Interactions of the G quartet forming semaphorin 3F RNA with the RGG box domain of the fragile X protein family

Menon, Lakshmi; Mihailescu, Mihaela‐Rita

doi:10.1093/nar/gkm581

Cited by 66 publications

(77 citation statements)

References 49 publications

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“…While the region N-terminal to the common DEAD-box motifs is highly variable in the sequences of Vas orthologs from different species, the presence of RGG repeats within that region is conserved; for example, zebrafish Vas contains nine such motifs, and human Vas (DDX4) contains four. Two RGG repeats are present in mammalian fragile X mental retardation protein (FMRP), and have been shown to specifically recognize a G quartet structure in semaphorin 3F RNA, indicating that this motif can discriminate among target RNAs (Menon and Mihailescu 2007). RGG repeats are often present in proteins that contain other RNA-binding domains; for example, Vas contains a DEAD-box signature, while FMRP contains two hnRNPK homology (KH) domains; thus it has been proposed that they serve an auxiliary role in RNA binding (McBride and Silver 2001).…”

Section: Discussionmentioning

confidence: 99%

Vasa promotes Drosophila germline stem cell differentiation by activating mei-P26 translation by directly interacting with a (U)-rich motif in its 3′ UTR

Liu

Han²,

Lasko³

2009

Genes Dev.

106

105

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Vasa (Vas) is a DEAD-box RNA-binding protein required in Drosophila at several steps of oogenesis and for primordial germ cell (PGC) specification. Vas associates with eukaryotic initiation factor 5B (eIF5B), and this interaction has been implicated in translational activation of gurken mRNA in the oocyte. Vas is expressed in all ovarian germline cells, and aspects of the vas-null phenotype suggest a function in regulating the balance between germline stem cells (GSCs) and their fate-restricted descendants. We used a biochemical approach to recover Vas-associated mRNAs and obtained mei-P26, whose product represses microRNA activity and promotes GSC differentiation. We found that vas and mei-P26 mutants interact, and that mei-P26 translation is substantially reduced in vas mutant cells. In vitro, Vas protein bound specifically to a (U)-rich motif in the mei-P26 39 untranslated region (UTR), and Vas-dependent regulation of GFP-mei-P26 transgenes in vivo was dependent on the same (U)-rich 39 UTR domain. The ability of Vas to activate mei-P26 expression in vivo was abrogated by a mutation that greatly reduces its interaction with eIF5B. Taken together, our data support the conclusion that Vas promotes germ cell differentiation by directly activating mei-P26 translation in early-stage committed cells.[Keywords: Oogenesis; RNA-binding protein; DEAD-box; patterning; development] Supplemental material is available at http://www.genesdev.org.

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Section: Discussionmentioning

confidence: 99%

Vasa promotes Drosophila germline stem cell differentiation by activating mei-P26 translation by directly interacting with a (U)-rich motif in its 3′ UTR

Liu

Han²,

Lasko³

2009

Genes Dev.

106

105

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“…Naturally occurring G-quadruplexes targeted by FMRP adopt multiple conformations and are challenging to study by structural methods (27,28). Therefore, detailed structural information about RGG recognition of G-quadruplexes has been mostly obtained from studies on sc1 RNA (21), the RNA selected against the entire human FMRP but shown to require only the RGG motif of the protein for high affinity and specific binding (29,37,38).…”

Section: Significancementioning

confidence: 99%

“…Some of these RNAs harbor motifs that may form G-quartets and G-quadruplexes in vivo (21, 25). Several mRNAs contain regions shown to form G-quadruplexes implicated in FMRP binding in vitro (21,24,(26)(27)(28)(29). Notably, binding of FMRP to G-rich RNAs in vitro requires only the RGG motif, which specifically interacts with natural and in vitro selected G-quadruplex-containing RNAs such as a 35-nucleotide sc1 RNA (21,(26)(27)(28)(29).…”

mentioning

confidence: 99%

“…Several mRNAs contain regions shown to form G-quadruplexes implicated in FMRP binding in vitro (21,24,(26)(27)(28)(29). Notably, binding of FMRP to G-rich RNAs in vitro requires only the RGG motif, which specifically interacts with natural and in vitro selected G-quadruplex-containing RNAs such as a 35-nucleotide sc1 RNA (21,(26)(27)(28)(29). Recent studies showed that G-quadruplexes facilitate mRNA interactions with ribosome-bound FMRP (30), whereas the RGG motif, in addition to mRNA binding, contributes to association with ribosomes and proteins and translational control (14,(31)(32)(33)(34).…”

mentioning

confidence: 99%

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Crystal structure reveals specific recognition of a G-quadruplex RNA by a β-turn in the RGG motif of FMRP

Vasilyev

Polonskaia

Darnell

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

180

181

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Fragile X Mental Retardation Protein (FMRP) is a regulatory RNA binding protein that plays a central role in the development of several human disorders including Fragile X Syndrome (FXS) and autism. FMRP uses an arginine-glycine-rich (RGG) motif for specific interactions with guanine (G)-quadruplexes, mRNA elements implicated in the disease-associated regulation of specific mRNAs. Here we report the 2.8-Å crystal structure of the complex between the human FMRP RGG peptide bound to the in vitro selected G-rich RNA. In this model system, the RNA adopts an intramolecular K + -stabilized G-quadruplex structure composed of three G-quartets and a mixed tetrad connected to an RNA duplex. The RGG peptide specifically binds to the duplex-quadruplex junction, the mixed tetrad, and the duplex region of the RNA through shape complementarity, cation-π interactions, and multiple hydrogen bonds. Many of these interactions critically depend on a type I β-turn, a secondary structure element whose formation was not previously recognized in the RGG motif of FMRP. RNA mutagenesis and footprinting experiments indicate that interactions of the peptide with the duplex-quadruplex junction and the duplex of RNA are equally important for affinity and specificity of the RGG-RNA complex formation. These results suggest that specific binding of cellular RNAs by FMRP may involve hydrogen bonding with RNA duplexes and that RNA duplex recognition can be a characteristic RNA binding feature for RGG motifs in other proteins.R NA-binding proteins (RBPs) control all aspects of RNA metabolism and are fundamental to core cellular processes. ∼14% of identified human RBPs are implicated in a broad spectrum of human pathologies, including neurodegenerative and muscular diseases, metabolic disorders, and cancers (1, 2). Fragile X Mental Retardation Protein (FMRP) is among the most important RBPs because of its central role in several human diseases (3). Loss of FMRP function due to CGG triplet repeat expansion-associated transcriptional silencing or missense mutations in the protein (4, 5) lead to fragile X syndrome (FXS), the most common cause of inherited intellectual disability. Mutations in FMRP are also the leading monogenic cause of autism (6, 7). Intermediate length repeat expansions in the FMR1 gene are linked to fragile X-associated tremor ataxia syndrome (8) and fragile X-associated primary ovarian insufficiency (9).FMRP contains four canonical nucleic acid-binding motifs, three KH domains and one arginine-glycine-rich (RGG) box, which mediate interactions with RNAs in mRNA transport, storage, stability, and regulation of translation (Fig. 1A) (3, 10). Each KH domain has been reported to have a mutation either causing FXS or found in patients with intellectual disability (4, 5, 11). In neurons, FMRP associates with a subset of mRNAs and represses their translation both in the cell body and near synapses (12). Loss of repression of these mRNAs is associated with alterations in synaptic plasticity and dendritic spine dynamics thought to underlie...

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“…10 FMRP has been shown to use its RGG box domain to bind mRNA targets that form G-quadruplex structures. [11][12][13] A Gquadruplex is assembled from stacked G quartets that are formed by guanine residues arranged in a planar configuration by Hoogsteen-type hydrogen bonds.…”

Section: Introductionmentioning

confidence: 99%

FMRP interacts with G-quadruplex structures in the 3’-UTR of its dendritic target Shank1 mRNA

et al. 2014

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Fragile X syndrome (FXS), the most common cause of inherited intellectual disability, is caused by the loss of expression of the fragile X mental retardation protein (FMRP). FMRP, which regulates the transport and translation of specific mRNAs, uses its RGG box domain to bind mRNA targets that form G-quadruplex structures. One of the FMRP in vivo targets, Shank1 mRNA, encodes the master scaffold proteins of the postsynaptic density (PSD) which regulate the size and shape of dendritic spines because of their capacity to interact with many different PSD components. Due to their effect on spine morphology, altered translational regulation of Shank1 transcripts may contribute to the FXS pathology. We hypothesized that the FMRP interactions with Shank1 mRNA are mediated by the recognition of the G quadruplex structure, which has not been previously demonstrated. In this study we used biophysical techniques to analyze the Shank1 mRNA 3'-UTR and its interactions with FMRP and its phosphorylated mimic FMRP S500D. We found that the Shank1 mRNA 3 0 -UTR adopts two very stable intramolecular G-quadruplexes which are bound specifically and with high affinity by FMRP both in vitro and in vivo. These results suggest a role of G-quadruplex RNA motif as a structural element in the common mechanism of FMRP regulation of its dendritic mRNA targets.

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Interactions of the G quartet forming semaphorin 3F RNA with the RGG box domain of the fragile X protein family

Cited by 66 publications

References 49 publications

Vasa promotes Drosophila germline stem cell differentiation by activating mei-P26 translation by directly interacting with a (U)-rich motif in its 3′ UTR

Vasa promotes Drosophila germline stem cell differentiation by activating mei-P26 translation by directly interacting with a (U)-rich motif in its 3′ UTR

Crystal structure reveals specific recognition of a G-quadruplex RNA by a β-turn in the RGG motif of FMRP

FMRP interacts with G-quadruplex structures in the 3’-UTR of its dendritic target Shank1 mRNA

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