Yeast proteins associated with microtubules in vitro and in vivo.
Conditions were established for the self-assembly of milligram amounts of purified Saccharomyces cerevisiae tubulin. Microtubules assembled with pure yeast tubulin were not stabilized by taxol; hybrid microtubules containing substoichiometric amounts of bovine tubulin were stabilized. Yeast microtubule-associated proteins (MAPs) were identified on affinity matrices made from hybrid and all-bovine microtubules. About 25 yeast MAPs were isolated. The amino-terminal sequences of several of these were determined: three were known metabolic enzymes, two were GTP-binding proteins (including the product of the SAR1 gene), and three were novel proteins not found in sequence databases. Affinity-purified antisera were generated against synthetic peptides corresponding to two of the apparently novel proteins (38 and 50 kDa). Immunofluorescence microscopy showed that both these proteins colocalize with intra- and extranuclear microtubules in vivo.
Barnes
G
G
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Louie
K A
KA
Botstein
D
D
D000470:Alkaloids / Q000494:pharmacology; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008870:Microtubules / Q000737:chemistry*; D008970:Molecular Weight; D017239:Paclitaxel; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D014404:Tubulin / Q000737:chemistry* / Q000302:isolation & purification
D000470:Alkaloids; D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D019204:GTP-Binding Proteins; D017239:Paclitaxel