A matrix/scaffold attachment region binding protein: identification, purification, and mode of binding.
Matrix/scaffold attachment regions (MARs/SARs) partition chromatin into functional loop domains. Here we have identified a chicken protein that selectively binds to MARs from the chicken lysozyme locus and to MARs from Drosophila, mouse, and human genes. This protein, named ARBP (for attachment region binding protein), was purified to homogeneity and shown to bind to MARs in a cooperative fashion. ARBP is an abundant nuclear protein and a component of the internal nuclear network. Deletion mutants indicate that multiple AT-rich sequences, if contained in a minimal approximately 350 bp MAR fragment, can lead to efficient binding of ARBP. Furthermore, dimerization mutants show that, to bind ARBP efficiently, MAR sequences can act synergistically over large distances, apparently with the intervening DNA looping out. The binding characteristics of ARBP to MARs reproduce those of unfractionated matrix preparations, suggesting that ARBP is an important nuclear element for the generation of functional chromatin loops.
von Kries
J P
JP
Physiologisch-Chemisches Institut, Universitäts-Krankenhaus Eppendorf, Hamburg, Federal Republic of Germany.
Buhrmester
H
H
Strätling
W H
WH
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D002843:Chromatin / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004264:DNA Topoisomerases, Type I / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004330:Drosophila; D004912:Erythrocytes / Q000378:metabolism; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007668:Kidney / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D009113:Muramidase / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism; D010057:Oviducts / Q000378:metabolism; D011485:Protein Binding; D015183:Restriction Mapping; D013552:Swine
D002843:Chromatin; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D009113:Muramidase; D004264:DNA Topoisomerases, Type I