Interaction between the Drosophila CAF-1 and ASF1 chromatin assembly factors.
The assembly of newly synthesized DNA into chromatin is essential for normal growth, development, and differentiation. To gain a better understanding of the assembly of chromatin during DNA synthesis, we identified, cloned, and characterized the 180- and 105-kDa polypeptides of Drosophila chromatin assembly factor 1 (dCAF-1). The purified recombinant p180+p105+p55 dCAF-1 complex is active for DNA replication-coupled chromatin assembly. Furthermore, we have established that the putative 75-kDa polypeptide of dCAF-1 is a C-terminally truncated form of p105 that does not coexist in dCAF-1 complexes containing the p105 subunit. The analysis of native and recombinant dCAF-1 revealed an interaction between dCAF-1 and the Drosophila anti-silencing function 1 (dASF1) component of replication-coupling assembly factor (RCAF). The binding of dASF1 to dCAF-1 is mediated through the p105 subunit of dCAF-1. Consistent with the interaction between dCAF-1 p105 and dASF1 in vitro, we observed that dASF1 and dCAF-1 p105 colocalized in vivo in Drosophila polytene chromosomes. This interaction between dCAF-1 and dASF1 may be a key component of the functional synergy observed between RCAF and dCAF-1 during the assembly of newly synthesized DNA into chromatin.
Tyler
J K
JK
Section of Molecular Biology, University of California, San Diego, La Jolla, California 92093-0347, USA.
Collins
K A
KA
Prasad-Sinha
J
J
Amiott
E
E
Bulger
M
M
Harte
P J
PJ
Kobayashi
R
R
Kadonaga
J T
JT
D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002843:Chromatin / Q000737:chemistry*; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins; D008856:Microscopy, Fluorescence; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010455:Peptides / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D021122:Protein Subunits; D056505:Retinoblastoma-Binding Protein 4; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000235:genetics; D014162:Transfection
C110251:Caf1-55 protein, Drosophila; D018797:Cell Cycle Proteins; D002843:Chromatin; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D010455:Peptides; D021122:Protein Subunits; D056505:Retinoblastoma-Binding Protein 4; C491642:asf1 protein, Drosophila