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rod1 is a seedless target gene of hypoxia-induced mir-210rod1无核目标基因的低氧诱导mir - 210
ROD1 Is a Seedless Target Gene of Hypoxia-Induced miR-
210
1 2 3 1 1
Pasquale Fasanaro , Sveva Romani , Christine Voellenkle , Biagina Maimone , Maurizio C. Capogrossi ,
Fabio Martelli3*
1 Istituto Dermopatico dell’Immacolata IRCCS, Roma, Italy, 2 IRCCS Casa Sollievo della Sofferenza, San Giovanni Rotondo, Italy, 3 IRCCS Policlinico San Donato, Milano, Italy
Abstract
Most metazoan microRNA (miRNA) target sites have perfect pairing to the ‘‘seed’’ sequence, a highly conserved region
centering on miRNA nucleotides 2–7. Thus, complementarity to this region is a necessary requirement for target prediction
algorithms. However, also non-canonical miRNA binding can confer target regulation. Here, we identified a seedless target
of miR-210, a master miRNA of the hypoxic response. We analyzed 20 genes that were inversely correlated to miR-210
expression and did not display any complementarity with miR-210 seed sequence. We validated ROD1 (Regulator of
Differentiation 1, also named PTBP3, Polypyrimidine Tract Binding protein 3) as a miR-210 seedless transcript enriched in
miR-210-containing RNA-induced silencing complexes. ROD1 was not indirectly targeted by a miR-210-induced miRNA.
Conversely, we identified a ‘‘centered’’ miR-210 binding site in ROD1 involving 10 consecutive bases in the central portion of
miR-210. Reporter assays showed that miR-210 inhibited ROD1 by the direct binding to this sequence, demonstrating that
ROD1 is a bona fide seedless target of miR-210. As expected, both ROD1 mRNA and protein were down-modulated upon
hypoxia in a miR-210 dependent manner. ROD1 targeting by miR-210 was biologically significant: the rescue of ROD1
inhibition signif
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