Peptidylarginine Deiminase 3 (PAD3) Is Upregulated by Prolactin Stimulation of CID-9 Cells and Expressed in the Lactating Mouse Mammary Gland

Autor: Isaac N. Hayward, Marja T. Nevalainen, Aaron Muth, Guangyuan Li, Heather M. Rothfuss, Amy M. Navratil, Brittany D. Jenkins, Paul R. Thompson, Brian D. Cherrington, Coleman H. Young
Rok vydání: 2016
Předmět:
0301 basic medicine
Physiology
Hydrolases
Peptide Hormones
Maternal Health
Mammary gland
Biochemistry
Protein citrullination
Histones
Mice
Endocrinology
Protein-Arginine Deiminase Type 3
Reproductive Physiology
Pregnancy
Lactation
Medicine and Health Sciences
STAT5 Transcription Factor
Post-Translational Modification
Cells
Cultured
STAT5
Multidisciplinary
Janus kinase 2
biology
Obstetrics and Gynecology
Citrullination
Complementary DNA
Mammary Glands
Up-Regulation
Nucleic acids
medicine.anatomical_structure
Medicine
Female
Anatomy
Signal transduction
Research Article
Signal Transduction
medicine.medical_specialty
Forms of DNA
Science
Research and Analysis Methods
03 medical and health sciences
Exocrine Glands
Mammary Glands
Animal
Internal medicine
DNA-binding proteins
Genetics
medicine
Animals
Immunohistochemistry Techniques
Endocrine Physiology
Reproductive System
Biology and Life Sciences
Proteins
DNA
Janus Kinase 2
Hormones
Prolactin
Histochemistry and Cytochemistry Techniques
030104 developmental biology
Immunologic Techniques
Protein-Arginine Deiminases
biology.protein
Women's Health
Breast Tissue
Zdroj: PLoS ONE, Vol 11, Iss 1, p e0147503 (2016)
PLoS ONE
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0147503
Popis: Peptidylarginine deiminases (PADs) post-translationally convert arginine into neutral citrulline residues. Our past work shows that PADs are expressed in the canine and murine mammary glands; however, the mechanisms regulating PAD expression and the function of citrullination in the normal mammary gland are unclear. Therefore, the first objective herein was to investigate regulation of PAD expression in mammary epithelial cells. We first examined PAD levels in CID-9 cells, which were derived from the mammary gland of mid-pregnant mice. PAD3 expression is significantly higher than all other PAD isoforms and mediates protein citrullination in CID-9 cells. We next hypothesized that prolactin regulates PAD3 expression. To test this, CID-9 cells were stimulated with 5 μg/mL of prolactin for 48 hours which significantly increases PAD3 mRNA and protein expression. Use of a JAK2 inhibitor and a dominant negative (DN)-STAT5 adenovirus indicate that prolactin stimulation of PAD3 expression is mediated by the JAK2/STAT5 signaling pathway in CID-9 cells. In addition, the human PAD3 gene promoter is prolactin responsive in CID-9 cells. Our second objective was to investigate the expression and activity of PAD3 in the lactating mouse mammary gland. PAD3 expression in the mammary gland is highest on lactation day 9 and coincident with citrullinated proteins such as histones. Use of the PAD3 specific inhibitor, Cl4-amidine, indicates that PAD3, in part, can citrullinate proteins in L9 mammary glands. Collectively, our results show that upregulation of PAD3 is mediated by prolactin induction of the JAK2/STAT5 signaling pathway, and that PAD3 appears to citrullinate proteins during lactation.
Databáze: OpenAIRE
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