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Volumn 287, Issue 5461, 2000, Pages 2260-2262

Positioning of the mitotic spindle by a cortical-microtubule capture mechanism

Author keywords

[No Author keywords available]

Indexed keywords

GREEN FLUORESCENT PROTEIN; HYBRID PROTEIN; PROTEIN;

EID: 0034708606     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.287.5461.2260     Document Type: Article
Times cited : (244)

References (40)
  • 23
    • 0343687478 scopus 로고    scopus 로고
    • Supplemental material is available at www. sciencemag.org/feature/data/1047337.shl
  • 26
    • 0030927327 scopus 로고    scopus 로고
    • H. Imamura et al., EMBO J. 16, 2745 (1997).
    • (1997) EMBO J. , vol.16 , pp. 2745
    • Imamura, H.1
  • 28
    • 0343251808 scopus 로고    scopus 로고
    • note
    • The delivery of Bim1p to cortical Kar9p binding sites could signal the successful completion of spindle positioning and promote cytokinesis on schedule. However, we found that Kar9p was not required for the cytokinesis checkpoint (L. Lee and D. Pellman, unpublished data).
  • 30
  • 33
    • 0343687475 scopus 로고    scopus 로고
    • note
    • An astral microtubule was considered correctly oriented if it extended into the bud or mating projection. Analysis was limited to microtubules of length equal to or greater than the shortest distance from the centrosome to the GFP-Kar9p signal. The percentage of preanaphase cells that met the above criteria was 89% in the BIM1 strain (n = 100) and 8% in the bim1Δ strain (n = 228). In α factor-arrested cells, the percentage was 89% in the BIM1 strain (n = 141) and 27% in the bim1Δ strain (n = 321). It is likely that misoriented microtubules in bim1Δ cells are misoriented because they fail to interact with Kar9p. If this is the case, then Bim1p is required for microtubule ends to interact with Kar9p in most cells.
  • 34
    • 0342817243 scopus 로고    scopus 로고
    • note
    • Cell extracts for Western analysis and immunoprecipitations were prepared by glass bead lysis in a buffer containing 150 mM NaCl, 50 mM Tris (pH 8.0), 1% Nonidet P-40, and protease inhibitors. Antibodies for immunoblotting: HA-Bim1p was detected with 12CA5, GFP-Kar9p was detected with a polydonal antibody to GFP, and tubulin was detected with a monoclonal antibody to α tubulin.
  • 35
    • 0342817242 scopus 로고    scopus 로고
    • note
    • Sucrose gradients (5 to 20%) were made in 50 mM Tris (pH 8.0) and 150 mM NaCl. We loaded 400 μl of low-speed supernatant on top of each 12.5-ml gradient and spun it overnight in a Beckman SW40 Ti at 218,000g. The linearity of the gradients was confirmed by refractometry, and Svedberg values were calculated on the basis of globular standards. Gel filtration was performed using a Bio-Rad SE-1000 analytical Gel Filtration column.
  • 40
    • 0343687474 scopus 로고    scopus 로고
    • note
    • We thank J. DeCaprio, E. Elion, J. Kahana, R. Miller, M. Rose, and P. Silver for reagents, E. Elion, R. Li, T. Look, and M. McLaughlin for comments on the manuscript, J. Chant and M. Rose for communicating results before publication, and B. Bierer and members of the Pellman laboratory for discussions. Supported by grants from NIH [GM55772 to D.P. and KO8 DK02578 to J.S.T.] and a Kimmel Scholar Award to D.P.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.