Dataset Information

Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome.

ABSTRACT: Citrullination is a post-translational modification (PTM) in which positively charged peptidyl-arginine is converted into neutral peptidyl-citrulline by peptidylarginine deiminase (PAD or PADI) enzymes. The full protein citrullinome in many tissues is unknown. Herein, we used mass spectrometry and identified 107 citrullinated proteins in the lactation day 9 (L9) mouse mammary gland including histone H2A, ?-tubulin, and ?-casein. Given the importance of prolactin to lactation, we next tested if it stimulates PAD-catalyzed citrullination using mouse mammary epithelial CID-9 cells. Stimulation of CID-9 cells with 5 µg/mL prolactin for 10 min induced a 2-fold increase in histone H2A citrullination and a 4.5-fold increase in ?-tubulin citrullination. We next investigated if prolactin-induced citrullination regulates the expression of lactation genes ?-casein (Csn2) and butyrophilin (Btn1a1). Prolactin treatment for 12 h increased ?-casein and butyrophilin mRNA expression; however, this increase was significantly inhibited by the pan-PAD inhibitor, BB-Cl-amidine (BB-ClA). We also examined the effect of tubulin citrullination on the overall polymerization rate of microtubules. Our results show that citrullinated tubulin had a higher maximum overall polymerization rate. Our work suggests that protein citrullination is an important PTM that regulates gene expression and microtubule dynamics in mammary epithelial cells.

SUBMITTER: Li G

PROVIDER: S-EPMC7177251 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome.

Li Guangyuan G Young Coleman H CH Snow Bryce B Christensen Amanda O AO Demoruelle M Kristen MK Nemmara Venkatesh V VV Thompson Paul R PR Rothfuss Heather M HM Cherrington Brian D BD

International journal of molecular sciences 20200410 7

Citrullination is a post-translational modification (PTM) in which positively charged peptidyl-arginine is converted into neutral peptidyl-citrulline by peptidylarginine deiminase (PAD or PADI) enzymes. The full protein citrullinome in many tissues is unknown. Herein, we used mass spectrometry and identified 107 citrullinated proteins in the lactation day 9 (L9) mouse mammary gland including histone H2A, α-tubulin, and β-casein. Given the importance of prolactin to lactation, we next tested if i ...[more]

PMID: 32290104

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Project description:Background: The mammary gland is a dynamic organ that undergoes important physiological changes during reproductive cycles. Until now, data regarding the characterisation of miRNA in the mammary gland have been scarce and mainly focused on their abnormal expression in breast cancer. Our goal was to characterise the microRNA (miRNA) involved in mechanisms regulating the mammary function, with particular focus on the lactation stage Methodology/principal findings: Using high-throughput sequencing technology, an exhaustive repertoire of miRNA (miRNome) expressed in mouse and bovine mammary glands during established lactation was identified, characterised and compared. Furthermore, in order to obtain more information on miRNA loading in the RNA-induced silencing complex (RISC), the miRNome was compared with that obtained from RNA associated with the AGO2 protein (AGO2-miRNome) in mouse lactating mammary gland. This study enabled the identification of 164 and 167 miRNA in mouse and bovine, respectively. Among the 30 miRNA most highly expressed in each species, 24 were common to both species and six of them tended to be more highly expressed in lactating than in non-lactating mammary gland. The potential functional roles of these 24 miRNA were deduced using DIANA-miRPath software, based on miRNA/mRNA interactions. Moreover, seven putative novel miRNA were identified. Using DAVID analysis, it was concluded that the predicted targets of two of these putative novel miRNA are involved in mammary gland morphogenesis. Conclusion/significance: Our study provides an overview of the characteristics of lactating mouse and bovine mammary gland miRNA expression profiles. Moreover, species-conserved miRNA involved in this fundamental biological function were identified. These miRNomes will new be used as references for further studies during which the impact of animal breeding on the miRNA expression will be analysed. microRNA profiles of mammary glands from 2 FVB/N mice at lactation day-12 and mammary biopsies from 4 Holstein dairy cows at mid-lactation, generated by deep sequencing, using Illumina HiSeq 2000.

Dataset Information

Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome.

Publications

Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome.

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