The IC138 and IC140 intermediate chains of the I1 axonemal dynein complex bind directly to tubulin.
Dyneins are minus end directed microtubule motors that play a critical role in ciliary and flagellar movement. Ciliary dyneins, also known as axonemal dyneins, are characterized based on their location on the axoneme, either as outer dynein arms or inner dynein arms. The I1 dynein is the best-characterized subspecies of the inner dynein arms; however the interactions between many of the components of the I1 complex and the axoneme are not well defined. In an effort to elucidate the interactions in which the I1 components are involved, we performed zero-length crosslinking on axonemes and studied the crosslinked products formed by the I1 intermediate chains, IC138 and IC140. Our data indicate that IC138 and IC140 bind directly to microtubules. Mass-spectrometry analysis of the crosslinked product identified both α- and β-tubulin as the IC138 and IC140 binding partners. This was further confirmed by crosslinking experiments carried out on purified I1 fractions bound to Taxol-stabilized microtubules. Furthermore, the interaction between IC140 and tubulin is lost when IC138 is absent. Our studies support previous findings that intermediate chains play critical roles in the assembly, axonemal targeting and regulation of the I1 dynein complex.
Department of Biology, Morehouse College, Atlanta, GA 30314, USA. Electronic address: thendrickson@morehouse.edu.
Triscia W
TW
Hendrickson
Department of Biology, Morehouse College, Atlanta, GA 30314, USA.
Jonathan L
JL
Goss
Department of Biology, Morehouse College, Atlanta, GA 30314, USA; Department of Chemistry, Washington University, St. Louis, MO 63130, USA.
Charles A
CA
Seaton
Department of Chemistry, Washington University, St. Louis, MO 63130, USA.
Henry W
HW
Rohrs
D000255:Adenosine Triphosphate / Q000494:pharmacology; D054468:Axoneme / Q000187:drug effects / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000187:drug effects / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004398:Dyneins / Q000378:metabolism*; D017239:Paclitaxel / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry; D010940:Plant Proteins / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D014404:Tubulin / Q000378:metabolism*
D003432:Cross-Linking Reagents; D010455:Peptides; D010940:Plant Proteins; D014404:Tubulin; D000255:Adenosine Triphosphate; D004398:Dyneins; D017239:Paclitaxel
Axoneme; Flagellum; I1 dynein; Tubulin; cilia