Interaction between the Drosophila CAF-1 and ASF1 chromatin assembly factors

Mol Cell Biol. 2001 Oct;21(19):6574-84. doi: 10.1128/mcb.21.19.6574-6584.2001.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Cycle Proteins / metabolism*
  • Chromatin / chemistry*
  • Chromatin Assembly Factor-1
  • Chromosomal Proteins, Non-Histone*
  • Cloning, Molecular
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • DNA-Binding Proteins / physiology
  • Drosophila / genetics*
  • Drosophila / metabolism
  • Drosophila Proteins*
  • Microscopy, Fluorescence
  • Molecular Chaperones*
  • Molecular Sequence Data
  • Nuclear Proteins*
  • Peptides / genetics
  • Peptides / metabolism*
  • Peptides / physiology
  • Protein Subunits
  • Retinoblastoma-Binding Protein 4
  • Sequence Homology, Amino Acid
  • Spodoptera / genetics
  • Transfection

Substances

  • Caf1-55 protein, Drosophila
  • Cell Cycle Proteins
  • Chromatin
  • Chromatin Assembly Factor-1
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Drosophila Proteins
  • Molecular Chaperones
  • Nuclear Proteins
  • Peptides
  • Protein Subunits
  • Retinoblastoma-Binding Protein 4
  • asf1 protein, Drosophila

Associated data

  • GENBANK/AF367177
  • GENBANK/AF367178