Histones in transit: Cytosolic histone complexes and diacetylation of H4 during nucleosome assembly in human cells

被引:137
作者
Chang, L
Loranger, SS
Mizzen, C
Ernst, SG
Allis, CD
Annunziato, AT
机构
[1] BOSTON COLL, DEPT BIOL, CHESTNUT HILL, MA 02167 USA
[2] UNIV ROCHESTER, DEPT BIOL, ROCHESTER, NY 14627 USA
[3] TUFTS UNIV, DEPT BIOL, MEDFORD, MA 02155 USA
关键词
D O I
10.1021/bi962069i
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
The organization and acetylation of nascent histones prior to their stable incorporation into chromatin were examined. Through sedimentation and immunoprecipitation analyses of HeLa cytosolic extracts, two somatic non-nucleosomal histone complexes were detected: one containing nascent H3 and H4, and a second containing H2A (and probably H2B) in association with the nonhistone protein NAP-1. The H3/H4 complex has a sedimentation coefficient of 5-6S, consistent with the presence of one or more escort proteins. H4 in the cytosolic H3/H4 complex is diacetylated, fully in accord with the acetylation state of newly synthesized H4 in chromatin. The diacetylation of nascent human H4 is therefore completed prior to nucleosome assembly. As part of our studies of the nascent H3/H4 complex, the cytoplasmic histone acetyltransferase most likely responsible for acetylating newly synthesized H4 was also investigated. HeLa histone acetyltransferase B (HAT B) acetylates H4 but not H3 in vitro, and maximally diacetylates H4 even in the presence of sodium butyrate. Human HAT B acetylates H4 exclusively on the lysine residues at positions 5 and 12, in complete agreement with the highly conserved acetylation pattern of nascent nucleosomal H4 (Sobel et al., 1995), and has a native molecular weight of similar to 100 kDa. Based on our findings a model is presented for the involvement of histone acetylation and NAP-1 in H2A/H2B deposition and exchange, during nucleosome assembly and chromatin remodeling in vivo.
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页码:469 / 480
页数:12
相关论文
共 90 条
[71]   PURIFICATION AND CHARACTERIZATION OF CAF-I, A HUMAN CELL FACTOR REQUIRED FOR CHROMATIN ASSEMBLY DURING DNA-REPLICATION INVITRO [J].
SMITH, S ;
STILLMAN, B .
CELL, 1989, 58 (01) :15-25
[72]  
SOBEL RE, 1994, J BIOL CHEM, V269, P18576
[73]   CONSERVATION OF DEPOSITION-RELATED ACETYLATION SITES IN NEWLY SYNTHESIZED HISTONES H3 AND H4 [J].
SOBEL, RE ;
COOK, RG ;
PERRY, CA ;
ANNUNZIATO, AT ;
ALLIS, CD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (04) :1237-1241
[74]   GENERATION OF DIFFERENT NUCLEOSOME SPACING PERIODICITIES INVITRO - POSSIBLE ORIGIN OF CELL TYPE SPECIFICITY [J].
STEIN, A ;
MITCHELL, M .
JOURNAL OF MOLECULAR BIOLOGY, 1988, 203 (04) :1029-1043
[75]  
STEIN A, 1980, J BIOL CHEM, V255, P3629
[76]   CHROMATIN ASSEMBLY DURING SV40 DNA-REPLICATION INVITRO [J].
STILLMAN, B .
CELL, 1986, 45 (04) :555-565
[77]   HISTONE-SPECIFIC ACETYLTRANSFERASES FROM CALF THYMUS - ISOLATION, PROPERTIES, AND SUBSTRATE-SPECIFICITY OF 3 DIFFERENT ENZYMES [J].
SURES, I ;
GALLWITZ, D .
BIOCHEMISTRY, 1980, 19 (05) :943-951
[78]   OCTAMER OF HISTONES IN CHROMATIN AND FREE IN SOLUTION [J].
THOMAS, JO ;
KORNBERG, RD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1975, 72 (07) :2626-2630
[79]   HISTONE-H4 ACETYLATION IN HUMAN-CELLS - FREQUENCY OF ACETYLATION AT DIFFERENT SITES DEFINED BY IMMUNOLABELING WITH SITE-SPECIFIC ANTIBODIES [J].
TURNER, BM ;
ONEILL, LP ;
ALLAN, IM .
FEBS LETTERS, 1989, 253 (1-2) :141-145
[80]  
van Holde K.E., 1988, CHROMATIN