Phosphorylation of alpha B-crystallin in response to various types of stress

被引:178
作者
Ito, H
Okamoto, K
Nakayama, H
Isobe, T
Kato, K
机构
[1] AICHI HUMAN SERV CTR,INST DEV RES,DEPT BIOCHEM,KASUGAI,AICHI 48003,JAPAN
[2] TOKYO METROPOLITAN UNIV,FAC SCI,DEPT CHEM,HACHIOJI,TOKYO 19203,JAPAN
关键词
D O I
10.1074/jbc.272.47.29934
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
Phosphorylation of alpha B-crystallin, a member of the hsp27 family, in human glioma (U373 MG) cells was stimulated by exposure of the cells to various stimuli, which included heat, arsenite, phorbol 12-myristate 13-acetate (PMA), okadaic acid, H2O2, anisomycin, and high concentrations of NaCl or sorbitol, but not in response to agents that elevated intracellular levels of cyclic AMP. Cells exposed to PMA together with okadaic acid yielded three bands of P-32-labeled alpha B-crystallin when immunoprecipitated samples were subjected to electrophoresis on an isoelectric focusing gel. All of the phosphorylated residues were identified as serine, an indication that three different serine residues can act as sites of phosphorylation in alpha B-crystallin. Structural analysis by mass spectrometry revealed that phosphorylation of alpha B-crystallin occurred at serines 19, 45, and 59. Dithiothreitol and staurosporine selectively inhibited the phosphorylation induced by arsenite and the phorbol ester, respectively, SB202190, an inhibitor of p38 mitogen-activated protein (MAP) kinase, suppressed the phosphorylation induced by arsenite, anisomycin, H2O2, sorbitol, NaCl, and heat shock, but not that induced by PMA and okadaic acid, The PMA-induced phosphorylation was selectively suppressed by an inhibitor of p44 MAP kinase kinase, PD98059, Although PMA and arsenite preferentially stimulated the phosphorylation of Ser-45 and Ser-59, respectively, as determined with antibodies that recognized the respective phosphorylated forms of alpha B-crystallin, all three sites were phosphorylated in response to each stimulus, These results suggest that p38 MAP kinase or p44 MAP kinase might be involved in the signal transduction cascade that leads to the phosphorylation of alpha B-crystallin, The phosphorylation of alpha B-crystallin was also enhanced in the heart and diaphragm when rats were exposed to heat stress (42 degrees C for 20 min).
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页码:29934 / 29941
页数:8
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共 48 条
  • [1] ARRIGO AP, 1987, J BIOL CHEM, V262, P15359
  • [3] ALPHA-B-CRYSTALLIN IS CONSTITUTIVELY EXPRESSED IN CULTURES OF BOVINE ARTICULAR CHONDROCYTES
    BENNARDINI, F
    JULIANO, C
    BENETTI, D
    MIAN, M
    CHIESI, M
    MATTANA, A
    FRANCONI, F
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1995, 208 (02) : 742 - 747
  • [4] BENNDORF R, 1994, J BIOL CHEM, V269, P20780
  • [5] ALPHA-B SUBUNIT OF LENS-SPECIFIC PROTEIN ALPHA-CRYSTALLIN IS PRESENT IN OTHER OCULAR AND NON-OCULAR TISSUES
    BHAT, SP
    NAGINENI, CN
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1989, 158 (01) : 319 - 325
  • [6] THE PHOSPHORYLATION SITES OF THE B-2 CHAIN OF BOVINE ALPHA-CRYSTALLIN
    CHIESA, R
    GAWINOWICZKOLKS, MA
    KLEIMAN, NJ
    SPECTOR, A
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 144 (03) : 1340 - 1347
  • [7] DEFINITION AND COMPARISON OF THE PHOSPHORYLATION SITES OF THE A-CHAIN AND B-CHAIN OF BOVINE ALPHA-CRYSTALLIN
    CHIESA, R
    GAWINOWICZKOLKS, MA
    KLEIMAN, NJ
    SPECTOR, A
    [J]. EXPERIMENTAL EYE RESEARCH, 1988, 46 (02) : 199 - 208
  • [8] HUMAN ALPHA-B-CRYSTALLIN GENE AND PREFERENTIAL PROMOTER FUNCTION IN LENS
    DUBIN, RA
    ALLY, AH
    CHUNG, S
    PIATIGORSKY, J
    [J]. GENOMICS, 1990, 7 (04) : 594 - 601
  • [9] EXPRESSION OF THE MURINE ALPHA-B-CRYSTALLIN GENE IS NOT RESTRICTED TO THE LENS
    DUBIN, RA
    WAWROUSEK, EF
    PIATIGORSKY, J
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (03) : 1083 - 1091
  • [10] A SYNTHETIC INHIBITOR OF THE MITOGEN-ACTIVATED PROTEIN-KINASE CASCADE
    DUDLEY, DT
    PANG, L
    DECKER, SJ
    BRIDGES, AJ
    SALTIEL, AR
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (17) : 7686 - 7689