The use of nocodazole in cell cycle analysis and parasite purification from Theileria parva-infected B cells.
Theileria parasites transform bovine leukocytes and induce uncontrolled lymphoproliferation only in the macroschizont stage of their life cycle. The isolation of highly purified stage-specific parasite RNA and proteins is an essential prerequisite when studying the Theileria-host relationship. We therefore improved a protocol based on the cytolytic bacterial toxin aerolysin by taking advantage of the microtubule inhibitor nocodazole. In this report we describe that nocodazole-mediated separation of the parasite from the host cell microtubule network was used with success to improve quantity and quality of purified parasites. We furthermore show that nocodazole is a useful tool to study cell cycle checkpoints due to its capacity to induce reversible cell cycle arrest in Theileria-infected B cells.
Baumgartner
M
M
Unité de biologie des interactions hôte-parasite, Institut Pasteur, Paris, France.
Tardieux
I
I
Ohayon
H
H
Gounon
P
P
Langsley
G
G
D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects / Q000469:parasitology*; D001427:Bacterial Toxins / Q000494:pharmacology; D002417:Cattle; D002453:Cell Cycle / Q000187:drug effects*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000469:parasitology; D006460:Hemolysin Proteins / Q000494:pharmacology; D015151:Immunoblotting; D008854:Microscopy, Electron; D008870:Microtubules / Q000187:drug effects / Q000469:parasitology; D015739:Nocodazole / Q000494:pharmacology*; D052899:Pore Forming Cytotoxic Proteins; D011505:Protein-Tyrosine Kinases / Q000032:analysis; D011518:Proto-Oncogene Proteins / Q000032:analysis; D051572:Proto-Oncogene Proteins c-hck; D016797:Theileria parva / Q000302:isolation & purification* / Q000648:ultrastructure; D013997:Time Factors
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D052899:Pore Forming Cytotoxic Proteins; D011518:Proto-Oncogene Proteins; C032676:aerolysin; D011505:Protein-Tyrosine Kinases; D051572:Proto-Oncogene Proteins c-hck; D015739:Nocodazole