• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Nov 1, 1995; 14(21): 5379–5386.
PMCID: PMC394647

The N-terminal domain of human DNA ligase I contains the nuclear localization signal and directs the enzyme to sites of DNA replication.


DNA replication in mammalian cells occurs in discrete nuclear foci called 'replication factories'. Here we show that DNA ligase I, the main DNA ligase activity in proliferating cells, associates with the factories during S phase but displays a diffuse nucleoplasmic distribution in non-S phase nuclei. Immunolocalization analysis of both chloramphenicol acetyltransferase (CAT)-DNA ligase I fusion proteins and epitope tagged DNA ligase I mutants allowed the identification of a 13 amino acid functional nuclear localization signal (NLS) located in the N-terminal regulatory domain of the protein. Furthermore, the NLS is immediately preceded by a 115 amino acid region required for the association of the enzyme with the replication factories. We propose that in vivo the activity of DNA ligase I could be modulated through the control of its sub-nuclear compartmentalization.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barnes DE, Johnston LH, Kodama K, Tomkinson AE, Lasko DD, Lindahl T. Human DNA ligase I cDNA: cloning and functional expression in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6679–6683. [PMC free article] [PubMed]
  • Barnes DE, Tomkinson AE, Lehmann AR, Webster AD, Lindahl T. Mutations in the DNA ligase I gene of an individual with immunodeficiencies and cellular hypersensitivity to DNA-damaging agents. Cell. 1992 May 1;69(3):495–503. [PubMed]
  • Bestor TH. Activation of mammalian DNA methyltransferase by cleavage of a Zn binding regulatory domain. EMBO J. 1992 Jul;11(7):2611–2617. [PMC free article] [PubMed]
  • Bravo R, Macdonald-Bravo H. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol. 1987 Oct;105(4):1549–1554. [PMC free article] [PubMed]
  • Cardoso MC, Leonhardt H, Nadal-Ginard B. Reversal of terminal differentiation and control of DNA replication: cyclin A and Cdk2 specifically localize at subnuclear sites of DNA replication. Cell. 1993 Sep 24;74(6):979–992. [PubMed]
  • Li C, Goodchild J, Baril EF. DNA ligase I is associated with the 21 S complex of enzymes for DNA synthesis in HeLa cells. Nucleic Acids Res. 1994 Feb 25;22(4):632–638. [PMC free article] [PubMed]
  • Dingwall C, Laskey RA. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. [PubMed]
  • Diviacco S, Norio P, Zentilin L, Menzo S, Clementi M, Biamonti G, Riva S, Falaschi A, Giacca M. A novel procedure for quantitative polymerase chain reaction by coamplification of competitive templates. Gene. 1992 Dec 15;122(2):313–320. [PubMed]
  • Elder RH, Rossignol JM. DNA ligases from rat liver. Purification and partial characterization of two molecular forms. Biochemistry. 1990 Jun 26;29(25):6009–6017. [PubMed]
  • Elder RH, Montecucco A, Ciarrocchi G, Rossignol JM. Rat liver DNA ligases. Catalytic properties of a novel form of DNA ligase. Eur J Biochem. 1992 Jan 15;203(1-2):53–58. [PubMed]
  • Hozák P, Hassan AB, Jackson DA, Cook PR. Visualization of replication factories attached to nucleoskeleton. Cell. 1993 Apr 23;73(2):361–373. [PubMed]
  • Hozák P, Jackson DA, Cook PR. Replication factories and nuclear bodies: the ultrastructural characterization of replication sites during the cell cycle. J Cell Sci. 1994 Aug;107(Pt 8):2191–2202. [PubMed]
  • Kemp BE, Pearson RB. Protein kinase recognition sequence motifs. Trends Biochem Sci. 1990 Sep;15(9):342–346. [PubMed]
  • Kodama K, Barnes DE, Lindahl T. In vitro mutagenesis and functional expression in Escherichia coli of a cDNA encoding the catalytic domain of human DNA ligase I. Nucleic Acids Res. 1991 Nov 25;19(22):6093–6099. [PMC free article] [PubMed]
  • Lasko DD, Tomkinson AE, Lindahl T. Mammalian DNA ligases. Biosynthesis and intracellular localization of DNA ligase I. J Biol Chem. 1990 Jul 25;265(21):12618–12622. [PubMed]
  • Lehmann AR, Willis AE, Broughton BC, James MR, Steingrimsdottir H, Harcourt SA, Arlett CF, Lindahl T. Relation between the human fibroblast strain 46BR and cell lines representative of Bloom's syndrome. Cancer Res. 1988 Nov 15;48(22):6343–6347. [PubMed]
  • Leonhardt H, Page AW, Weier HU, Bestor TH. A targeting sequence directs DNA methyltransferase to sites of DNA replication in mammalian nuclei. Cell. 1992 Nov 27;71(5):865–873. [PubMed]
  • Lindahl T, Barnes DE. Mammalian DNA ligases. Annu Rev Biochem. 1992;61:251–281. [PubMed]
  • Montecucco A, Biamonti G, Savini E, Focher F, Spadari S, Ciarrocchi G. DNA ligase I gene expression during differentiation and cell proliferation. Nucleic Acids Res. 1992 Dec 11;20(23):6209–6214. [PMC free article] [PubMed]
  • Montecucco A, Savini E, Biamonti G, Stefanini M, Focher F, Ciarrocchi G. Late induction of human DNA ligase I after UV-C irradiation. Nucleic Acids Res. 1995 Mar 25;23(6):962–966. [PMC free article] [PubMed]
  • Morris GF, Mathews MB. Regulation of proliferating cell nuclear antigen during the cell cycle. J Biol Chem. 1989 Aug 15;264(23):13856–13864. [PubMed]
  • Pedrali-Noy G, Spadari S, Miller-Faurès A, Miller AO, Kruppa J, Koch G. Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin. Nucleic Acids Res. 1980 Jan 25;8(2):377–387. [PMC free article] [PubMed]
  • Prigent C, Lasko DD, Kodama K, Woodgett JR, Lindahl T. Activation of mammalian DNA ligase I through phosphorylation by casein kinase II. EMBO J. 1992 Aug;11(8):2925–2933. [PMC free article] [PubMed]
  • Prigent C, Satoh MS, Daly G, Barnes DE, Lindahl T. Aberrant DNA repair and DNA replication due to an inherited enzymatic defect in human DNA ligase I. Mol Cell Biol. 1994 Jan;14(1):310–317. [PMC free article] [PubMed]
  • Savini E, Biamonti G, Ciarrocchi G, Montecucco A. Cloning and sequence analysis of a cDNA coding for the murine DNA ligase I enzyme. Gene. 1994 Jul 8;144(2):253–257. [PubMed]
  • Tomkinson AE, Lasko DD, Daly G, Lindahl T. Mammalian DNA ligases. Catalytic domain and size of DNA ligase I. J Biol Chem. 1990 Jul 25;265(21):12611–12617. [PubMed]
  • Tomkinson AE, Roberts E, Daly G, Totty NF, Lindahl T. Three distinct DNA ligases in mammalian cells. J Biol Chem. 1991 Nov 15;266(32):21728–21735. [PubMed]
  • Waga S, Bauer G, Stillman B. Reconstitution of complete SV40 DNA replication with purified replication factors. J Biol Chem. 1994 Apr 8;269(14):10923–10934. [PubMed]
  • Wahl AF, Geis AM, Spain BH, Wong SW, Korn D, Wang TS. Gene expression of human DNA polymerase alpha during cell proliferation and the cell cycle. Mol Cell Biol. 1988 Nov;8(11):5016–5025. [PMC free article] [PubMed]
  • Weighardt F, Biamonti G, Riva S. Nucleo-cytoplasmic distribution of human hnRNP proteins: a search for the targeting domains in hnRNP A1. J Cell Sci. 1995 Feb;108(Pt 2):545–555. [PubMed]
  • Zacksenhaus E, Bremner R, Phillips RA, Gallie BL. A bipartite nuclear localization signal in the retinoblastoma gene product and its importance for biological activity. Mol Cell Biol. 1993 Aug;13(8):4588–4599. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...