External GTP binding and induction of cell division in starved Tetrahymena thermophila.
The extracellular nucleotide, guanosine 5'-triphosphate (GTP) is known to be a chemorepellent for ciliated protozoa such as Paramecium and Tetrahymena. Here, we studied the surface localization of GTP binding sites and also its effects on the cell division of Tetrahymena thermophila. When a ribose-modified and fluorescent analog of GTP, 2'-(or -3')-O-trinitrophenyl (TNP)-GTP was added to the cells starved in non-nutrient buffer, a remarkable fluorescence was observed at the compound cilia of the oral area, while it was weak at other cilia and the somatic membrane. Following transfer of the cells to the starvation medium, the intensity of TNP-GTP fluorescence at the oral area gradually increased and was saturated at 3-4 hours. Addition of GTP to the starved cell induced not only an avoiding reaction in swimming, but also induced a synchronous cell division that occurred 2 hours later. An attempt to search for other stimuli, which induced cell division, revealed that mechanical stimulation by a short period of centrifugation was almost as effective as the addition of GTP. The supernatant after centrifugation had the ability to induce cell division, and such activity was abolished after the supernatant was treated with the phosphatase, apyrase, suggesting the release of GTP by the mechanical stimulation. These results indicate that the released GTP binds mainly to the oral area and this then induces the cell division of starved T. thermophila.
Iwamoto
Masaaki
M
Division of Biophysical Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Japan.
Nakaoka
Yasuo
Y
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000502:physiology*; D005453:Fluorescence; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D016808:Tetrahymena thermophila / Q000378:metabolism*
D006160:Guanosine Triphosphate; C045093:2',3'-O-(2,4,6-trinitrocyclohexadienylidene)guanosine 5'-triphosphate; D002118:Calcium