GAMMA-TUBULIN IS REQUIRED FOR THE STRUCTURE AND FUNCTION OF THE MICROTUBULE-ORGANIZING CENTER IN DROSOPHILA NEUROBLASTS

We report that in Drosophila, gamma-tubulin is required for the structure as well as the function of microtubule organizing centres (MTOCs). This conclusion is based on the identification and phenotypic characterization of a mutant allele of the gamma-tubulin gene located at region 23C of the polytene chromosome map. This mutation, which we have called gamma-tub23C(PI), is caused by the insertion of a P-element within the 5' untranslated leader of the gamma-tubulin transcript. Northern and Western analyses show that gamma-tub23C(PI) is either a null or a very severe hypomorph as no gamma-tubulin mRNA or protein can be detected in mutant individuals. Visualization of DNA, MTOCs and microtubules by confocal laser scanning microscopy of cells from individuals homozygous for gamma-tub23C(PI) reveals a series of phenotypic abnormalities. Some of these are similar to those observed after disruption of gamma-tubulin function in other organisms, including mitotic arrest and a dramatic decrease in the number of microtubules, but, in addition, we have observed that mutation in this gene also results in highly abnormal MTOCs which show a variety of shapes and sizes which we never observed in wild type cells. These results show that gamma-tubulin is required for both structural and functional roles in the MTOCs.