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Identification of hemicatenane-specific binding proteins by fractionation of Hela nuclei extracts

Abstract : DNA hemicatenanes (HCs) are four-way junctions in which one strand of a doublestranded helix is catenated with one strand of another double-stranded DNA. Frequently mentioned as DNA replication, recombination and repair intermediates, they have been proposed to participate in the spatial organization of chromosomes and in the regulation of gene expression. To explore potential roles of HCs in genome metabolism, we sought to purify proteins capable of binding specifically HCs by fractionating nuclear extracts from HeLa cells. This approach identified three RNA-binding proteins: the Tudorstaphylococcal nuclease domain 1 (SND1) protein and two proteins from the Drosophila behavior human splicing family, the paraspeckle protein component 1 and the splicing factor proline-and glutamine-rich protein. Since these proteins were partially pure after fractionation, truncated forms of these proteins were expressed in Escherichia coli and purified to near homogeneity. The specificity of their interaction with HCs was reexamined in vitro. The two truncated purified SND1 proteins exhibited specificity for HCs, opening the interesting possibility of a link between the basic transcription machinery and HC structures via SND1.
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Contributor : Emmanuelle Delagoutte Connect in order to contact the contributor
Submitted on : Thursday, November 26, 2020 - 8:27:26 AM
Last modification on : Sunday, June 26, 2022 - 2:58:30 AM


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Oumayma Rouis, Cedric Broussard, Francois Guillonneau, Jean-Baptiste Boulé, Emmanuelle Delagoutte. Identification of hemicatenane-specific binding proteins by fractionation of Hela nuclei extracts. Biochemical Journal, 2020, 477 (2), pp.509-524. ⟨10.1042/BCJ20190855⟩. ⟨mnhn-02371714v2⟩



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