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The Chediak-Higashi Protein Interacts with SNARE Complex and Signal Transduction Proteins

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Abstract

Background

Chediak-Higashi syndrome (CHS) is an inherited immunodeficiency disease characterized by giant lysosomes and impaired leukocyte degranulation. CHS results from mutations in the lysosomal trafficking regulator (LYST) gene, which encodes a 425-kD cytoplasmic protein of unknown function. The goal of this study was to identify proteins that interact with LYST as a first step in understanding how LYST modulates lysosomal exocytosis.

Materials and Methods

Fourteen cDNA fragments, covering the entire coding domain of LYST, were used as baits to screen five human cDNA libraries by a yeast two-hybrid method, modified to allow screening in the activation and the binding domain, three selectable markers, and more stringent confirmation procedures. Five of the interactions were confirmed by an in vitro binding assay.

Results

Twenty-one proteins that interact with LYST were identified in yeast two-hybrid screens. Four interactions, confirmed directly, were with proteins important in vesicular transport and signal transduction (the SNARE-complex protein HRS, 14-3-3, and casein kinase II).

Conclusions

On the basis of protein interactions, LYST appears to function as an adapter protein that may juxtapose proteins that mediate intracellular membrane fusion reactions. The pathologic manifestations observed in CHS patients and in mice with the homologous mutation beige suggest that understanding the role of LYST may be relevant to the treatment of not only CHS but also of diseases such as asthma, urticaria, and lupus, as well as to the molecular dissection of the CHS-associated cancer predisposition.

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Acknowledgments

We thank Naomi Kitamura for provision of the anti-Hrs antibody and Pietro de Camilli for reviewing the manuscript. S.F.K. was supported by the American Cancer Society, the Arthritis Foundation, the Howard Hughes Medical Institute, and the National Institutes of Health.

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Authors and Affiliations

  1. CuraGen Corporation, New Haven, CT, USA

    Velizar T. Tchernev, Traci A. Mansfield, Loic Giot, A. Madan Kumar, Krishnan Nandabalan, Ying Li, Vishnu S. Mishra, Jonathan M. Rothberg & Stephen F. Kingsmore

  2. Departments of Medicine, Pathology and Laboratory Medicine, Pediatrics, and Center for Mammalian Genetics, University of Florida, Gainesville, FL, USA

    Velizar T. Tchernev, Vishnu S. Mishra, John C. Detter, Margaret R. Wallace, Frederick S. Southwick & Stephen F. Kingsmore

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  1. Velizar T. Tchernev
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Correspondence to Velizar T. Tchernev.

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Tchernev, V.T., Mansfield, T.A., Giot, L. et al. The Chediak-Higashi Protein Interacts with SNARE Complex and Signal Transduction Proteins. Mol Med 8, 56–64 (2002). https://doi.org/10.1007/BF03402003

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