RNA LOCALIZATION

Global mRNA polarization regulatestranslation efficiency in theintestinal epitheliumAndreas E. Moor,1 Matan Golan,1 Efi E. Massasa,1 Doron Lemze,1 Tomer Weizman,1

Rom Shenhav,1 Shaked Baydatch,1 Orel Mizrahi,2 Roni Winkler,2 Ofra Golani,3

Noam Stern-Ginossar,2 Shalev Itzkovitz1*

Asymmetric messenger RNA (mRNA) localization facilitates efficient translation in cellssuch as neurons and fibroblasts. However, the extent and importance of mRNA polarizationin epithelial tissues are unclear. Here, we used single-molecule transcript imaging andsubcellular transcriptomics to uncover global apical-basal intracellular polarization of mRNA inthe mouse intestinal epithelium.The localization of mRNAs did not generally overlap proteinlocalization. Instead, ribosomes weremore abundant on the apical sides, and apical transcriptswere consequently more efficiently translated. Refeeding of fasted mice elicited a basal-to-apical shift in polarization of mRNAs encoding ribosomal proteins, which was associated witha specific boost in their translation.This led to increased protein production, required forefficient nutrient absorption.These findings reveal a posttranscriptional regulatory mechanisminvolving dynamic polarization of mRNA and polarized translation.

Intracellular mRNA localization is an impor-tant determinant of various cellular functions(1–6). In mammals, localized translation oftranscripts at the sites where their encodedproteins are needed is thought to confer cel-

lular efficiency and a timely response (2, 7, 8).Epithelial tissues are inherently polarized withdistinct functional specialization of basal andapical sides. The intestinal epithelium consists

of amonolayer of enterocytes that absorbnutrientsfrom the apical lumen and excrete them intothe blood stream from the basal sides. Key trans-porters are specifically localized in these twocompartments (9). Owing to the transient na-ture of nutrients in the gut, efficient and timelytranslation of such proteins might be impor-tant. The mRNA intracellular localization of afew enterocyte genes have been reported (10–12);

however, we lack a global characterization ofintracellular mRNA localization and its biologicalfunctions in epithelia.To obtain a transcriptome-wide viewofmRNA

localization in the intestinal epithelium, we iso-lated apical and basal subcellular fractions bylaser capture microdissection (Fig. 1A) and per-formed RNA sequencing (RNA-seq) (13). Almost30% of the most highly expressed 2000 tran-scripts, for which experimental noise levels werelower, were significantly polarized (Fig. 1B andtable S1). To validate the sequencing results, weperformed single-molecule fluorescence in situhybridization (smFISH) (14) on intact intestinaltissues for 14 genes that span different polariza-tion patterns (Fig. 1, B toD).We found an excellentcorrespondence between the apical bias of mRNAmeasured with smFISH and with the whole-transcriptome method (Spearman’s r = 0.97, P <2.2 × 10–16, Fig. 1D and table S2).Given the functional polarization of enterocytes,

we hypothesized that the observed global mRNAapical-basal polarization would match the loca-tion of the corresponding proteins. The most ap-ically enriched gene sets included several nutrienttransporter annotations, many of which encodeapically localized proteins (fig. S1). Notably, how-ever, the mRNAs that encode basolaterally local-ized proteins were strongly biased in localizationtoward the apical side, opposite to where their

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Fig. 1. Global analysis of mRNA polarization in the intestinal epithelium.(A) Laser capture microdissection of paired apical and basal fragments.(B) RNA-seq data of isolated subcellular areas. Insets show smFISH validationresults of transcripts of interest. Four outlier data points are omitted fromthe plot.Of the transcripts, 645of 9905 are significantly apical and 779 of 9905are significantly basal. Dashed vertical line separates the 2000 most highly

expressed transcripts, of which 392 genes are significantly apical and 194 aresignificantly basal. (C) smFISH staining of the apical Apob (red) and basalCyb5r3 (green) mRNA. (D) Strong correlation of smFISH quantifications andRNAseq data for 14 representative genes (Spearman’s r=0.97,P<2.2 × 10–16);dots and error bars represent median and 95% confidence interval ofsmFISH and mean and SE of RNAseq. All scale bars are 10 mm.

1Department of Molecular Cell Biology, Weizmann Institute ofScience, Rehovot, Israel. 2Department of Molecular Genetics,Weizmann Institute of Science, Rehovot, Israel. 3LifeSciences Core Facilities, Weizmann Institute of Science,Rehovot, Israel.*Corresponding author. Email: shalev.itzkovitz@weizmann.ac.il

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encoded proteins reside (Fig. 2, A and B, andfig. S2A). Examples of basolateral proteins withdiscordantly polarized apical mRNA included E-cadherin (Cdh1 gene, Fig. 2B) and integrins (fig.S2A). Thus, the global mRNA polarization in theintestinal epithelium does not generally correlatewith the localization of the encoded proteins.To obtain a broader view of the relative local-

ization of mRNAs and their encoded proteins, weperformed mass spectrometry of laser capturemicrodissected apical and basal subcellular frac-tions (Fig. 2C and table S3). The apical bias of pro-teins and of mRNA was weakly anticorrelated

(Spearman’s r = –0.12, P = 8 × 10–4). This dataset revealed additional examples of discordantlocalizations of proteins and their encodingmRNA,suchas thebasal protein sodium-potassiumATPase(adenosine triphosphatase), encoded by the highlyapical mRNA Atp1b1 (fig. S2A). Notably, the massspectrometry data revealed a significantly higherabundance of ribosomal proteins at the apical side(Fig. 2C and table S4) and mitochondrial proteinsat the basal sides (fig. S3 and table S4). ThemRNAsthat encode ribosomal proteinswere basally biased,again constituting a discordant set of genes, withopposite localizations of mRNA and their en-

coded proteins (Fig. 2C). Using smFISH, we iden-tified a highly significant apical enrichment ofribosomalRNA 18S and 28S (Fig. 2D and fig. S2B,both twofold, P < 2.2 × 10–16), thus validating theapically polarized localization of the translationalmachinery. To assess whether the increased con-centration of ribosomes on the apical sides yieldshigher translation at this subcellular compart-ment,we treatedmicewithO-propargyl-puromycin(OPP), a reporter that is efficiently incorporatedinto nascent peptides after a pulse-chase intra-peritoneal injection (15) (fig. S4). Nascent peptideswere significantly more abundant on the apical

Moor et al., Science 357, 1299–1303 (2017) 22 September 2017 2 of 5

Fig. 2. Translational machinery isasymmetrically distributed.(A) Ranking of gene sets that aresignificantly enriched on the apicalcell sides. (B) Costaining of thediscordantly localized basolateralprotein E-cadherin, encoded byCdh1 (gray staining) and its apicallylocalized mRNA (green dots).(C) Mass spectrometry results ofmicrodissected areas demonstrateapical enrichment of ribosomal pro-teins and a lack of positive correla-tion (Spearman’s r = –0.12, P =8 × 10–4). Blue, all genes; red, geneswith GO-Ribosome annotation.(D) Representative smFISH stainingand quantification of intracellulardistribution of rRNA 18S and 28S(n = 142 single cells,P < 2.2 × 10–16).(E) Representative staining andquantification of nascent proteins(n = 143 single cells,P < 2.2 × 10–16).(F) Normalized coverage plot ofribosome footprint sequencingdata (n = 3 mice, gray area denotesSD). (G) TE increases with theapical bias of genes in fastingmousesamples.The y axis shows themean TE over a sliding windowof 1000 genes consecutively shiftedfrom the most basal gene to themost apical gene, with a 500-geneoverlap.The x axis is themean apicalbias of genes within each window.Patches are standard errors of themean. (H) Translational efficiency ofthe significantly localized transcripts(false discovery rate adjustedP value <0.1, n apical = 346,n basal = 141, P = 8.4 × 10–10;20 outlier data points are omittedfrom the plot). Data include onlygenes for which we obtainedTE values and that are of epithelialorigin (methods). All scale barsare 10 mm.

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sides of the intestinal enterocytes (Fig. 2E, 1.82-fold higher concentration at the apical side, P <2.2 × 10–16). Thus, translation in the intestinalenterocytes is also strongly polarized, with a two-fold higher apical concentration of the transla-tional machinery.We next asked whether apical-basal mRNA

polarization could regulate translation efficiency(TE) in this tissue.Weperformedribosomeprofilingand RNA-seq of intestinal isolates from fastingmice and computed the TE of intestinal tran-scripts (Fig. 2F, fig. S5, and tables S5 and S6).Genes with significantly apical mRNA localiza-tion had almost twofold higher TE comparedto genes with significantly basal mRNA (Fig. 2,G and H, P = 8.36 × 10–10). In several nonpolar-izedmouse primary cell data sets, the apical andbasal groups were either indistinguishable or dis-

played much weaker apical enrichment of TE(fig. S6A). Thus, mRNA apico-basal polarizationmodulates TE in the intestinal epithelium.The intestinal lumen is a highly dynamicmicro-

environment that exhibits temporal oscillationin nutrient availability (16). Dynamic translocationofmRNAbetween the basal and apical sides couldpotentiallymodulate translational efficiency. Toexplore such dynamic responses, we performedour transcriptome-widemRNApolarizationmea-surements and ribosome profiling on duodenumtissues of fasting and refed mice (Fig. 3A). Thetranslational machinery remained apical after re-feeding, as evident by the apical polarization ofribosomal RNA (rRNA) (fig. S6B), and the TE ofgeneswas higher themore apically polarizedweretheir transcripts (fig. S6C). mRNAs that encoderibosomal proteins (RNA-RPs) shifted from the

basal localization in fasted intestines to a moreapical localization after 2 hours of refeeding (Fig.3A, 1.36-fold for RNA-RPs versus 1.08-fold forother transcripts, P = 0.0001). We also observed aconcomitant increase in TE for these transcriptsupon shifting their localization to the apical side—the side inwhich the translationmachinery ismoreabundant (Fig. 3A and fig. S7, 1.87-fold, P < 2 ×10–16).We validated these localization changes forRpl3 andRpl4mRNAby smFISH (Fig. 3, B andC,Rpl3 P = 1.8 × 10–8, Rpl4 P = 2.9 × 10–13). Thecellular mRNA expression levels of these geneswere not increased (fig. S8, A to C). After 4 hoursof refeeding, increased production of ribosomalcomponents was associated with significantlyhigher protein synthesis compared to the fastingstate (Fig. 3D, 2.05-fold, P < 2.2 × 10–16). TE ofbrush border proteins (17) also showed a slight,

Moor et al., Science 357, 1299–1303 (2017) 22 September 2017 3 of 5

Fig. 3. Dynamicshifts of localizedtranscripts areassociated withdifferential transla-tional efficiency.(A) Scatterplot of TEand mRNA localiza-tion changes whencomparing threefasting and threerefed mice (n = 6282transcripts).Thex axis is log2 of theratios between TEs inrefed and in fastingstates; the y axisis log2 of the ratiosof apical biasesbetween refed andfasting states, whereapical bias for eachcondition is the ratioof apical and basalTPM (methods).Upon refeeding,mRNAs encodingribosomal proteins(red dots) becomemore apically polar-ized and are trans-lated more efficiently.(B) smFISH imagesof Rpl3 and Rpl4mRNA across meta-bolic states.(C) Quantificationof Rpl3 and Rpl4smFISH analysesacross metabolicstates (n = 70 singlecells, Rpl3 P =1.8 × 10–8, Rpl4P = 2.9 × 10–13).(D) Comparisonof nascent proteincontent at fasting and5-hour refeeding time point (n = 188 single cells, P < 2.2 × 10–16). All scale bars are 10 mm.

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yet significant increase upon refeeding (1.1-fold,P = 0.02, fig. S8D). Thus, dynamic translocation ofmRNAs encoding ribosomal proteins to the moretranslationally active apical side is associatedwitha specific translation of ribosomal components,facilitating a burst of protein production to meetthe absorption demands upon refeeding.What facilitates this broadmRNApolarization?

Active transport by motor proteins along the

cytoskeleton, molecular anchors that bind andkeep transcripts localized, or spatially varyingRNA degradation rates within the cell have allbeen implicated in determining localization (1, 18).Additionally, our finding that the ribosomes areapically polarized in enterocytes could indicatethat preferential retention of highly translatedmRNA could lead to their apical polarization(1, 18). To investigate whether microtubules me-

diate the observed mRNA localization patterns,we depolymerized the microtubule network byinjecting mice intraperitoneally with nocodazole(Fig. 4, A and B). The basally polarized Net1,Cyb5r3, Rpl3, and Rpl4 transcripts lost polariza-tion, and the apically polarized Apob and Cdh17transcripts were significantly less apical (Fig. 4,A and B, and fig. S9). Although ribosomal RNAremained significantly apically polarized in

Moor et al., Science 357, 1299–1303 (2017) 22 September 2017 4 of 5

Fig. 4. IntestinalmRNA localizationis mediated by themicrotubule network.(A) Nocodazolestrongly perturbsthe polarization ofNet1 and Apob (Net1n = 94 cells, P = 1.1 ×10–15, Apob n = 96cells, P = 2.9 × 10–12).(B) RepresentativesmFISH imagesof strongly polarNet1 and Apob tran-scripts in vehicle- ornocodazole-injectedanimals. Scale bar,10 mm. (C) IntracellularmRNA localizationof Net1 and Apob inintestinal organoidsphenocopies in vivoobservations. High-lighted inset is shownin higher magnificationin top row of (D).Scale bar, 50 mm.(D) smFISH images ofNet1 and Apob innocodazole-,ispinesib-, or vehicle-treated organoids.Scale bar, 10 mm.(E) Single-cell quantifi-cation of the nocoda-zole and ispinesibeffects on transcriptlocalization in smFISHimages of intestinalorganoids (vehiclen = 101, ispinesibn = 68, nocodazole n =77 cells, costainingof Apob and Net1).(F) Transcripts thatencode ribosomalproteins are stored inthe less translationallyactive basal side of theintestinal epithelium infasting mice. Refeed-ing induces a trans-location of thesetranscripts into themore translationally active apical cell side.This translocation is associated with a concomitant increase in their translational efficiency.The increased ribosomalbiogenesis is reflected in an increased total protein synthesis.

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nocodazole-treated mice, the extent of rRNA po-larizationwas smaller compared to that in vehicle-treated controls (fig. S10). To assess whether thedifference in polarization of apical mRNA uponnocodazole treatment could stemfromcoordinatedmovement of ribosomes and their retainedmRNAto the basal side, we measured the combined po-larization of both rRNA and of four apical geneswith high TE in nocodazole-treated mice and incontrols, and stratified the results according tothe single-cell rRNA polarization. Microtubuledisruption decreased the apical polarization ofthese genes even when controlling for rRNApolarization changes (figs. S10 and S11A). Thus,themicrotubulenetworkmediates the asymmetriclocalization of these and potentially other tran-scripts in the intestinal epithelium, whereas ad-ditional anchoring mechanisms seem to renderthe apical localization of ribosomes less sensi-tive to microtubule disruption. Selective inhibi-tion of kinesin 5 in intestinal organoids (19, 20)using ispinesib (21) decreased the polarizationof Apob and Net1 (Fig. 4, C to E) but not ofCdh17 and Pigr (fig. S9, E and F).Given the transient nature of inputs in the gut,

consistently high translation rates could be ener-getically inefficient (22). When nutrients arrive,however, there is a short temporal window inwhich they must be efficiently absorbed (9).Nutrient availability was shown to stimulaterapid ribosome biogenesis in other contexts(23, 24) via activation of translation initiationfactors, rRNA transcription, and translation ofRNAs that encode the translational machinery(24). Here, we found a similar burst of proteintranslation upon refeeding, associated with arapid translocation of ribosomal-protein encod-ing transcripts, the nature of which is yet to bedetermined, into the translationally active apical

side (Fig. 4F). The burst of ribosomal proteintranslation upon refeeding resembles a “bang-bang” control strategy, in which resources arefirst invested inmaking the ribosomal “machines,”to facilitate a rapid increase in total protein output(25, 26). Our approach, combining laser-capturemicrodissectionandwhole-transcriptomesequenc-ing with smFISH, can be readily applied to char-acterize mRNA polarization in other tissuesand organisms. Future studies will determine ifmRNA mislocalizations are causatively involvedin pathophysiology.

REFERENCES AND NOTES

1. A. R. Buxbaum, G. Haimovich, R. H. Singer, Nat. Rev. Mol. CellBiol. 16, 95–109 (2015).

2. C. E. Holt, S. L. Bullock, Science 326, 1212–1216 (2009).3. K. C. Martin, A. Ephrussi, Cell 136, 719–730 (2009).4. E. Lécuyer et al., Cell 131, 174–187 (2007).5. S. Mili, K. Moissoglu, I. G. Macara, Nature 453, 115–119 (2008).6. H. Jambor et al., eLife 4, e05003 (2015).7. F. Besse, A. Ephrussi, Nat. Rev. Mol. Cell Biol. 9, 971–980

(2008).8. A. J. Rodriguez, K. Czaplinski, J. S. Condeelis, R. H. Singer,

Curr. Opin. Cell Biol. 20, 144–149 (2008).9. K. N. Frayn, Metabolic Regulation: A Human Perspective

(Wiley-Blackwell, Chichester, UK, ed. 3, 2010).10. E. H. H. M. Rings et al., FEBS Lett. 300, 183–187 (1992).11. J. A. Barth et al., J. Histochem. Cytochem. 46, 335–343 (1998).12. W. Li, S. D. Krasinski, M. Verhave, R. K. Montgomery,

R. J. Grand, Gastroenterology 115, 86–92 (1998).13. S. Nichterwitz et al., Nat. Commun. 7, 12139 (2016).14. A. Raj, P. van den Bogaard, S. A. Rifkin, A. van Oudenaarden,

S. Tyagi, Nat. Methods 5, 877–879 (2008).15. J. Liu, Y. Xu, D. Stoleru, A. Salic, Proc. Natl. Acad. Sci. U.S.A.

109, 413–418 (2012).16. C. A. Thaiss et al., Cell 167, 1495–1510.e12 (2016).17. R. E. McConnell, A. E. Benesh, S. Mao, D. L. Tabb, M. J. Tyska,

Am. J. Physiol. Gastrointest. Liver Physiol. 300, G914–G926(2011).

18. C. A. Pratt, K. L. Mowry, Curr. Opin. Cell Biol. 25, 99–106 (2013).19. T. Sato et al., Nature 459, 262–265 (2009).20. T. Sato et al., Gastroenterology 141, 1762–1772 (2011).21. L. Lad et al., Biochemistry 47, 3576–3585 (2008).22. D. F. Rolfe, G. C. Brown, Physiol. Rev. 77, 731–758 (1997).

23. C. Mayer, I. Grummt, Oncogene 25, 6384–6391 (2006).24. M. Laplante, D. M. Sabatini, Cell 149, 274–293 (2012).25. S. Itzkovitz, I. C. Blat, T. Jacks, H. Clevers, A. van Oudenaarden,

Cell 148, 608–619 (2012).26. D. Madar et al., BMC Syst. Biol. 7, 136 (2013).

ACKNOWLEDGMENTS

We thank K. Bahar Halpern, B. Toth, and S. Ben-Moshe forvaluable comments on the manuscript. We thank the L Lokeyanimal facility (Weizmann Institute of Science, Rehovot, Israel), theCrown Institute for Genomics (Weizmann Institute of Science,Rehovot, Israel), and the Smoler Protein Research Center(Technion, Haifa, Israel) for help with experimental procedures.A.E.M. is supported by the Swiss National Science Foundation(grant 158999) and the European Molecular Biology Organization(EMBO) Long-Term Fellowship program (ALTF 306-2016).N.S.-G. research is funded by the European Research Councilstarting grant (StG-2014-638142). N.S.-G. is incumbent of theSkirball Career Development Chair in New Scientists. S.I. issupported by the Henry Chanoch Krenter Institute for BiomedicalImaging and Genomics, The Leir Charitable Foundations,Richard Jakubskind Laboratory of Systems Biology,Cymerman-Jakubskind Prize, The Lord Sieff of Brimpton MemorialFund, the I-CORE program of the Planning and BudgetingCommittee and the Israel Science Foundation (grants 1902/12and 1796/12), the Israel Science Foundation grant no. 1486/16,the EMBO Young Investigator Program, and the EuropeanResearch Council under the European Union’s Seventh FrameworkProgramme (FP7/2007-2013)–ERC grant agreement no. 335122.S.I. is the incumbent of the Philip Harris and Gerald RonsonCareer Development Chair. All data supporting the findingsof this study and all analysis codes are available within thearticle and its supplementary materials or from the correspondingauthor upon request. The generated sequencing data havebeen deposited in the GenBank Gene Expression Omnibus database(www.ncbi.nlm.nih.gov/geo/) under accession code GSE95416.

SUPPLEMENTARY MATERIALS

www.sciencemag.org/content/357/6357/1299/suppl/DC1Materials and MethodsFigs. S1 to S11Tables S1 to S7References (27–52)

16 March 2017; resubmitted 3 July 2017Accepted 1 August 2017Published online 10 August 201710.1126/science.aan2399

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Global mRNA polarization regulates translation efficiency in the intestinal epithelium

Mizrahi, Roni Winkler, Ofra Golani, Noam Stern-Ginossar and Shalev ItzkovitzAndreas E. Moor, Matan Golan, Efi E. Massasa, Doron Lemze, Tomer Weizman, Rom Shenhav, Shaked Baydatch, Orel

originally published online August 10, 2017DOI: 10.1126/science.aan2399 (6357), 1299-1303.357Science 

, this issue p. 1299; see also p. 1235Sciencetranslational efficiency in the intestinal epithelium.shift in localization and a boost in translation. Thus, dynamic polarization of mRNA and polarized translation modulate efficient translation. On refeeding of fasted mice, gut cell mRNAs encoding ribosomal proteins exhibit a basal-to-apicallocalization does not generally overlap protein localization; instead, ribosomes are apically biased, which allows more

mRNAenterocytes tend to distribute to the cells' apical or basal cell sides (see the Perspective by Gáspár and Ephrussi). found that transcripts in intestinalet al.known in several cell types but is poorly understood in gut epithelial cells. Moor

The distribution of RNA in cells is important for efficient translation into proteins. Asymmetric RNA localization isLocation, location, location

ARTICLE TOOLS http://science.sciencemag.org/content/357/6357/1299

MATERIALSSUPPLEMENTARY http://science.sciencemag.org/content/suppl/2017/08/09/science.aan2399.DC1

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