Evrim BAYRAK KÖMÜRCÜ, Bilge SELÇUK ÖZSAİT, Nihan ÜNALTUNA ERGİNER

Higher expression level of Bat3 is associated with silencing of the Midn gene in primary mouse cardiomyocytes

Midnolin (Midn) is a developmentally regulated nucleolar protein that was originally identified in the mesencephalon of developing mouse embryos. Additionally, Midn expression was detected in various adult tissues and was associated with glucokinase activity in pancreatic beta cells and with the regulation of folliculogenesis in ovaries. In this study, our aim was to analyze the expression and function of the Midn gene in cardiomyocyte cells. We constructed a primary neonatal mouse cardiomyocyte cell culture and analyzed the expression of possible Midn-associated genes after short interfering RNA (siRNA)-mediated gene silencing for Midn. After 24 h of silencing, we observed that the expression level of Bat3 was significantly increased (1.69-fold, P = 0.04). As a result, we have detected a suggestive gene that might act in the common cellular pathways with Midn in cardiomyocyte cells.

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Andres AM, Stotland A, Queliconi BB, Gottlieb RA (2015). A time to reap, a time to sow: mitophagy and biogenesis in cardiac pathophysiology. J Mol Cell Cardiol 78: 62-72.
Calore M, De Windt LJ, Rampazzo A (2015). Genetics meets epigenetics: genetic variants that modulate noncoding RNA in cardiovascular diseases. J Mol Cell Cardiol 89: 27-34.
Chairatvit K, Ngamkitidechakul C (2007). Control of cell proliferation via elevated NEDD8 conjugation in oral squamous cell carcinoma. Mol Cell Biochem 306: 163-169.
Desmots F, Russell HR, Lee Y, Boyd K, McKinnon PJ (2005). The reaper-binding protein scythe modulates apoptosis and proliferation during mammalian development. Mol Cell Biol 25: 10329-10337.
Desmots F, Russell HR, Michel D, McKinnon PJ (2008). Scythe regulates apoptosis-inducing factor stability during endoplasmic reticulum stress-induced apoptosis. J Biol Chem 283: 3264-3271.
Diatchenko L, Lau YF, Campbell AP, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov ED et al. (1996). Suppression subtractive hybridization: a method for generating differentially regulated or tissue specific cDNA probes and libraries. P Natl Acad Sci USA 93: 6025-6030.
Di Tommaso P, Moretti S, Xenarios I, Orobitg M, Montanyola A, Chang JM, Taly JF, Notredame C (2011). T-Coffee: a web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension. Nucleic Acids Res 39: W13-17.
Elia L, Condorelli G (2015). RNA (Epi)genetics in cardiovascular diseases. J Mol Cell Cardiol 89: 11-16.
Gong L, Yeh ET (1999). Identification of the activating and conjugating enzymes of the NEDD8 conjugation pathway. J Biol Chem 274: 12036-12042.
Goto K, Tong KI, Ikura J, Okada H (2011). HLA-B-associated transcript 3 (Bat3/Scythe) negatively regulates Smad phosphorylation in BMP signaling. Cell Death Dis 2: e236.
Grabbe C, Dikic I (2009) Functional roles of ubiquitin-like domain (ULD) and ubiquitin-binding domain (UBD) containing proteins. Chem Rev 109: 1481-1494.
Hofmeister-Brix A, Kollmann K, Langer S, Schultz J, Lenzen S, Baltrusch S (2013). Identification of the ubiquitin-like domain of midnolin as a new glucokinase interaction partner. J Biol Chem 288: 35824-35839.
Kamitani T, Kito K, Nguyen HP, Yeh ET (1997). Characterization of NEDD8, a developmentally down-regulated ubiquitin-like protein. J Biol Chem 272: 28557-28562.
Kandala S, Kim I, Su H (2014). Neddylation and deneddylation in cardiac biology. Am J Cardiovasc Dis 4: 140-158.
Komurcu-Bayrak E, Ozsait B, Erginel-Unaltuna N (2012). Isolation and analysis of genes mainly expressed in adult mouse heart using subtractive hybridization cDNA library. Mol Biol Rep 39: 8065-8074.
Leznicki P, Clancy A, Schwappach B, High S (2010). Bat3 promotes the membrane integration of tail-anchored proteins. J Cell Sci 123: 2170-2178.
Mori F, Nishie M, Piao YS, Kito K, Kamitani T, Takahashi H, Wakabayashi K (2005). Accumulation of NEDD8 in neuronal and glial inclusions of neurodegenerative disorders. Neuropathol Appl Neurobiol 31: 53-61.
Nuss HB, Marban E (1994). Electrophysiological properties of neonatal mouse cardiac myocytes in primary culture. J Physiol 479: 265-279.
Özsait Selçuk B (2003). Analyses of the Subtractive Hybridization Library isolated genes that have a possible role in cardiogenesis. MSc, İstanbul University, İstanbul, Turkey.
Özsait Selçuk B (2010). Heart specific novel genes isolated from the Subtractive Hybridization cDNA Library. PhD, İstanbul University, İstanbul, Turkey.
Rabut G, Peter M (2008). Function and regulation of protein neddylation. Protein modifications: beyond the usual suspects review series. EMBO Rep 9: 969-976.
Ryu KY, Maehr R, Gilchrist CA, Long MA, Bouley DM, Mueller B, Ploegh HL, Kopito RR (2007). The mouse polyubiquitin gene UbC is essential for fetal liver development, cell-cycle progression and stress tolerance. EMBO J 26: 2693-2706.
Ryu KY, Sinnar SA, Reinholdt LG, Vaccari S, Hall S, Garcia MA, Zaitseva TS, Bouley DM, Boekelheide K, Handel MA et al. (2008). The mouse polyubiquitin gene Ubb is essential for meiotic progression. Mol Cell Biol 28: 1136-1146.
Salameh S, Dhein S (2005). Culture of neonatal cardiomyocytes. In: Dhein S, Mohr FW, Delmar M, editors. Practical Methods in Cardiovascular Research. Berlin, Germany: Springer, pp. 568-576.
Sasaki T, Gan EC, Wakeham A, Kornbluth S, Mak TW, Okada H (2007). HLA-B associated transcript 3 (Bat3)/Scythe is essential for p300-mediated acetylation of p53. Genes Dev 21: 848-861.
Schlüter K, Piper HM (2005). Isolation and culture of adult ventricular cardiomyocytes. In: Dhein S, Mohr FW, Delmar M, editors. Practical Methods in Cardiovascular Research. Berlin, Germany: Springer, pp. 557-567.
Schneider CA, Rasband WS, Eliceiri KW (2012). NIH Image to ImageJ: 25 years of image analysis. Nature Methods 9: 671-675.
Sreejit P, Kumar S, Verma RS (2008). An improved protocol for primary culture of cardiomyocyte from neonatal mice. In Vitro Cell Dev Biol-Animal 44: 45-50.
Su H, Li J, Menon S, Liu J, Kumarapeli AR, Wei N, Wang X (2011). Perturbation of cullin deneddylation via conditional Csn8 ablation impairs the ubiquitin-proteasome system and causes cardiomyocyte necrosis and dilated cardiomyopathy in mice. Circ Res 108: 40-50.
Su V, Lau AF (2009). Ubiquitin-like and ubiquitin-associated domain proteins: significance in proteasomal degradation. Cell Mol Life Sci 66: 2819-2833.
Tsukahara M, Suemori H, Noguchi S, Ji ZS, Tsunoo H (2000). Novel nucleolar protein, midnolin, is expressed in the mesencephalon during mouse development. Gene 254: 45-55.
Zielak AE, Canty MJ, Forde N, Coussens PM, Smith GW, Lonergan P, Ireland JJ, Evans AC (2008). Differential expression of genes for transcription factors in theca and granulosa cells following selection of a dominant follicle in cattle. Mol Reprod Dev 75: 904-914