identified host cellular β-tubulin as a binding partner of TgRON4, but not Plasmodium RON4. Coimmunoprecipitation studies in mammalian cells demonstrated that the C-terminal 15-kDa region of β-tubulin was sufficient for binding to TgRON4, and that a 17-kDa region in the proximal C-terminus of TgRON4 was required for binding to the C-terminal region of β-tubulin. Analysis of T. gondii-infected lysates from CHO cells expressing the TgRON4-binding region showed that the C-terminal region of β-tubulin interacted with TgRON4 at early invasion step. Our results provide evidence for a parasite-specific interaction between TgRON4 and the host cell cytoskeleton in parasite-infected cells.
1] National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan [2] Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Hitoshi
H
Takemae
Tatsuki
T
Sugi
Kyousuke
K
Kobayashi
Haiyan
H
Gong
Akiko
A
Ishiwa
Frances C
FC
Recuenco
Fumi
F
Murakoshi
Tatsuya
T
Iwanaga
Atsuko
A
Inomata
Taisuke
T
Horimoto
Hiroomi
H
Akashi
Kentaro
K
Kato
D000818:Animals; D000953:Antigens, Protozoan; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D006790:Host-Parasite Interactions / Q000502:physiology*; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism*; D014122:Toxoplasma / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*
D000953:Antigens, Protozoan; D002352:Carrier Proteins; D008565:Membrane Proteins; D015800:Protozoan Proteins; D014404:Tubulin; D010447:Peptide Hydrolases; C560858:ROM4 protein, Toxoplasma gondii