Incorporation of tubulin from an evolutionarily diverse source, Physarum polycephalum, into the microtubules of a mammalian cell.
Physarum myxamoebal tubulin was injected into PtK2 cells to determine whether tubulin from this eukaryotic microbe could act as a reporter for microtubule growth and dynamics in a mammalian cell. The distribution of Physarum tubulin was determined by the use of a monoclonal antibody specific for Physarum tubulin and unable to detect mammalian tubulin. Physarum tubulin was incorporated into the microtubules of both interphase arrays and the mitotic spindle. Measurements of microtubule turnover kinetics were found to be similar to those of other studies in which chemically modified brain tubulin has been used. Results using this heterologous system demonstrate that tubulin from an evolutionarily diverse organism can be used as a marker for microtubule growth in mammalian cells. Furthermore, the Physarum tubulin was able to endow the injected cells with novel properties. Resistance to colchicine-induced microtubule disassembly, a characteristic of Physarum tubulin, was conferred on the injected PtK2 cells. Use of this heterologous reporter tubulin system has also revealed features of variation in microtubule dynamics both within individual cells and between cells.
Prescott
A R
AR
School of Biological Sciences, University of East Anglia, Norwich, UK.
Foster
K E
KE
Warn
R M
RM
Gull
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D005075:Biological Evolution; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D007700:Kinetics; D008845:Microinjections; D008854:Microscopy, Electron; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D008938:Mitosis; D015739:Nocodazole / Q000494:pharmacology; D010804:Physarum; D014404:Tubulin / Q000032:analysis / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D014404:Tubulin; D015739:Nocodazole; D003078:Colchicine