Unique functional characteristics of the polymerization and MAP binding regulatory domains of plant tubulin.
An understanding of the regulation of microtubule polymerization and dynamics in plant cells requires biochemical information on the structures, functions, and molecular interactions of plant tubulin and microtubule-associated proteins (MAPs) that regulate microtubule function. We have probed the regulatory domain and polymerization domain of purified maize tubulin using MAP2, an extensively characterized mammalian neuronal MAP. MAP2 bound to the surface of preformed, taxol-stabilized maize microtubules, with binding saturation occurring with one MAP2 molecule per five to six tubulin dimers, as it does with mammalian microtubules. MAP2 binding and dissociation analyses revealed two affinity classes of binding sites on maize microtubules: a high-affinity site 12 dimers apart that may be homologous to the mammalian MAP2 binding site and an additional low-affinity site also 12 dimers apart that may be homologous to the mammalian tau binding site. MAP2 corrected in vitro folding errors in taxol-stabilized maize microtubules and reduced the critical concentration of maize tubulin polymerization eightfold, from 8.3 to 1.0 microM. However, MAP2 dissociated much more readily from maize microtubules than from mammalian microtubules and induced the assembly of maize tubulin into aberrant helical ribbon polymers that remained stable for prolonged periods. Our results indicated that MAP2 binds to maize tubulin via a partially specific, low-fidelity interaction that reflects unique structural and functional properties of the polymerization and regulatory domains of plant tubulin and possibly of the tubulin binding domains of undocumented MAPs that regulate microtubule function in plant cells.
Hugdahl
J D
JD
Department of Botany, University of Texas at Austin 78713.
Bokros
C L
CL
Hanesworth
V R
VR
Aalund
G R
GR
Morejohn
L C
LC
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001704:Biopolymers / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002417:Cattle; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D017239:Paclitaxel / Q000494:pharmacology; D011485:Protein Binding / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014404:Tubulin / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D003313:Zea mays / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
D001704:Biopolymers; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D017239:Paclitaxel