The high-mobility group protein T160 binds to both linear and cruciform DNA and mediates DNA bending as determined by ring closure.
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.
Gariglio
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M
Institute of Microbiology, Medical School of Novara, University of Torino, Italy.
Ying
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GG
Hertel
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L
Gaboli
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M
Clerc
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RG
Landolfo
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D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology; D001665:Binding Sites; D003001:Cloning, Molecular; D015342:DNA Probes; D004260:DNA Repair; D004261:DNA Replication; D004270:DNA, Circular / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008811:Mice, Inbred DBA; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011995:Recombination, Genetic; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D004270:DNA, Circular; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; C070454:Ssrp1 protein, mouse