Mollie E Jacobs1, Lauren A Jeffers1, Amanda K Welch2, Charles S Wingo2, Brian D Cain3. 1. Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610, USA. 2. Department of Nephrology, University of Florida, Gainesville, FL 32610, USA; North Florida/South Georgia Veterans Health System, Gainesville, FL 32610, USA. 3. Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610, USA. Electronic address: bcain@ufl.edu.
Abstract
AIMS: Recently, microRNAs (miRNAs) have been implicated in control of Edn1 mRNA in several tissues. Here we examined the role of miRNA action on Edn1 mRNA expression in renal distal collecting duct cells. MAIN METHODS: A microarray study was conducted to provide a comprehensive assessment of miRNAs present in a murine inner medullary collecting duct (mIMCD-3) cell line. The experiment was designed as a comparison between mIMCD-3 cells grown in the presence and absence of aldosterone. Argonaute (Ago) immunoprecipitation experiments were used to investigate binding of the RNA induced silencing complex (RISC) to Edn1 mRNA. KEY FINDINGS: Thirty-four miRNAs were detected in very high abundance in mIMCD-3 cells, and a large number of others were present at lower levels. The microarray experiments were validated by quantitative PCR analysis of selected miRNAs. The microarray data, in combination with in silico examination of the Edn1 3' UTR provided a panel of candidate miRNAs that could act upon the Edn1 expression. Edn1 mRNA was co-immunoprecipitated with an Argonaute protein antibody, and this interaction was blocked by anti-miR-709 oligonucleotides. SIGNIFICANCE: These results define the miRNA landscape of the mIMCD-3 cell line. Moreover, Edn1 was shown to interact with Argonaute protein suggesting that it is a target of the RNA induced silencing complex (RISC).
AIMS: Recently, microRNAs (miRNAs) have been implicated in control of Edn1 mRNA in several tissues. Here we examined the role of miRNA action on Edn1 mRNA expression in renal distal collecting duct cells. MAIN METHODS: A microarray study was conducted to provide a comprehensive assessment of miRNAs present in a murine inner medullary collecting duct (mIMCD-3) cell line. The experiment was designed as a comparison between mIMCD-3 cells grown in the presence and absence of aldosterone. Argonaute (Ago) immunoprecipitation experiments were used to investigate binding of the RNA induced silencing complex (RISC) to Edn1 mRNA. KEY FINDINGS: Thirty-four miRNAs were detected in very high abundance in mIMCD-3 cells, and a large number of others were present at lower levels. The microarray experiments were validated by quantitative PCR analysis of selected miRNAs. The microarray data, in combination with in silico examination of the Edn1 3' UTR provided a panel of candidate miRNAs that could act upon the Edn1 expression. Edn1 mRNA was co-immunoprecipitated with an Argonaute protein antibody, and this interaction was blocked by anti-miR-709 oligonucleotides. SIGNIFICANCE: These results define the miRNA landscape of the mIMCD-3 cell line. Moreover, Edn1 was shown to interact with Argonaute protein suggesting that it is a target of the RNA induced silencing complex (RISC).
Authors: Francisco M Reimunde; Cristina Castañares; Mariano Redondo-Horcajo; Santiago Lamas; Fernando Rodríguez-Pascual Journal: Biochem J Date: 2005-05-01 Impact factor: 3.857
Authors: Lisa R Stow; Michelle L Gumz; I Jeanette Lynch; Megan M Greenlee; Alicia Rudin; Brian D Cain; Charles S Wingo Journal: J Biol Chem Date: 2009-07-28 Impact factor: 5.157
Authors: Peter Johnström; Tim D Fryer; Hugh K Richards; Neil G Harris; Olivier Barret; John C Clark; John D Pickard; Anthony P Davenport Journal: Br J Pharmacol Date: 2005-01 Impact factor: 8.739
Authors: Sneha Surendran; Victoria N Jideonwo; Chris Merchun; Miwon Ahn; John Murray; Jennifer Ryan; Kenneth W Dunn; Janaiah Kota; Núria Morral Journal: Sci Rep Date: 2016-01-08 Impact factor: 4.379
Authors: Lauren G Douma; Kristen Solocinski; Sarah H Masten; Dominique H Barral; Sarah J Barilovits; Lauren A Jeffers; Kareme D Alder; Ravi Patel; Charles S Wingo; Kevin D Brown; Brian D Cain; Michelle L Gumz Journal: Front Physiol Date: 2020-03-13 Impact factor: 4.566