A cis-acting region in the N-methyl-d-aspartate R1 3'-untranslated region interacts with the novel RNA-binding proteins beta subunit of alpha glucosidase II and annexin A2-effect of chronic ethanol exposure in vivo

作者：Anji Antje; Kumari Meena^*

来源：European Journal of Neuroscience, 2011, 34(8): 1200-1211.

DOI：10.1111/j.1460-9568.2011.07857.x

• cerebral cortex
• cis-acting regions
• fetal cortical neurons
• mouse
• mRNA stability
• trans-acting proteins

A cis-acting region, Delta 4, located in the 3'-untranslated region of N-methyl-d-aspartate R1(NR1) mRNA interacts with several trans-acting proteins present in polysomes purified from fetal cortical neurons. Chronic ethanol exposure of fetal cortical neurons increases Delta 4 RNAprotein interactions. This increased interaction is due to an increase in one of the Delta 4-binding trans-acting proteins identified as beta subunit of alpha glucosidase II (GII beta). In this study, we examined whether ethanol-mediated regulation of NR1 mRNA in vivo is similar to that in vitro and whether Delta 4trans interactions are important for ethanol-mediated NR1 mRNA stability. Our data show that polysomal proteins from adult mouse cerebral cortex (CC) formed a complex with Delta 4 RNA, suggesting the presence of NR1 mRNA-binding trans-acting proteins in CC polysomes. The intensity of the Delta 4 RNAprotein complex was increased with polysomes from chronic ethanol-exposed CC. The Delta 4 RNAprotein complex harbored GII beta and a second trans-acting protein identified as annexin A2 (AnxA2). Ethanol-sensitive GII beta was upregulated by 70% in ethanol-exposed CC. Heparin, a known binding partner of AnxA2, inhibited Delta 4 RNAprotein complex formation. Transient transfection studies using chimeric constructs with and without the Delta 4 region revealed that cistrans interactions are important for ethanol-mediated stability of NR1 mRNA. Furthermore, our data highlight, for the first time, the presence of a binding site on the 3'-untranslated region of NR1 mRNA for AnxA2 and demonstrate the regulation of NR1 mRNA by AnxA2, GII beta and a third NR1 mRNA-binding protein, which is yet to be identified.

• 出版日期2011-10