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J Biol Chem, Vol. 274, Issue 37, 26249-26258, September 10, 1999

Cloning and Expression of Bovine Neutrophil -Defensins
BIOSYNTHETIC PROFILE DURING NEUTROPHILIC MATURATION AND LOCALIZATION OF MATURE PEPTIDE TO NOVEL CYTOPLASMIC DENSE GRANULES

Nannette Y. Yount, Jun Yuan, Alan Tarver^§, Tammy Castro^¶, Gill Diamond^¶, Patti A. Tran, Jack N. Levy, Cheryl McCullough, James S. Cullor, Charles L. Bevins^**, and Michael E. Selsted

From the Departments of  Pathology and  Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine, California 92697, the ^§ Division of Genetics and Molecular Biology, Children's Hospital of Philadelphia and Departments of Pediatrics and Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, the ^¶ Department of Anatomy, Cell Biology and Injury Sciences, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103, the  Department of Veterinary Pathology, School of Veterinary Medicine, University of California, Davis, California 95616, and the ^** Department of Immunology, Cleveland Clinic Foundation Research Institute, Cleveland, Ohio 44195

-Defensins are microbicidal peptides implicated in host defense functions of phagocytic leukocytes and certain surface epithelial cells. Here we investigated the genetic structures and cellular expression of BNBD-4, -12, and -13, three prototypic bovine neutrophil -defensins. Characterization of the corresponding cDNAs indicated that BNBD-4 (41 residues) derives from a 63-amino acid prepropeptide and that BNBD-12 (38 residues) and BNBD-13 (42 residues) derive from a common 60-amino acid precursor (BNBD-12/13). The peptides were found to be encoded by two-exon genes that are closely related to bovine epithelial -defensin genes. BNBD-4 and BNBD-12/13 mRNAs were most abundant in bone marrow, but were expressed differentially in certain non-myeloid tissues. In situ hybridization and immunohistochemical studies demonstrated that BNBD-4 synthesis is completed early in myelopoiesis. BNBD-12 was localized exclusively to the novel dense granules, organelles that also contain precursors of cathelicidins, antimicrobial peptides that undergo proteolytic processing during phagocytosis. In contrast to cathelicidins, Western blot analyses revealed that mature -defensins are the predominant organellar form in myeloid cells. Stimulation of neutrophils with phorbol myristate acetate induced secretion of BNBD-12, indicating that it is co-secreted with pro-cathelicidins. The exocytosis of BNBD-12 by activated neutrophils reveals different mobilization pathways for myeloid - and -defensins.

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Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.

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