Inner nuclear membrane protein LBR preferentially interacts with DNA secondary structures and nucleosomal linker.
The lamin B receptor (LBR) is an integral protein of inner nuclear membrane whose nucleoplasmic amino-terminal domain contributes to the attachment of the membrane to chromatin. Here we analyzed the interactions of a recombinant GST protein containing the amino-terminal domain of the protein with in vitro reconstituted nucleosomes and short DNA fragments. Data show that the LBR amino-terminal domain (AT) binds linker DNA but does not interact with the nucleosome core. Titration and competition studies revealed that the interaction between LBR AT and DNA is saturable, of high affinity (K(D) approximately 4 nM), independent of DNA sequence, and enhanced by DNA curvature and supercoiling. In this respect, LBR amino-terminal domain binding to nucleosomes is similar to that of histone H1 and non histone proteins HMG1/2 which both bind preferentially to linker DNA and present a significant affinity for DNA secondary structures.
Duband-Goulet
I
I
Département de Biologie Cellulaire Institut Jacques Monod, CNRS, Universités Paris VII-Paris VI, Paris, France. dubandg@ijm.jussieu.fr
Courvalin
J C
JC
D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D004276:DNA, Satellite / Q000737:chemistry / Q000378:metabolism; D005982:Glutathione Transferase; D006801:Humans; D007700:Kinetics; D009685:Nuclear Envelope / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D000097682:Lamin B Receptor
D004247:DNA; D004276:DNA, Satellite; D005982:Glutathione Transferase; D009707:Nucleosomes; D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D000097682:Lamin B Receptor