Mapping and Use of a Sequence that Targets DNA Ligase I to Sites of DNA Replication In Vivo
| Autor: | M. Cristina Cardoso, Regina Reusch, Bernardo Nadal-Ginard, Hans-Peter Rahn, Heinrich Leonhardt, Cuthbert Joseph |
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| Jazyk: | angličtina |
| Rok vydání: | 1997 |
| Předmět: |
DNA Replication
DNA Ligases Recombinant Fusion Proteins Green Fluorescent Proteins Eukaryotic DNA replication Biology Peptide Mapping Article DNA replication factor CDT1 DNA Ligase ATP Replication factor C Control of chromosome duplication Animals Humans Amino Acid Sequence Sequence Deletion chemistry.chemical_classification Cell Nucleus DNA ligase Ter protein Cell Cycle DNA replication Cell Biology Peptide Fragments Luminescent Proteins chemistry Biochemistry COS Cells biology.protein Origin recognition complex HeLa Cells |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 1540-8140 0021-9525 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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