Faculty Collaboration Database

Peptidylarginine Deiminase 3 (PAD3) Is Upregulated by Prolactin Stimulation of CID-9 Cells and Expressed in the Lactating Mouse Mammary Gland. PLoS One 2016;11(1):e0147503

Date

01/23/2016

Pubmed ID

26799659

Pubmed Central ID

PMC4723263

DOI

10.1371/journal.pone.0147503

Scopus ID

2-s2.0-84958225594 (requires institutional sign-in at Scopus site)   10 Citations

Abstract

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.

Author List

Li G, Hayward IN, Jenkins BR, Rothfuss HM, Young CH, Nevalainen MT, Muth A, Thompson PR, Navratil AM, Cherrington BD

MESH terms used to index this publication - Major topics in bold

Animals
Cells, Cultured
Female
Hydrolases
Janus Kinase 2
Lactation
Mammary Glands, Animal
Mice
Pregnancy
Protein-Arginine Deiminases
STAT5 Transcription Factor
Signal Transduction
Up-Regulation