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Cullin 4A and 4B ubiquitin ligases interact with γ-tubulin and induce its polyubiquitination

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Abstract

Regulated polyubiquitination is a key step for controlling protein degradation and maintaining proper balance between the proliferation of normal and uncontrolled cells. Addition of ubiquitin to the proteins by E3 ubiquitin ligases targets them for degradation by the 26S proteosome machinery. Discrepancies in ubiquitination and/or proteosome degradation might lead to multiple genetic disorders in humans. It is reported that CUL1 and BRCA1 ubiquitin ligases localize on centrosome region and regulate the centrosome duplication cycle for genomic stability. In the current study, we predicted the possible interaction of E3 ubiquitin ligase CUL4A complex with γ-tubulin, a centrosome-specific protein, using bioinformatic protein–protein docking analysis. We also confirmed their interaction by performing co-immunoprecipitation studies using endogenous CUL4A/B and stable cell lines that overexpress Flag-CUL4A or Flag-CUL4B. We additionally noted that the γ-tubulin was polyubiquitinated by CUL4A or 4B immune complex indicating that CUL4A or CUL4B may regulate the stability of γ-tubulin. Furthermore, the inhibition of proteosomal degradation pathway using MG132 or LLNV drugs resulted in accumulation and co-localization of CUL4A with γ-tubulin in the centrosome region. Overall, our observation has identified γ-tubulin as a novel target for E3 ubiquitin ligase CUL4 complex, and might lead to the establishment of a unique mechanism for controlling centrosome stability.

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Acknowledgments

We are very grateful to Prof. Pradip Raychaudhuri and Dr. Srilata Bagchi, University of Illinois at Chicago, USA for providing necessary plasmids, technical advice and constant support. We especially thank Drs. Arunkumar Dhayalan and Anil K. Suresh (Ramalingaswamy Fellow) for critical reading of the manuscript. This work is supported by Department of Science and Technology-SERB, INDIA Grants; SR/FT/LS-76/2011and SB/EMEQ-038/2013 to Dr. Sudhakar Baluchamy. Miss. Prachi Singh is a recipient of DBT-JRF. Mr. M. Kannan is a recipient of a Rajiv Gandhi national research fellowship of the UGC (No. F. 14-2(SC)/2009 (SA-III).We apologize for not being able to comprehensively cite references related to CUL4 and centrosome due to focused topics discussed in this manuscript.

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Authors and Affiliations

  1. Stem Cell Laboratory, Department of Biotechnology, Pondicherry Central University, R. V. Nagar, Kalapet, Pondicherry, 605014, India

    Anand Thirunavukarasou, Gokulapriya Govindarajalu, Prachi Singh, Venkateshwarlu Bandi & Sudhakar Baluchamy

  2. Centre for Bioinformatics, Pondicherry Central University, Pondicherry, 605014, India

    Kannan Muthu

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Thirunavukarasou, A., Govindarajalu, G., Singh, P. et al. Cullin 4A and 4B ubiquitin ligases interact with γ-tubulin and induce its polyubiquitination. Mol Cell Biochem 401, 219–228 (2015). https://doi.org/10.1007/s11010-014-2309-7

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