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Plant Cell, Vol. 12, 1719-1736, September 2000, Copyright © 2000, American Society of Plant Physiologists

Identification and Characterization of a Novel Microtubule-Based Motor Associated with Membranous Organelles in Tobacco Pollen Tubes

Giampiero Caia, Silvia Romagnolia, Alessandra Moscatellib, Elisa Ovidic, Gabriella Gambellinic, Antonio Tiezzic, and Mauro Crestia
a Dipartimento di Scienze Ambientali, Università di Siena, via Mattioli 4, 53100 Siena, Italy
b Istituto di Botanica, Università di Messina, Salita Sperone 31, Contrada Papardo, Messina, Italy
c Dipartimento Scienze Ambientali, Università della Tuscia, via San Camillo de Lellis, Viterbo, Italy

Correspondence to: Giampiero Cai, cai{at}unisi.it (E-mail), 39-0577-232860 (fax)

Pollen tube growth depends on the differential distribution of organelles and vesicles along the tube. The role of microtubules in organelle movement is uncertain, mainly because information at the molecular level is limited. In an effort to understand the molecular basis of microtubule-based movement, we isolated from tobacco pollen tubes polypeptides that cosediment with microtubules in an ATP-dependent manner. Major polypeptides released from microtubules by ATP (ATP-MAPs) had molecular masses of 90, 80, and 41 kD. Several findings indicate that the 90-kD ATP-MAP is a kinesin-related motor: binding of the polypeptide to microtubules was enhanced by the nonhydrolyzable ATP analog AMP-PNP; the 90-kD polypeptide reacted specifically with a peptide antibody directed against a highly conserved region in the motor domain of the kinesin superfamily; purified 90-kD ATP-MAP induced microtubules to glide in motility assays in vitro; and the 90-kD ATP-MAP cofractionated with microtubule-activated ATPase activity. Immunolocalization studies indicated that the 90-kD ATP-MAP binds to organelles associated with microtubules in the cortical region of the pollen tube. These findings suggest that the 90-kD ATP-MAP is a kinesin-related microtubule motor that moves organelles in the cortex of growing pollen tubes.




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