title: Growth inhibition by miR-519 via multiple p21-inducing pathways.
abstract: The microRNA miR-519 robustly inhibits cell proliferation, in turn triggering senescence and decreasing tumor growth. However, the molecular mediators of miR-519-elicited growth inhibition are unknown. Here, we systematically investigated the influence of miR-519 on gene expression profiles leading to growth cessation in HeLa human cervical carcinoma cells. By analyzing miR-519-triggered changes in protein and mRNA expression patterns and by identifying mRNAs associated with biotinylated miR-519, we uncovered two prominent subsets of miR-519-regulated mRNAs. One subset of miR-519 target mRNAs encoded DNA maintenance proteins (including DUT1, EXO1, RPA2, and POLE4); miR-519 repressed their expression and increased DNA damage, in turn raising the levels of the cyclin-dependent kinase (cdk) inhibitor p21. The other subset of miR-519 target mRNAs encoded proteins that control intracellular calcium levels (notably, ATP2C1 and ORAI1); their downregulation by miR-519 aberrantly elevated levels of cytosolic [Ca(2+)] storage in HeLa cells, similarly increasing p21 levels in a manner dependent on the Ca(2+)-activated kinases CaMKII and GSK3β. The rises in levels of DNA damage, the Ca(2+) concentration, and p21 levels stimulated an autophagic phenotype in HeLa and other human carcinoma cell lines. As a consequence, ATP levels increased, and the level of activity of the AMP-activated protein kinase (AMPK) declined, further contributing to the elevation in the abundance of p21. Our results indicate that miR-519 promotes DNA damage, alters Ca(2+) homeostasis, and enhances energy production; together, these processes elevate the expression level of p21, promoting growth inhibition and cell survival.
authors.affiliation: Laboratory of Molecular Biology and Immunology, Research Resources Branch, National Institute on Aging, NIH, Baltimore, Maryland, USA. abdelmohsenk@grc.nia.nih.gov
authors.forename: Kotb
authors.initials: K
authors.lastname: Abdelmohsen
authors.forename: Subramanya
authors.initials: S
authors.lastname: Srikantan
authors.forename: Kumiko
authors.initials: K
authors.lastname: Tominaga
authors.forename: Min-Ju
authors.initials: MJ
authors.lastname: Kang
authors.forename: Yael
authors.initials: Y
authors.lastname: Yaniv
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Martindale
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Yang
authors.forename: Sung-Soo
authors.initials: SS
authors.lastname: Park
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Becker
authors.forename: Murugan
authors.initials: M
authors.lastname: Subramanian
authors.forename: Stuart
authors.initials: S
authors.lastname: Maudsley
authors.forename: Ashish
authors.initials: A
authors.lastname: Lal
authors.forename: Myriam
authors.initials: M
authors.lastname: Gorospe
mesh_terms: D001343:Autophagy; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000235:genetics; D020013:Calcium Signaling; D000252:Calcium-Transporting ATPases / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage; D004257:DNA Polymerase II / Q000235:genetics; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000235:genetics; D004734:Energy Metabolism; D005090:Exodeoxyribonucleases / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D008954:Models, Biological; D000071740:ORAI1 Protein; D000075223:Poly-ADP-Ribose Binding Proteins; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051716:Replication Protein A / Q000235:genetics; D015398:Signal Transduction
chemical_list: C493956:CDKN1A protein, human; D015220:Calcium Channels; D050759:Cyclin-Dependent Kinase Inhibitor p21; C546194:MIRN519 microRNA, human; D035683:MicroRNAs; D000071740:ORAI1 Protein; C510746:ORAI1 protein, human; D000075223:Poly-ADP-Ribose Binding Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051716:Replication Protein A; D004257:DNA Polymerase II; C119525:POLE protein, human; C516794:RPA2 protein, human; C114331:EXO1 protein, human; D005090:Exodeoxyribonucleases; D045643:DNA Repair Enzymes; C403624:ATP2C1 protein, human; D000252:Calcium-Transporting ATPases; D002118:Calcium