Recurrent Motifs Research Articles

Effects of Noncanonical Base Pairing on RNA Folding: Structural Context and Spatial Arrangements of G·A Pairs.

Noncanonical base pairs play important roles in assembling the three-dimensional structures critical to the diverse functions of RNA. These associations contribute to the looped segments that intersperse the canonical double-helical elements within folded, globular RNA molecules. They stitch together various structural elements, serve as recognition elements for other molecules, and act as sites of intrinsic stiffness or deformability. This work takes advantage of new software (DSSR) designed to streamline the analysis and annotation of RNA three-dimensional structures. The multiscale structural information gathered for individual molecules, combined with the growing number of unique, well-resolved RNA structures, makes it possible to examine the collective features deeply and to uncover previously unrecognized patterns of chain organization. Here we focus on a subset of noncanonical base pairs involving guanine and adenine and the links between their modes of association, secondary structural context, and contributions to tertiary folding. The rigorous descriptions of base-pair geometry that we employ facilitate characterization of recurrent geometric motifs and the structural settings in which these arrangements occur. Moreover, the numerical parameters hint at the natural motions of the interacting bases and the pathways likely to connect different spatial forms. We draw attention to higher-order multiplexes involving two or more G·A pairs and the roles these associations appear to play in bridging different secondary structural units. The collective data reveal pairing propensities in base organization, secondary structural context, and deformability and serve as a starting point for further multiscale investigations and/or simulations of RNA folding.

Atomic Motif Recognition in (Bio)Polymers: Benchmarks From the Protein Data Bank.

Rationalizing the structure and structure–property relations for complex materials such as polymers or biomolecules relies heavily on the identification of local atomic motifs, e.g., hydrogen bonds and secondary structure patterns, that are seen as building blocks of more complex supramolecular and mesoscopic structures. Over the past few decades, several automated procedures have been developed to identify these motifs in proteins given the atomic structure. Being based on a very precise understanding of the specific interactions, these heuristic criteria formulate the question in a way that implies the answer, by defining a list of motifs based on those that are known to be naturally occurring. This makes them less likely to identify unexpected phenomena, such as the occurrence of recurrent motifs in disordered segments of proteins, and less suitable to be applied to different polymers whose structure is not driven by hydrogen bonds, or even to polypeptides when appearing in unusual, non-biological conditions. Here we discuss how unsupervised machine learning schemes can be used to recognize patterns based exclusively on the frequency with which different motifs occur, taking high-resolution structures from the Protein Data Bank as benchmarks. We first discuss the application of a density-based motif recognition scheme in combination with traditional representations of protein structure (namely, interatomic distances and backbone dihedrals). Then, we proceed one step further toward an entirely unbiased scheme by using as input a structural representation based on the atomic density and by employing supervised classification to objectively assess the role played by the representation in determining the nature of atomic-scale patterns.

A Recurrent Motif: Diversity and Evolution of ShKT Domain Containing Proteins in the Vampire Snail Cumia reticulata.

Proteins of the ShK superfamily are characterized by a small conserved domain (ShKT), first discovered in small venom peptides produced by sea anemones, and acting as specific inhibitors of voltage-dependent and calcium-activated K+ channels. The ShK superfamily includes both small toxic peptides and larger multifunctional proteins with various functions. ShK toxins are often important components of animal venoms, where they perform different biological functions including neurotoxic and immunosuppressive effects. Given their high specificity and effectiveness, they are currently regarded as promising pharmacological lead compounds for the treatment of autoimmune diseases. Here, we report on the molecular analysis of ShKT domain containing proteins produced by the Mediterranean vampire snail Cumia reticulata, an ectoparasitic gastropod that feeds on benthic fishes. The high specificity of expression of most ShK transcripts in salivary glands identifies them as relevant components of C. reticulata venom. These ShK proteins display various structural architectures, being produced either as single-domain secretory peptides, or as larger proteins combining the ShKT with M12 or CAP domains. Both ShKT-containing genes and their internal ShKT domains undergo frequent duplication events in C. reticulata, ensuring a high level of variability that is likely to play a role in increasing the range of their potential molecular targets.

Co-crystals of an organic triselenocyanate with ditopic Lewis bases: recurrent chalcogen bond interactions motifs.

Organic selenocyanates R-Se-CN can act as an amphoteric chalcogen bond (ChB) donor (through the Se atom) and acceptor (through the N atom lone pair). Co-crystallization of tri-substituted 1,3,5-tris(selenocyanatomethyl)-2,4,6-trimethylbenzene (1) is investigated with different ditopic Lewis bases acting as chalcogen bond (ChB) acceptors to investigate the outcome of the competition, as ChB acceptor, between the nitrogen lone pair of the SeCN group and other Lewis bases involving pyridinyl or carbonyl functions. In the presence of tetramethylpyrazine (TMP), benzoquinone (BQ) and para-dinitrobenzene (pDNB) as ditopic Lewis bases, a recurrent oligomeric motif stabilized by six ChB interactions is observed, involving six SeCN groups and the ChB acceptor sites of TMP, BQ and pDNB in the 2:1 adducts (1)2·TMP, (1)2·BQ and (1)2·pDNB.

Cannibal Plants: Tropiques and Martinican Aesthetics

The "cultural journal" Tropiques was published on the island of Martinique during the regime of the collaborationist Vichy government. Contributors to this journal found themselves limited in the content they could publish by right-wing government censors whose official position encouraged the expression of a certain amount of "regionalism," as this was considered by the regime. One recurrent motif was the topic of plants—a seemingly benign topic, but one which, when read as a dialog between the contributors, both subverts tropes associated with flora and shows an emerging sense of what could be a Martinican or Antillean identity. This dialog consisted in texts ranging from botanical descriptions to probably the most well-known example: an article by Suzanne Césaire called "L'Homme plante," where she uses ethnographer Leo Frobenius's image of the "plant man" to describe the character of the Martinican people. This image seems to evoke passivity, but in the end, it is not only a complex metaphor for a model of autonomy and resistance for the individual Martinican, but also develops a sense of the emplacement of the population as a whole in the geography of the Antilles and it is a concrete emblem for the struggle for alimentary and economic autonomy exacerbated under the wartime blockade.

Late Poems: “One of the Inhabitants of the West” and “Nuns Painting Water-Lilies”

Late Poems: “One of the Inhabitants of the West” and “Nuns Painting Water-Lilies” Bart Eeckhout, Juliette Utard, Charles Altieri, Massimo Bacigalupo, Lisa Goldfarb, Gül Bilge Han, Glen MacLeod, Rachel Malkin, Maureen N. McLane, Edward Ragg, and Tony Sharpe PREFATORY NOTE: The transcript of this sixth and final seminar, which took place on June 21, 2017, in Bogliasco, Italy, uses the following abbreviations: BE (Bart Eeckhout, University of Antwerp & Netherlands Institute for Advanced Study), JU (Juliette Utard, Sorbonne University & CNRS), CA (Charles Altieri, University of California, Berkeley), MB (Massimo Bacigalupo, University of Genoa), LG (Lisa Goldfarb, Gallatin School, New York University), GH (Gül Bilge Han, Stockholm University), GM (Glen MacLeod, University of Connecticut), RM (Rachel Malkin, University of Oxford), MM (Maureen N. McLane, New York University), ER (Edward Ragg, Beijing), TS (Tony Sharpe, Lancaster University). The transcript was heavily edited for readability and circulated among participants for fine-tuning; it thus reproduces the dynamic of exchange without in any way striving to be literal. Readers who come upon this material outside the special issue to which it belongs are advised to read the editorial introduction for an account of the rationale behind the following discussion. A Negative Epiphany about Buried Guilt BE: For this final seminar, Juliette has accepted to kick off the discussions before I append a few comments of my own. But let me start by reading out the first poem, “One of the Inhabitants of the West”: Our divinations,Mechanisms of angelic thought,The means of prophecy, [End Page 112] Alert us mostAt evening’s one starAnd its pastoral text, When the establishmentsOf wind and light and cloudAwait an arrival, A reader of the text,A reader without a body,Who reads quietly: “Horrid figures of Medusa,These accents explicateThe sparkling fall of nightOn Europe, to the last Alp,And the sheeted Atlantic. These are not banlieusLacking men of stone,In a well-rosed two-lightOf their own.I am the archangel of evening and praise This one star’s blaze.Suppose it was a drop of blood . . .So much guilt lies buriedBeneath the innocenceOf autumn days.” (CPP 428–29) JU: This poem was first published in December 1952 in The Nation. Eleanor Cook calls it “Another angel poem, another reader poem, another poem meditating on day’s end, here as if on the edge of Western Europe. If it is touched by any sense of ‘Westward the course of empire’ . . . there is no sense of triumph” (280). I’m interested in the poem’s apparent sense of tedium, its rehearsal of various old themes, at a moment when Stevens was attempting to develop a distinct late style in The Rock. Tedium is a recurrent motif in that final section of The Collected Poems, from “An Old Man Asleep” to “Song of Fixed Accord” and “Long and Sluggish Lines.” The question to which Stevens seems to return is, “What difference can one ultimately make?” As he says in “Long and Sluggish Lines,” “at so much more / Than seventy, where one looks, one has been there before” (CPP 442). There is an unresolved tension in the poem between an effort toward singularity (“One of the,” “This one star’s blaze”) and an acknowledgment [End Page 113] of commonality (the poem, after all, begins in the first-person plural). So, like Cook, I treat the titular West not as troping conquest and the frontier but rather the setting sun and evening star—what Stevens in his previous volume called “The Westwardness of Everything” (CPP 389). Hence perhaps the figure of the Medusa as threatening petrification—which makes perfect sense in a collection entitled The Rock. To this cluster of images may be added the “fall of night / On Europe,” that strangely “last Alp” (as if the Alps were countable and we had reached the end of a series), the corpse-like “sheeted Atlantic,” shrouded in rigor mortis, the “drop of blood,” and then the final confession of “guilt.” “One of the Inhabitants of the West” seems to address finitude, but in a minor key. It is rather uncharacteristic of Stevens’s late...

Recovering from Amnesia in Octavia Butler’s Texts

EnThree novels by Octavia Butler, the first famous African-American speculative fiction writer, feature characters who suffer from memory loss. All three novels begin with the isolated and forgetful individual who in the course of the narrative recovers some memory of the past and uses it to build or reconstruct family ties. The article analyzes the way those amnesiac subjects are represented and the types of amnesia they suffer from. It also looks at some elements of what is retrieved in order to trace back the roots of certain recurrent motifs. Finally, it questions the figure of the Black woman fighting to recover her memory and use it as basis to construct a community.

Армянский эпос о неистовых сасунцах и индийская «Махабхарата»: сходство этнографического субстрата

The author summarizes the results of his search for parallels between the Armenian epic “Sasna cṙer” (“Daredevils of Sassoun”) and the Mahābhārata. The comparative study has revealed considerable similarity in the “ethnographic substratum” of both epics, particularly that relating to the archaic social organization mirrored by the epic. The earliest layer of both the Armenian and the Indian epics retains the memory of a rural, largely pastoral society, in which an important role was played by the fraternities of young warriors. In the Armenian epic, this is indicated by recurrent motifs such as the young heroes’ rampage followed by exile, the foundation of their own outpost in the backwoods, young male warriors’ fraternities, their defense of herds, warding off enemy attacks, battle frenzy (a common characteristic of all the Sasun heroes), their immutable tutor and leader (“uncle”) Keri Toros, allusions to orgiastic feasts, premarital freedom enjoyed by boys and girls, etc. In Armenians, these motifs were supported by the existence until the recent times of age sets, described by ethnographers. The comparative study of the Armenian epic reveals its hitherto unnoticed socio-historical aspects. Its wider use for studying other epic traditions (not only Indo-European but also those of other peoples inhabiting the Caucasus and the Eurasian steppes) will contribute to the comparative epic studies.

Paesaggi dell'anima: su alcuni interni della letteratura francese

Landscapes of the Soul: On Some Home Interiors in French LiteratureBy means of an analysis of architectural features and furnishing elementspresent in a number of passages from French literature (such asthe most innovative French epistolary novel, Rousseau’s La NouvelleHeloise, and two poems by Baudelaire and Mallarme), this essay aimsto highlight recurrent motifs that reveal unforeseen consonances andresonances within the described environments (two dining rooms andone bedroom). The same motifs form a picture which reveals an intensifyingprocess of spoliation of the concrete elements present in therooms, as well as a marked loss in functionality. Such spoliation andloss are shown to be directly proportional to symbolic intensity and poignance.If the presence of an emotive connection between those placesand originary feelings related to fragile, lost, or unrealizable happinessremains constant, the poetical treatment of such themes becomes increasinglyrigorous and enigmatic.The piece is indebted on the one hand to a profoundly inspiring bookby Guido Paduano, Il testo e il mondo, and on the other to a sharedexperience of feelings in some ways related to the ones analyzed in thearticle, feelings forever intertwined with the memory of the author’sfriend Tiziana Goruppi.

Ab Initio Predictions of Double-Layer TiO2-Terminated SrTiO3(001) Surface Reconstructions

Predicting oxide surface structures, and surface structures in general, is a significant science frontier. Herein, we demonstrate an Ising model approach combined with density-functional calculations to predict several SrTiO3(001) surface reconstructions. Constrained Ising tilings of TiO5□ truncated octahedral units are used to construct the TiO2 adlayers of SrTiO3(001) surface reconstructions with TiO2 double-layer terminations for a given periodicity and composition. This method is used to re-examine the (√5 × √5)R26.6° reconstruction and to predict several (√10 × √10)R18.4° reconstructions. The surface structures and energies of the newly predicted reconstructions, along with consideration of the previously reported (√13 × √13)R33.7°, 3 × 3, and (√2 × √2)R45.0° reconstructions, indicate that the larger, more complex TiO2 double-layer SrTiO3(001) reconstructions with lower coverages of the TiO2 adlayer are made up of recurrent structural motifs represented by smaller, simpler reconstructions with higher...

Asymmetric vasopressin signaling spatially organizes the master circadian clock.

The suprachiasmatic nucleus (SCN) is the neural network that drives daily rhythms in behavior and physiology. The SCN encodes environmental changes through the phasing of cellular rhythms across its anteroposterior axis, but it remains unknown what signaling mechanisms regulate clock function along this axis. Here we demonstrate that arginine vasopressin (AVP) signaling organizes the SCN into distinct anteroposterior domains. Spatial mapping of SCN gene expression using in situ hybridization delineated anterior and posterior domains for AVP signaling components, including complementary patterns of V1a and V1b expression that suggest different roles for these two AVP receptors. Similarly, anteroposterior patterning of transcripts involved in Vasoactive Intestinal Polypeptide- and Prokineticin2 signaling was evident across the SCN. Using bioluminescence imaging, we then revealed that inhibiting V1A and V1B signaling alters period and phase differentially along the anteroposterior SCN. V1 antagonism lengthened period the most in the anterior SCN, whereas changes in phase were largest in the posterior SCN. Further, separately antagonizing V1A and V1B signaling modulated SCN function in a manner that mapped onto anteroposterior expression patterns. Lastly, V1 antagonism influenced SCN period and phase along the dorsoventral axis, complementing effects on the anteroposterior axis. Together, these results indicate that AVP signaling modulates SCN period and phase in a spatially specific manner, which is expected to influence how the master clock interacts with downstream tissues and responds to environmental changes. More generally, we reveal anteroposterior asymmetry in neuropeptide signaling as a recurrent organizational motif that likely influences neural computations in the SCN clock network.

Local structural motifs in proteins: Detection and characterization of fragments inserted in helices

The global/local fold of protein structures is stabilized by a variety of specific interactions. A primary role in this context is played by hydrogen bonds. In order to identify novel motifs in proteins, we searched Protein Data Bank structures looking for backbone H-bonds formed by NH groups of two (or more) consecutive residues with consecutive CO groups of distant residues in the sequence. The present analysis unravels the occurrence of recurrent structural motifs that, to the best of our knowledge, had not been characterized in literature. Indeed, these H-bonding patterns are found (i) in a specific parallel β-sheet capping, (ii) in linking of β-hairpins to α-helices, and (iii) in α-helix insertions. Interestingly, structural analyses of these motifs indicate that Gly residues frequently occupy prominent positions. The formation of these motifs is likely favored by the limited propensity of Gly to be embodied in helices/sheets. Of particular interest is the motif corresponding to insertions in helices that was detected in 1% of analyzed structures. Inserted fragments may assume different structures and aminoacid compositions and usually display diversified evolutionary conservation. Since inserted regions are physically separated from the rest of the protein structure, they represent hot spots for ad-hoc protein functionalization.

Myoglobin‐Catalyzed C−H Functionalization of Unprotected Indoles

AbstractFunctionalized indoles are recurrent motifs in bioactive natural products and pharmaceuticals. While transition metal‐catalyzed carbene transfer has provided an attractive route to afford C3‐functionalized indoles, these protocols are viable only in the presence of N‐protected indoles, owing to competition from the more facile N−H insertion reaction. Herein, a biocatalytic strategy for enabling the direct C−H functionalization of unprotected indoles is reported. Engineered variants of myoglobin provide efficient biocatalysts for this reaction, which has no precedents in the biological world, enabling the transformation of a broad range of indoles in the presence of ethyl α‐diazoacetate to give the corresponding C3‐functionalized derivatives in high conversion yields and excellent chemoselectivity. This strategy could be exploited to develop a concise chemoenzymatic route to afford the nonsteroidal anti‐inflammatory drug indomethacin.

Things “Never Heard of Before”: Hogg, the Unnatural, and Romantic Cultural Science

ABSTRACTThis paper reads a recurrent motif of Hogg’s fiction—the unnatural—in relation to the “organic” cultural science propounded by some of Hogg’s contemporaries. For many Romantic commentators, social organizations like the rustic community, the ethnic group, and the nation bore the imprint of a symbiotic relationship between laws in ethnography and those of organic life. Hogg’s writing evokes scenarios in which such laws lose durability and fail to generate connections between land and community, self and other. In these moments, Hogg evokes the incongruous and fragmentary qualities of a native cultural experience, an otherness often erased from more dominant ethnographic and aesthetic depictions of Scotland’s cultural life.

Mela: festival scenes in South Asian cinema

ABSTRACTThis article examines a recurrent film motif across a number of South Asian films, mostly called Mela. It also offers some observations on melas, actual and allegorical, as represented in films but often seeming to exceed their containment in context so as to say more about the conviviality of life, where this is at issue, where life is at a juncture in need of resolution within the cycle of becoming. The issues of violence, loss, national identity, politics of interpretation and repetition in ideology are canvassed. While the essay is focused upon Mela films themselves, and South Asian film more broadly, it has of course been important to note work by scholars such as M. Madhava Prasad, Ashish Rajadhyaksha, Anjali Gera Roy, Tejaswini Niranjana, Gayatri Chakravorty Spivak and the help of my students, some of whom are named below.

Shaping Modern Shanghai: Colonialism in China's Global City by Isabella Jackson

Reviewed by: Shaping Modern Shanghai: Colonialism in China's Global City by Isabella Jackson Cécile Armand Isabella Jackson. Shaping Modern Shanghai: Colonialism in China's Global City. Cambridge: Cambridge University Press, 2018. 274 pp. $99.99 (cloth). Shaping Modern Shanghai: Colonialism in China's Global City offers a detailed study of the major colonial institution in one of the most important cities in Republican China: the Shanghai Municipal Council (SMC), which governed the International Settlement in Shanghai from the mid-1840s until the relinquishment of the settlement in May 1943. As a treaty port open to foreign residents and companies, Shanghai became not only a major industrial and financial hub in China but also a center for political and cultural life, playing a significant role in the birth of Chinese nationalism. At a time of economic development and political and social unrest (the May Thirtieth movement of 1925, recurrent strikes and boycotts, the Second Sino-Japanese War from 1937 to 1945), the SMC's primary mission was to maintain a stable environment that was conducive to business. As the population of the International Settlement grew from 100,000 in 1880 to 1,000,000 in 1930, the SMC expanded its areas of intervention and increasingly shaped everyday life in the International Settlement. Jackson's main argument is that the SMC exemplified the specifically transnational nature of colonialism in China. It was transnational in the sense that it involved nonstate actors who cooperated on the ground beyond national interests. The SMC was a transnational institution both politically and socially. Politically, it remained distant from imperial oversight. In contrast to the French Municipal Council that ruled the French Concession, the SMC was not subject to consular authority. The SMC, financially self-sufficient and able to maintain its own defense force, was proud of its political autonomy. In sum, the SMC acted much like the government of an independent city-state through its policy of territorial expansion and its governmental powers of taxing and policing the International Settlement. Socially, the SMC was a transnational body of people, including 25 different nationalities. Transnationalism, however, did not mean equality, as the SMC maintained a strict racial hierarchy between foreigners and Chinese, and even among foreigners (Russians, Sikhs). Transnational colonialism, Jackson argues, does not bear the positive assumption of cosmopolitanism. Although Chinese representation on the SMC increased after 1927, the British continued to dominate. Observing the peculiar neglect of the SMC in the history of colonialism in China, Jackson helps to fill this historiographical void by drawing on "the vast holdings of the Shanghai municipal archives" (and secondarily, those of the Foreign Office in Britain) and putting forward "the concept of transnational colonialism to explain the nature of colonialism in China" (2). This concept, Jackson argues, is not only more appropriate to the Shanghai case but is further applicable to other colonial contexts. Contributing to the [End Page E-25] recent scholarship on transnationalism, her argument has broad implications for the history of colonialism and nationalism in China and beyond. She offers a convincing alternative to such controversial concepts as hypo-, hyper-, semi-, and quasi-colonialism and the even more vague notion of "colonial modernity," all of which fail to capture the full complexity of colonialism in China. In contrast, she provides a more nuanced and embodied view of colonialism in Shanghai. As a social historian, her work is strongly grounded in primary materials and historical facts. She demonstrates a genuine concern for human actors, unveiling their daily work and the SMC's concrete impact on the residents' everyday lives. The book interweaves three key themes that serve as recurrent motifs throughout the book: similarities to and differences from British formal colonies, transnational influences and limits of transnationalism due to racial tensions, the challenges facing the SMC in the form of financial (tight budget), geographical (settlement boundaries), and legal (land regulations, bylaws) limitations. The book consists of five chapters—each one dealing with a particular area of municipal intervention—arranged according to their relative importance: finances, councilors and staff, police and conflict, public health and hygiene, and industrial, welfare, and social reforms. While the argument does not follow a chronological order, Jackson does...

Chalcogen bonding interactions in organic selenocyanates: from cooperativity to chelation

Organic selenocyanates form recurrent chain-like motifs ⋯Se(R)–CN⋯Se(R)–CN⋯ through short and linear chalcogen bonding Se⋯NC interactions. A chelating motif is also observed in a DMF solvate with two neighboring CH2–SeCN groups linked to the DMF oxygen atom.

Recurrent supramolecular motifs in discrete complexes and coordination polymers based on mercury halides: prevalence of chelate ring stacking and substituent effects

We report the synthesis, X-ray characterization and DFT study of five Hg(ii) complexes with Schiff bases containing a nicotinohydrazide core to explore the formation of chelate-ring π-stacking interactions.

Ironbark and stone

This article discusses Sunshine Coast writer Inga Simpson’s nature writing in three recent novels, Mr Wigg (2013), Nest (2014b) and Where the Trees Were (2016c). It addresses Simpson’s self-categorisation as a nature writer, and shows how the recurrent motif of sacred trees allows three introspective protagonists to reach new understandings of universal themes: loss of love and innocence, ageing, inheritance, childlessness, sexuality, death, ancient cultures, cultural integrity and preservation of the environment. The article considers Simpson’s ‘anti-Gothic’ approach to landscape in her novels, yet also shows how her ‘realist’ depictions of place evoke unease surrounding the issue of white belonging in Australia. Simpson’s metaphoric self-identification with trees, particularly theAustralian ironbark, is pivotal to the quiet power of her fiction’s exploration of belonging in the Australian landscape.

Phosphoproteomics reveals that glycogen synthase kinase-3 phosphorylates multiple splicing factors and is associated with alternative splicing

Glycogen synthase kinase-3 (GSK-3) is a constitutively active, ubiquitously expressed protein kinase that regulates multiple signaling pathways. In vitro kinase assays and genetic and pharmacological manipulations of GSK-3 have identified more than 100 putative GSK-3 substrates in diverse cell types. Many more have been predicted on the basis of a recurrent GSK-3 consensus motif ((pS/pT)XXX(S/T)), but this prediction has not been tested by analyzing the GSK-3 phosphoproteome. Using stable isotope labeling of amino acids in culture (SILAC) and MS techniques to analyze the repertoire of GSK-3-dependent phosphorylation in mouse embryonic stem cells (ESCs), we found that ∼2.4% of (pS/pT)XXX(S/T) sites are phosphorylated in a GSK-3-dependent manner. A comparison of WT and Gsk3a;Gsk3b knock-out (Gsk3 DKO) ESCs revealed prominent GSK-3-dependent phosphorylation of multiple splicing factors and regulators of RNA biosynthesis as well as proteins that regulate transcription, translation, and cell division. Gsk3 DKO reduced phosphorylation of the splicing factors RBM8A, SRSF9, and PSF as well as the nucleolar proteins NPM1 and PHF6, and recombinant GSK-3β phosphorylated these proteins in vitro RNA-Seq of WT and Gsk3 DKO ESCs identified ∼190 genes that are alternatively spliced in a GSK-3-dependent manner, supporting a broad role for GSK-3 in regulating alternative splicing. The MS data also identified posttranscriptional regulation of protein abundance by GSK-3, with ∼47 proteins (1.4%) whose levels increased and ∼78 (2.4%) whose levels decreased in the absence of GSK-3. This study provides the first unbiased analysis of the GSK-3 phosphoproteome and strong evidence that GSK-3 broadly regulates alternative splicing.