The high-mobility group protein T160 binds to both linear and cruciform DNA and mediates DNA bending as determined by ring closure
| Autor: | Guo Guang Ying, Marisa Gariglio, Mirella Gaboli, Santo Landolfo, Roger G. Clerc, Laura Hertel |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
DNA Replication
HMG-box DNA Repair Biology DDB1 Mice SeqA protein domain Tumor Cells Cultured Animals Tissue Distribution Cloning Molecular Replication protein A Recombination Genetic B-Lymphocytes Binding Sites Protein footprinting Ter protein High Mobility Group Proteins Cell Biology DNA-binding domain DNA-Binding Proteins Biochemistry Mice Inbred DBA Biophysics Nucleic Acid Conformation DNA Circular DNA Probes In vitro recombination Protein Binding |
| Zdroj: | Experimental cell research. 236(2) |
| ISSN: | 0014-4827 |
| Popis: | The high-mobility group protein T160 was isolated by screening a phage library from a murine pre-B-cell line L1210. South-Western experiments have previously shown that this protein binds to V-(D)-J recombination signal sequences, suggesting that it may be a sequence-specific DNA-binding protein. However, neither gel-shift nor footprinting analyses have been successfully employed with the T160 protein, despite an extensive effort. In this study, the T160 protein or truncated forms made soluble through denaturing and renaturing cycles in urea were successfully used in gel-shift experiments showing that T160 binds to cruci-form or linear duplex DNA with no apparent sequence specificity. Furthermore, fragments longer than 100 bp efficiently formed covalently closed circular monomers in the presence of T160 and T4 DNA ligase, indicating that the protein is capable of introducing bends into the duplex. Last, tissue distribution by Western blotting analysis showed that the T160 protein is expressed in various murine tissues in addition to those of lymphoid origin. Considering its broad evolutionary conservation (from plants to mammals) also, these results suggest that the functional role of the T160 protein is not limited to V-(D)-J recombination, but might be involved in basic processes such as DNA replication and repairing, where irregular DNA structures are generated and very likely recognized by HMG domain proteins. |
| Databáze: | OpenAIRE |
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