14-3-3 isoforms bind directly Exon B of the 5'UTR of XXXX @entity11028 mRNA.

@entity1 @entity11028 @entity3231
UNASSIGNED: @entity1 @entity3231 , an innate immunity molecule, is encoded by two genes @entity3231 and @entity11028 . The 5' untranslated splice variant of @entity11028 (ABD), but not of @entity3231 (AD), contains exon B (eB). eB is an enhancer for transcription and translation, and contains cis regulatory elements. Specific trans-acting factors, including 14-3-3, bind eB. The 14-3-3 protein family contains 7 isoforms that were found by mass spectrometry to be present in eB electromobility shift assays (REMSA) (AJPLC MP 304:L722, 2013). We investigated whether 14-3-3 isoforms bind directly to eB using four different approaches. i) eB RNA pull down assays showed that 14-3-3 isoforms bind specifically eB. ii) REMSA complexes were formed using purified 14-3-3 isoforms (b, y,   ,   ,   ,   ) with wild type eB RNA, but not with isoform   . Both iii) RNA affinity chromatography assays and vi) Surface Plasmon Resonance analysis showed that 14-3-3 isoforms b, y,   ,   ,   , and   , but not   , bind specifically and directly eB. Inibition of 14-3-3 isoforms y,   ,   , and   /   with shRNAs in @entity1 cells resulted in downregulation of @entity11028 levels, leaving the @entity3231 levels unaffected. However, inhibition of 14-3-3    isoform was correlated with lower levels of both @entity3231 and @entity11028 . Inhibition of 14-3-3   /   isoform that does not bind eB had no effect on the expression levels of @entity3231 and @entity11028 . In conclusion the 14-3-3 protein family affects differential regulation of @entity3231 and @entity11028 by binding directly to @entity11028 5'UTR mRNA.
