Dissecting the involvement of formins in Bud6p-mediated cortical capture of microtubules in S. cerevisiae.

In S. cerevisiae, spindle orientation is linked to the inheritance of the ;old' spindle pole by the bud. A player in this asymmetric commitment, Bud6p, promotes cortical capture of astral microtubules. Additionally, Bud6p stimulates actin cable formation though the formin Bni1p. A relationship with the second formin, Bnr1p, is unclear. Another player is Kar9p, a protein that guides microtubules along actin cables organised by formins. Here, we ask whether formins mediate Bud6p-dependent microtubule capture beyond any links to Kar9p and actin. We found that both formins control Bud6p localisation. bni1 mutations advanced recruitment of Bud6p at the bud neck, ahead of spindle assembly, whereas bnr1Delta reduced Bud6p association with the bud neck. Accordingly, bni1 or bnr1 mutations redirected microtubule capture to or away from the bud neck, respectively. Furthermore, a Bni1p truncation that can form actin cables independently of Bud6p could not bypass a bud6Delta for microtubule capture. Conversely, Bud6(1-565)p, a truncation insufficient for correct actin organisation via formins, supported microtubule capture. Finally, Bud6p or Bud6(1-565)p associated with microtubules in vitro. Thus, surprisingly, Bud6p may promote microtubule capture independently of its links to actin organisation, whereas formins would contribute to the program of Bud6p-dependent microtubule-cortex interactions by controlling Bud6p localisation.