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Volumn 310, Issue 5753, 2005, Pages 1499-1504

Cell biology: Chromosome alignment and segregation regulated by ubiquitination of survivin

Author keywords

[No Author keywords available]

Indexed keywords

BIODEGRADATION; CHROMOSOMES; ENZYMES;

EID: 28544453333     PISSN: 00368075     EISSN: 10959203     Source Type: Journal    
DOI: 10.1126/science.1120160     Document Type: Article
Times cited : (215)

References (37)
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    • note
    • INS was made by changing wobble codons (GCA GTT AGC GGG TGG AAG, changed nucleotides are in bold) corresponding to hFAM-s1 siRNA sequence using GeneEditor (Promega). Antibody to Xenopus Survivin was raised against glutathione S-transferase (GST) Xenopus Survivin and affinity purified using cleaved Survivin. Antibodies against hFAM were raised against each of four peptides (TPPDEQGQG-DAPPQLED, CAPDEHESPPPEDAP, QRAQENYEG-SEEVSPPQTKDQ, and GDEKQDNESNVDPRDDV) (36) and one GST fusion of the C-terminal fragment (amino acids 2347 to 2547) of hFAM. All were affinity purified against the respective antigens. Mouse Ufd1 was cloned into the pEF6/V5-His vector. Ufd1 antibodies were raised against GST-Ufd1 fusion protein and purified against His-tagged Ufd1. Antibodies against tubulin (Sigma), ACA (Antibodies Incorporated), human Survivin (R&D systems), human Aurora B (BD Biosciences), HA (Roche), Myc (Santa Cruz), and control immunoglobulin G (IgG) (Jackson Laboratory) were purchased. Antibodies against phosphorylated MCAK were described previously (27). Immunoprecipitation and immunofluorescence were carried out as described previously (15).
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    • note
    • CAT, or pV5 vector for at least 48 hours before siRNA transfection. Cells were analyzed 72 hours after siRNA transfection.
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    • note
    • HeLa cells were treated with hFAM-s1 or control siRNA for ∼24 hours and then transfected with Survivin-GFP. FLIP or FRAP was carried out 48 to 72 hours after transfection. Detailed descriptions of the kinetic modeling of FLIP and FRAP can be found in (26).
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    • note
    • Materials and methods are available as supporting materials on Science Online.
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    • note
    • CAT or the respective vector controls followed by the same treatment and analyses described above.
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    • 28544443555 scopus 로고    scopus 로고
    • note
    • Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
  • 37
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    • note
    • We thank J. Swedlow, S. Wood, H. Meyer, and T. Dawson for antibodies and constructs B. Lane for protein sequencing; C. Pickart for helpful advice on ubiquitination assays; O. Martin and R. Chen for excellent technical support; and M. Guo, D. Koshland, J. Yanowitz, and the members of Zheng lab for helpful comments. This work was supported by Howard Hughes Medical Institution and by National Institute of General Medical Sciences grant no. GM56312.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.