ESSENTIAL ROLE OF PROTEIN-PHOSPHORYLATION IN NUCLEAR TRANSPORT

被引:26
作者
MISHRA, K [1 ]
PARNAIK, VK [1 ]
机构
[1] CTR CELLULAR & MOLEC BIOL, HYDERABAD 500007, ANDHRA PRADESH, INDIA
关键词
D O I
10.1006/excr.1995.1016
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
We have investigated a possible role for protein phosphorylation in nuclear transport in semi-intact cells, prepared by digitonin permeabilization of rat F-111 fibroblasts. Treatment of semi-intact cells with alkaline phosphatase abolished the import of nuclear transport substrates, namely, signal peptide-albumin conjugates, as well as their signal-dependent binding at the nuclear pores, but did not affect the morphology of the cells, in particular their cytoskeletal network. Authentic transport and functional binding of the karyophilic protein at the nuclear envelope could be restored by incubation of phosphatase-treated cells with cytosol enriched in protein kinase C or with purified protein kinase A (catalytic subunit). Restoration of transport was blocked by specific inhibitors of these kinases. Since the protein phosphorylation required for nuclear transport appeared to be a reasonably stable modification, characterization of the phosphorylated proteins was attempted in kinase reactions with radiolabeled ATP. Two proteins of 60-62 kDa were the predominant substrates phosphorylated by both protein kinase C and protein kinase A under conditions wherein nuclear transport was restored. Our results suggest a requirement for phosphorylation of one or more proteins for binding of a karyophilic protein at the nuclear envelope. (C) 1995 Academic Press,Inc.
引用
收藏
页码:124 / 134
页数:11
相关论文
共 44 条
  • [11] STRUCTURE AND FUNCTION OF THE NUCLEAR-PORE COMPLEX
    FORBES, DJ
    [J]. ANNUAL REVIEW OF CELL BIOLOGY, 1992, 8 : 495 - 527
  • [12] NUCLEAR-PROTEIN LOCALIZATION
    GARCIABUSTOS, J
    HEITMAN, J
    HALL, MN
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1071 (01) : 83 - 101
  • [13] FUNCTIONAL-ORGANIZATION OF THE NUCLEAR-ENVELOPE
    GERACE, L
    BURKE, B
    [J]. ANNUAL REVIEW OF CELL BIOLOGY, 1988, 4 : 335 - 374
  • [14] SYNTHETIC PEPTIDES AS NUCLEAR-LOCALIZATION SIGNALS
    GOLDFARB, DS
    GARIEPY, J
    SCHOOLNIK, G
    KORNBERG, RD
    [J]. NATURE, 1986, 322 (6080) : 641 - 644
  • [15] NUCLEAR-PROTEIN IMPORT IS INHIBITED BY AN ANTIBODY TO A LUMENAL EPITOPE OF A NUCLEAR-PORE COMPLEX GLYCOPROTEIN
    GREBER, UF
    GERACE, L
    [J]. JOURNAL OF CELL BIOLOGY, 1992, 116 (01) : 15 - 30
  • [16] AN INTEGRAL MEMBRANE-PROTEIN OF THE PORE MEMBRANE DOMAIN OF THE NUCLEAR-ENVELOPE CONTAINS A NUCLEOPORIN-LIKE REGION
    HALLBERG, E
    WOZNIAK, RW
    BLOBEL, G
    [J]. JOURNAL OF CELL BIOLOGY, 1993, 122 (03) : 513 - 521
  • [17] HIDAKA H, 1991, METHOD ENZYMOL, V201, P328
  • [18] ANTIBODIES AGAINST 70-KD HEAT-SHOCK COGNATE PROTEIN INHIBIT MEDIATED NUCLEAR IMPORT OF KARYOPHILIC PROTEINS
    IMAMOTO, N
    MATSUOKA, Y
    KURIHARA, T
    KOHNO, K
    MIYAGI, M
    SAKIYAMA, F
    OKADA, Y
    TSUNASAWA, S
    YONEDA, Y
    [J]. JOURNAL OF CELL BIOLOGY, 1992, 119 (05) : 1047 - 1061
  • [19] A CONVENIENT METHOD FOR THE SYNTHESIS OF PEPTIDES ACYLATED WITH PALMITIC ACID SPECIFICALLY AT CYSTEINE THIOL
    JOSEPH, M
    NAGARAJ, R
    [J]. BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 1993, 3 (06) : 1025 - 1028
  • [20] KAMPS MP, 1991, METHOD ENZYMOL, V201, P21