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Gene Fusion Proteins

The amino terminus targets the mixed lineage leukemia (MLL) protein to the nucleolus, nuclear matrix and mitotic chromosomal scaffolds

Leukemia volume 14, pages 1898–1908 (2000)Cite this article

Abstract

The mixed-lineage leukemia gene (MLL) is associated with more than 25 chromosomal translocations involving band 11q23 in diverse subtypes of human acute leukemia. Conditional expression of a 50 kDa amino terminal fragment spanning the AT hook motifs of MLL (MLL3AT) causes cell cycle arrest, upregulation of p21Cip1 and p27Kip1 and partial monocytic differentiation of the monoblastic U937 cell line, suggesting a major role for MLL3AT in MLL-AF9-induced myelomonocytic differentiation. In this study, we analyzed the subcellular localization of conditionally expressed MLL3AT in both U937 and HeLa cell lines. Immunofluorescence staining, confocal laser scanning microscopy and immunoelectron microscopy indicated that MLL3AT, like endogenous MLL, localized in the nucleoplasm in a punctate pattern of distribution, including regions attached to the nuclear envelope and the periphery of the nucleolus. We found that MLL3AT and endogenous MLL were present in interphase nuclear matrices and colocalized with topoisomerase II to mitotic chromosomal scaffolds. Nucleoplasm and nucleolar localization was observed even for MLL-AF9 and MLL-AF4 conditionally expressed chimeric proteins, suggesting a common target conferred by the amino terminus of MLL to many if not all the chimeric MLL proteins. The nuclear matrix/scaffold association suggests a role for the amino terminus of MLL in the modulation of chromatin structure, leading to epigenetic effects on the maintenance of gene expression.

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Acknowledgements

We wish to thank Drs AT Look and M Introna for helpful discussions. We also thank Dr S Korsmeyer for the gift of anti-MLL 2F7 antibody, Dr JH Kersey for the gift of anti-AF4 C15 antibody, Dr PG Pelicci for MLL cDNA fragments and Dr G Grosveld for tet-regulatable U937T cell line. This work was supported by the National Institutes of Health grant CA21765, the American Lebanese Syrian Associated Charities, and by the Fondazione ‘Matilde Tettamanti’. CC was supported in part by a fellowship from the Associazione Italiana per la Ricerca sul Cancro and the Fondazione Italiana per la Ricerca sul Cancro. ASA was supported in part by the Instituto Nacional de Cancerologia, Bogota’, Colombia.

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Authors and Affiliations

  1. Centro di Ricerca ‘M Tettamanti’, Clinica Pediatrica Università di Milano – Bicocca, Ospedale S Gerardo, Monza, Italy

    C Caslini, ASerna Alarcòn & A Biondi

  2. Department of Experimental Oncology, St Jude Children's Research Hospital, Memphis, TN, USA

    C Caslini & R Tanaka

  3. Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA, USA

    JL Hess

  4. Department of Clinical Oncology – Institute of Medical Science, University of Tokyo, Japan

    R Tanaka

  5. Department of Virology/Molecular Biology, St Jude Children's Research Hospital, Memphis, TN, USA

    KG Murti

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Caslini, C., Alarcòn, A., Hess, J. et al. The amino terminus targets the mixed lineage leukemia (MLL) protein to the nucleolus, nuclear matrix and mitotic chromosomal scaffolds. Leukemia 14, 1898–1908 (2000). https://doi.org/10.1038/sj.leu.2401933

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