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Originally published In Press as doi:10.1074/jbc.M104730200 on August 9, 2001

J. Biol. Chem., Vol. 276, Issue 42, 39320-39329, October 19, 2001
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Interaction between Pyrin and the Apoptotic Speck Protein (ASC) Modulates ASC-induced Apoptosis*

Neil RichardsDagger §, Philip SchanerDagger §||¶¶, Arturo Diaz**||||, Jeanne StuckeyDagger Dagger , Eric SheldenDagger , Anish WadhwaDagger , and Deborah L. GumucioDagger §§

From the Departments of Dagger  Cell and Developmental Biology, ** Internal Medicine, and Dagger Dagger  Biochemistry and the ¶¶ Graduate Program in Cellular and Molecular Biology, The University of Michigan Medical School, Ann Arbor, Michigan, 48109-0616

Patients with familial Mediterranean fever suffer sporadic inflammatory attacks characterized by fever and intense pain (in joints, abdomen, or chest). Pyrin, the product of the MEFV locus, is a cytosolic protein whose function is unknown. Using pyrin as a "bait" to probe a yeast two-hybrid library made from neutrophil cDNA, we isolated apoptotic speck protein containing a caspase recruitment domain (CARD) (ASC), a proapoptotic protein that induces the formation of large cytosolic "specks" in transfected cells. We found that when HeLa cells are transfected with ASC, specks are formed. After co-transfection of cells with ASC plus wild type pyrin, an increase in speck-positive cells is found, and speck-positive cells show increased survival. Immunofluorescence studies show that pyrin co-localizes with ASC in specks. Speck localization requires exon 1 of pyrin, but exon 1 alone of pyrin does not result in an increase in the number of specks. Exon 1 of pyrin and exon 1 of ASC show 42% sequence similarity and resemble death domain-related structures in modeling studies. These findings link pyrin to apoptosis pathways and suggest that the modulation of cell survival may be a component of the pathophysiology of familial Mediterranean fever.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

Supported by Hematology Training Grant T32-HL07622.

|| Supported by Organogenesis Training Grant T32-HD07505.

§§ Supported by a grant from the Arthritis Foundation. To whom correspondence should be addressed: Dept. of Cell and Developmental Biology, University of Michigan Medical School, 5704 Medical Science II, Ann Arbor, MI 48109-0616. Tel.: 734-647-0172; Fax: 734-647-9559; E-mail: dgumucio@umich.edu.

|||| Supported by Rheumatology Training Grant T32-AR07080.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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