Distinct Dgrip84 isoforms correlate with distinct gamma-tubulins in Drosophila

gamma-Tubulin is an indispensable component of the animal centrosome and is required for proper microtubule organization. Within the cell, gamma-tubulin exists in a multiprotein complex containing between two (some yeasts) and six or more (metazoa) additional highly conserved proteins named gamma ring proteins (Grips) or gamma complex proteins (GCPs). gamma-Tubulin containing complexes isolated from Xenopus eggs or Drosophila embryos appear ring-shaped and have therefore been named the gamma-tubulin ring complex (gamma TuRC). Curiously, many organisms (including humans) have two distinct gamma-tubulin genes. In Drosophila, where the two gamma-tubulin isotypes have been studied most extensively, the gamma-tubulin genes are developmentally regulated: the maternal gamma-tubulin isotype ( named gamma Tub37CD according to its location on the genetic map) is expressed in the ovary and is deposited in the egg, where it is thought to orchestrate the meiotic and early embryonic cleavages. The second gamma-tubulin isotype (gamma Tub23C) is ubiquitously expressed and persists in most of the cells of the adult fly. In those rare cases where both gamma-tubulins coexist in the same cell, they show distinct subcellular distributions and cell-cycle-dependent changes: gamma Tub37CD mainly localizes to the centrosome, where its levels vary only slightly with the cell cycle. In contrast, the level of gamma Tub23C at the centrosome increases at the beginning of mitosis, and gamma Tub23C also associates with spindle pole microtubules. Here, we show that gamma Tub23C forms discrete complexes that closely resemble the complexes formed by gamma Tub37CD. Surprisingly, however, gamma Tub23C associates with a distinct, longer splice variant of Dgrip84. This may reflect a role for Dgrip84 in regulating the activity and/or the location of the gamma-tubulin complexes formed with gamma Tub37CD and gamma Tub23C.