Sep, 1999
Functional dissection and hierarchy of tubulin-folding cofactor homologues in fission yeast
MOLECULAR BIOLOGY OF THE CELL
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- Volume
- 10
- Number
- 9
- First page
- 2987
- Last page
- 3001
- Language
- English
- Publishing type
- Publisher
- AMER SOC CELL BIOLOGY
We describe the isolation of fission yeast homologues of tubulin-folding cofactors B (Alp11) and E (Alp21), which are essential for cell viability and the maintenance of microtubules. Alp11B contains the glycine-rich motif (the CLIP-170 domain) involved in microtubular functions, whereas, unlike mammalian cofactor E, Alp21(E) does not. both mammalian and yeast cofactor E, however, do contain leucine-rich repeats. Immunoprecipitation analysis shows that Alp11(B) interacts with both a-tubulin and Alp21(E), but not with the cofactor D homologue Alp1, whereas Alp21(E) also interacts with Alpl(D). The cellular amount of a-tubulin is decreased in both alp1 and alp(11) mutants. Overproduction of Alp11(B) results in cell lethality and the disappearance of microtubules, which is rescued by co-overproduction of alpha-tubulin. Both hull-length Alp11(B) and the C-terminal third containing the CLIP-170 domain localize in the cytoplasm, and this domain is required for efficient binding to alpha-tubulin. Deletion of alp11 is suppressed by multicopy plasmids containing either alp21(+) or alp(1+), whereas alp(21) deletion is rescued by overexpression of either alp11(+) or alp21(+). The alp1(+) but not alp11(+). Finally, the alp1 mutant is not complemented by either alp11(+) or alp21(+). The results suggest that cofactors operate in a linear pathway (Alp11(B)-Alp21(E)-Alp(D)), each with distinct roles.
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- ID information
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- ISSN : 1059-1524
- eISSN : 1939-4586
- Web of Science ID : WOS:000082598500015