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Mapping and use of a sequence that targets DNA ligase I to sites of DNA replication in vivo.

Cardoso, M. Cristina ; Joseph, C. ; Rahn, H. P. ; Reusch, R. ; Nadal-Ginard, B. ; Leonhardt, H. (1997)
Mapping and use of a sequence that targets DNA ligase I to sites of DNA replication in vivo.
In: The Journal of cell biology, 139 (3)
Artikel, Bibliographie

Kurzbeschreibung (Abstract)

The mammalian nucleus is highly organized, and nuclear processes such as DNA replication occur in discrete nuclear foci, a phenomenon often termed "functional organization" of the nucleus. We describe the identification and characterization of a bipartite targeting sequence (amino acids 1-28 and 111-179) that is necessary and sufficient to direct DNA ligase I to nuclear replication foci during S phase. This targeting sequence is located within the regulatory, NH2-terminal domain of the protein and is dispensable for enzyme activity in vitro but is required in vivo. The targeting domain functions position independently at either the NH2 or the COOH termini of heterologous proteins. We used the targeting sequence of DNA ligase I to visualize replication foci in vivo. Chimeric proteins with DNA ligase I and the green fluorescent protein localized at replication foci in living mammalian cells and thus show that these subnuclear functional domains, previously observed in fixed cells, exist in vivo. The characteristic redistribution of these chimeric proteins makes them unique markers for cell cycle studies to directly monitor entry into S phase in living cells.

Typ des Eintrags: Artikel
Erschienen: 1997
Autor(en): Cardoso, M. Cristina ; Joseph, C. ; Rahn, H. P. ; Reusch, R. ; Nadal-Ginard, B. ; Leonhardt, H.
Art des Eintrags: Bibliographie
Titel: Mapping and use of a sequence that targets DNA ligase I to sites of DNA replication in vivo.
Sprache: Englisch
Publikationsjahr: 1997
Titel der Zeitschrift, Zeitung oder Schriftenreihe: The Journal of cell biology
Jahrgang/Volume einer Zeitschrift: 139
(Heft-)Nummer: 3
URL / URN: http://www.cardoso-lab.org/publications/Cardoso_1997.pdf
Kurzbeschreibung (Abstract):

The mammalian nucleus is highly organized, and nuclear processes such as DNA replication occur in discrete nuclear foci, a phenomenon often termed "functional organization" of the nucleus. We describe the identification and characterization of a bipartite targeting sequence (amino acids 1-28 and 111-179) that is necessary and sufficient to direct DNA ligase I to nuclear replication foci during S phase. This targeting sequence is located within the regulatory, NH2-terminal domain of the protein and is dispensable for enzyme activity in vitro but is required in vivo. The targeting domain functions position independently at either the NH2 or the COOH termini of heterologous proteins. We used the targeting sequence of DNA ligase I to visualize replication foci in vivo. Chimeric proteins with DNA ligase I and the green fluorescent protein localized at replication foci in living mammalian cells and thus show that these subnuclear functional domains, previously observed in fixed cells, exist in vivo. The characteristic redistribution of these chimeric proteins makes them unique markers for cell cycle studies to directly monitor entry into S phase in living cells.

Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie > Cell Biology and Epigenetics
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10 Fachbereich Biologie
Hinterlegungsdatum: 05 Mär 2010 15:44
Letzte Änderung: 13 Nov 2014 08:13
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