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Ongoing DNA synthesis in the rat cerebral cortex is regulated by a proteolytic pathway independent of the proteasome and calpains

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Summary

By using mini-units of tissue and protease inhibitors in short term incubation (0–180 min), we studied the role of proteolysis for ongoing DNA replication in the developing rat cerebral cortex. The protease inhibitors TLCK, TPCK, PMSF, MG-132 and PSI markedly inhibited DNA synthesis. The inhibitory effects were concentration-dependent and of early onset (within 60 min). The most selective proteasome inhibitors lactacystin and clasto-lactacystin-β-lactone as well as the calpain inhibitor I and II had no or minimal effects on DNA synthesis. Only high concentrations of calpain inhibitor I (≥ 250 μM) and calpain inhibitor II (≥ 500 μM) gave a DNA synthesis inhibition. These results suggest that (1) ongoing DNA replication is regulated by proteolysis and (2) the proteolytic pathways involved are neither the proteasome nor the calpains.

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Acknowledgements

This work was supported by grants from the Swedish Research Council, the Karolinska Institute and the Stockholms Läns Landsting.

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Authors and Affiliations

  1. Department of Clinical Neuroscience, Huddinge Division of Neurology, Karolinska Institute, Huddinge University Hospital, 141 86, Huddinge, Sweden

    J. Sebastián Yakisich, Åke Sidén & Mabel Cruz

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  1. J. Sebastián Yakisich
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  2. Åke Sidén
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  3. Mabel Cruz
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Correspondence to J. Sebastián Yakisich.

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Table S1

Proteases inhibited by TLCK, TPCK, PMSF, MG-132, PSI, LC, CLC, Ca I I and Ca I II. A) Reported IC50 for the proteasome and calpains. B) Reported IC50 for the other cysteine and serine proteases (172 kb).

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Yakisich, J.S., Sidén, Å. & Cruz, M. Ongoing DNA synthesis in the rat cerebral cortex is regulated by a proteolytic pathway independent of the proteasome and calpains. Invest New Drugs 28, 242–250 (2010). https://doi.org/10.1007/s10637-009-9238-4

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