Nucleolin promotes homologous DNA pairing in vitro.
We purified to near homogeneity a previously identified 100 kDa mammalian homologous DNA pairing protein. The purified 100 kDa protein also catalyzed high levels of cell-free homologous DNA recombination activity. This ATP-dependent activity was capable of forming conservative recombinant products between two circular, double-stranded DNA molecules. We were unable to detect any DNA polymerase, DNA ligase, or 5' or 3' exonuclease activity associated with this purified material. The purified 100 kDa protein bound silver nitrate as well as a monoclonal antibody specific for nucleolin. A recombinant protein comprised of the Escherichia coli maltos-ebinding protein fused to the carboxyl-terminal two-thirds of human nucleolin possessed homologous DNA pairing activity. These data indicate that the 100 kDa homologous DNA pairing protein is nucleolin. The observation that nucleolin can carry out homologous DNA strand pairing in vitro raises the prospect that it may function similarly in vivo.
Thyagarajan
B
B
Department of Pharmacology, University of Minnesota Medical School, Minneapolis 55455, USA.
Lundberg
R
R
Rafferty
M
M
Campbell
C
C
D020029:Base Pairing / Q000502:physiology*; D002051:Burkitt Lymphoma; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D002850:Chromatography, Gel; D004273:DNA, Neoplasm / Q000378:metabolism*; D004274:DNA, Recombinant / Q000378:metabolism; D006801:Humans; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000502:physiology; D009697:Nucleolus Organizer Region / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured; D000097796:Nucleolin
D004273:DNA, Neoplasm; D004274:DNA, Recombinant; D009687:Nuclear Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D000097796:Nucleolin