There is close agreement in molecular weight (32 versus 36 kDa and both activities require a central AU motif and flanking U residues.
Previous efforts to discover inhibitors of HuR/RNA interaction ich will ein bordell eröffnen yielded a low molecular weight inhibitor using confocal fluctuation spectroscopic assay 51, in which short construct was used for screening and there is no secondary screening included.These facts indicate that there is no domain-domain interaction among three RRMs.A distinct screening strategy using the Alpha Screen method has recently been described using HuR expressed in mammalian cell-based system 52, in which full length HuR was used for screening, and emsa as secondary screening was included.First of all, NMR based method has been developed as a high throughput method compatible to conventional HTS 40 and is now used as orthogonal read-out to the biochemical HTS 41,.Shaw and Kamen ( 5 ) were the first to identify a cis element responsible for regulated mRNA degradation.Although we still use FP as primary HTS, our system is unique.Recent studies have shown that monomers such as uuauuuauu can function as destablizing elements albeit at lower efficiency than the complete ARE element( 10 ).Ota., Suzuki., Nishikawa., Otsuki., Sugiyama., Irie., Wakamatsu., Hayashi., Sato., Nagai., Kimura., Makita., Sekine., Obayashi., Nishi., Shibahara., Tanaka., Ishii., Yamamoto., Saito., Kawai., Isono., Nakamura.This hit rate makes an NMR-based secondary assay practical.




They showed that a 51-nucleotide AU-rich element (ARE) from the 3-UTR of factor could increase the degradation rate of any mRNA( 5 ).1 Publication p Manually curated information that is based on statements in scientific articles for which there is no experimental support.Although the sequence alignment indicates that all three RRMs have the same canonical 112324 fold 19, 20, the function of the individual RRM domains is different.The HuR protein described here may facilitate the generation of such an in vitro system and permit the purification of other factors required for ARE-dependent mRNA degradation.The Elav-like RNA-binding proteins (HuD, HuC, and Hel-N1) are also good candidates for trans-acting factors involved in selective mRNA degradation, since they bind to ARE elements with high affinity and selectivity( 23, 24, 25, 26 ).Editor: Yong-Bin Yan, Tsinghua University, china, received: July 13, 2015; Accepted: September 3, 2015; Published: September 21, 2015, copyright: 2015 Wang.This striking effect most likely arises from the inhibition of HuR oligomerization caused by C11 binding to the protein, since there is no domain-domain interaction among three RRMs ( 49, S4 Fig ).The utility of the hsqc spectra in compound screening and characterization may be further improved if individual domain constructs are used to detect interaction with the hit compounds.


[L_RANDNUM-10-999]