Subcellular localization of the human proto-oncogene protein DEK

被引:88
作者
Kappes, F [1 ]
Burger, K [1 ]
Baack, M [1 ]
Fackelmayer, FO [1 ]
Gruss, C [1 ]
机构
[1] Univ Konstanz, Dept Biol, D-78457 Constance, Germany
关键词
D O I
10.1074/jbc.M100162200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Recent data revealed that DEK associates with splicing complexes through interactions mediated by serine/arginine-repeat proteins. However, the DEK protein has also been shown to change the topology of DNA in chromatin in vitro, This could indicate that the DEK protein resides on cellular chromatin, To investigate the in vivo localization of DEK, we performed cell fractionation studies, immunolabeling, and micrococcal nuclease digestion analysis. Most of the DEK protein was found to be released by DNase treatment of nuclei, and only a small amount by treatment with RNase, Furthermore, micrococcal nuclease digestion of nuclei followed by glycerol gradient sedimentation revealed that DEK cosedimentates with oligonucleosomes, clearly demonstrating that DEK is associated with chromatin in vivo. Additional chromatin fractionation studies, based on the different accessibilities to micrococcal nuclease, showed that DEK is associated both with extended, genetically active and more densely organized, inactive chromatin, We found no significant change in the amount and localization of DEK in cells that synchronously traversed the cell cycle. In summary these data demonstrate that the major portion of DEK is associated with chromatin in vivo and suggest that it might play a role in chromatin architecture.
引用
收藏
页码:26317 / 26323
页数:7
相关论文
共 40 条
  • [1] ADAM SA, 1992, METHOD ENZYMOL, V219, P97
  • [2] Alexiadis V, 2000, GENE DEV, V14, P1308
  • [3] SAP - a putative DNA-binding motif involved in chromosomal organization
    Aravind, L
    Koonin, EV
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 2000, 25 (03) : 112 - 114
  • [4] BLENCOWE BJ, 1995, RNA, V1, P852
  • [5] STRUCTURE OF CHROMATIN
    CLARK, RJ
    FELSENFELD, G
    [J]. NATURE-NEW BIOLOGY, 1971, 229 (04): : 101 - +
  • [6] WT1 interacts with the splicing factor U2AF65 in an isoform-dependent manner and can be incorporated into spliceosomes
    Davies, RC
    Calvio, C
    Bratt, E
    Larsson, SH
    Lamond, AI
    Hastie, ND
    [J]. GENES & DEVELOPMENT, 1998, 12 (20) : 3217 - 3225
  • [7] Dong XW, 1998, ARTHRITIS RHEUM-US, V41, P1505, DOI 10.1002/1529-0131(199808)41:8<1505::AID-ART23>3.0.CO
  • [8] 2-N
  • [9] FORNEROD M, 1995, ONCOGENE, V10, P1739
  • [10] DEK, an autoantigen involved in a chromosomal translocation in acute myelogenous leukemia, binds to the HIV-2 enhancer
    Fu, GK
    Grosveld, G
    Markovitz, DM
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (05) : 1811 - 1815