Dynein is a transient kinetochore component whose binding is regulated by microtubule attachment, not tension

Cytoplasmic dynein is the only known kinetochore protein capable of driving chromosome movement toward spindle poles. In grasshopper spermatocytes, dy nein immunofluorescence staining is bright at prometaphase kinetochores and dimmer at metaphase kinetochores. We have determined that these difference s in staining intensity reflect differences in amounts of dynein associated with the kinetochore. Metaphase kinetochores regain bright dynein staining if they are detached from spindle microtubules by micromanipulation and ke pt detached for 10 min. We show that this increase in dynein staining is no t caused by the retraction or unmasking of dynein upon detachment. Thus, dy nein genuinely is a transient component of spermatocyte kinetochores. We further show that microtubule attachment, not tension, regulates dynein localization at kinetochores. Dynein binding is extremely sensitive to the presence of microtubules: fewer than half the normal number of kinetochore microtubules leads to the loss of most kinetochoric dynein, As a result, th e bulk of the dynein leaves the kinetochore very early in mitosis, soon aft er the kinetochores begin to attach to microtubules. The possible functions of this dynein fraction are therefore limited to the initial attachment an d movement of chromosomes and/or to a role in the mitotic checkpoint.