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A more recent version of this article appeared on July 1, 2002
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Submitted on January 3, 2002
Revised on March 21, 2002
Accepted on March 25, 2002
-tubulin complex, Alp16, is required for cytoplasmic microtubule integrity and becomes indispensable when -tubulin function is compromised
1 Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, UK
2 Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, UK (present address: Centro de Biología Molecular "Severo Ochoa", Universidad Autónoma de Madrid, 28049 Cantoblanco, Madrid, Spain)
* Corresponding author. E-mail address: toda{at}cancer.org.uk.
-Tubulin functions as a multiprotein complex, called the -tubulin complex (-TuC) and comprises the microtubule organising centre (MTOC). Fission yeast Alp4 and Alp6 are homologues of two conserved -tubulin complex proteins hGCP2 and hGCP3 respectively. We isolated a novel gene alp16+ as a multicopy suppressor of temperature-sensitive (ts) alp6-719 mutants. alp16+ encodes a 759 amino acid protein with two conserved regions found in all other members of -TuC components. In addition, Alp16 contains an additional motif, which shows homology to hGCP6/Xgrip210. Gene disruption shows that alp16+ is not essential for cell viability. However, alp16-deletion displays abnormally long cytoplasmic microtubules, which curve around the cell tip. Furthermore, alp16-deleted mutants are hypersensitive to microtubule-depolymerising drugs and synthetically lethal with either of ts alp4-225, alp4-1891 or alp6-719 mutants. Overproduction of Alp16 is lethal with defective phenotypes very similar to loss of Alp4 or Alp6. Alp16 localises to the spindle pole body (SPB) throughout the cell cycle and to the equatorial MTOC at post-anaphase. Alp16 co-immunoprecipitates with -tubulin and co-sediments with the -TuC in a large complex (>20 S). Alp16 is, however, not required for the formation of this large complex. We discuss evolutional conservation and divergence of structure and function of the -TuC between yeast and higher eukaryotes.
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