Translational Repression of the Disintegrin and Metalloprotease ADAM10 by a Stable G-quadruplex Secondary Structure in Its 5′-Untranslated Region

Abstract

Background: Translation of the ␣-secretase ADAM10 is repressed by its 5Ј-untranslated region (5Ј-UTR). Results: A G-rich region in the ADAM10 5Ј-UTR forms a highly stable G-quadruplex secondary structure, which inhibits translation of a luciferase reporter and ADAM10. Conclusion: The G-quadruplex secondary structure is one inhibitory element for ADAM10 translation. Significance: Our findings provide new insights in the translational regulation of ADAM10.

Translational repression of cyclin D3 by a stable G-quadruplex in its 5′ UTR: implications for cell cycle regulation

“…Specifically, in the presence of 100 mM KCl, the folded structure could not be fully unfolded even at 95°C, which is comparable with other G-quadruplex molecules and suggestive of a very stable structure. 45,46,49,51,52 Notably, the addition of LiCl also increased the T m value of CRQ-wt, although less significantly than did NaCl or KCl, which has been reported and can be explained by the reporter experiments. The reporter plasmids were constructed by insertion of the wild-type or G/U-mutated CRQ sequence exactly in front of the CDS of renilla luciferase and transfected into HEK293T and HepG2 cells.…”

“…41,45 Experimental evidence also supports the existence of RNA G-quadruplexes in the 5' UTRs of several mRNAs, including FGF-2, VEGF, NRAS, Zic-1, ESR-1, MT3-MMP, Bcl-2, TRF2, ADAM10, KRAS, and EBAG9. [45][46][47][48][49][50][51][52][53][54][55] However, not all of the predicted PG4 sequences were found to fold into G-quadruplex structures in vitro. 45 Therefore, there is an urgent need to identify bona fide 5' UTR G-quadruplex sequences, especially for functionally important genes, and to determine the roles of these G-quadruplex structures in diverse cellular processes, for example, in the regulation of cell-cycle progression.…”

“…Moreover, the calculated ΔH° and ΔS° values were also comparable to published data of other G-quadruplexes. 49,52,60 These thermal denaturation experiments were all performed with CRQ-wt RNA oligonucleotides at a concentration of 5 μM. When we conducted these experiments using CRQ-wt oligonucleotides over a 10-fold concentration, ranging from 1 μM to 10 μM, the T m value in the presence of 1 mM KCl showed no apparent change ( Table 3), suggesting that CRQ-wt forms an intramolecular quadruplex.…”

Fe 2+ binds iron responsive element-RNA, selectively changing protein-binding affinities and regulating mRNA repression and activation

“…The recent extensions of the IRE-RNA family to include mRNA encoding a cell cycle protein (40), α-hemoglobin chaperone (41) and amyloid precursor protein (11), in addition to the iron metabolic proteins (e.g., ferritin, ferroportin, and mt-aconitase), indicates that IRE-mRNA control of protein synthesis affects a number of metabolic processes in animal cells. As shown for the IRE-mRNA, other metal metabolite-riboregulator interactions, such as those in mRNA quadruplexes (19,20) that control mRNA activity, may also involve RNA binding of multiple, regulatory proteins.…”

“…Less is known about metal interactions with riboregulators in eukaryotic mRNAs, which function in cytoplasmic protein synthesis uncoupled from mRNA synthesis/DNA transcription, a contrast with bacterial mRNA riboswitches. Recently, metal-sensitive quadruplexes have been shown to function as a protein synthesis rate regulator in a human mRNA (ADAM10;19,20). Our earlier studies showed direct binding of a number of nonferrous metal ions to IRE-RNA, such as Mg 2þ (21,22) and Mn 2þ (12) as well as complexes with Cu 1þ (21) and Ru 2þ (23).…”