Region-Specific Activation of mRNA Translation by Inhibition of Bruno-Mediated Repression
Proteins are often enriched to specific regions within cells via localization of mRNAs. This phenomenon serves a variety of roles, both bringing together factors involved in particular cellular processes to enhance their efficiency, and in restricting proteins that could do harm if deployed at inappropriate positions. In the latter situation, translational repression prevents expression before mRNA localization, and there must be activation mechanisms to inhibit or override repression. How the processes of mRNA localization and translation are coordinated is not well understood, in part because cellular extracts prepared to study mechanisms in vitro do not retain the spatial information present in the intact cell. We developed an in vivo assay to monitor the pattern of translation in the Drosophila oocyte, where several patterning determinants must be localized to specific regions. Using this assay, we showed that repression of translation by the Bruno protein is inhibited, and we could visualize when and where this occurs during oogenesis. Regional activation occurs not only at the site of mRNA localization, but more broadly in a graded fashion, and it does not require an activation element in the mRNA. We also show that Bruno dimerizes, and that dimerization is important for translational activation.
Vyšlo v časopise:
Region-Specific Activation of mRNA Translation by Inhibition of Bruno-Mediated Repression. PLoS Genet 11(2): e32767. doi:10.1371/journal.pgen.1004992
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1004992
Souhrn
Proteins are often enriched to specific regions within cells via localization of mRNAs. This phenomenon serves a variety of roles, both bringing together factors involved in particular cellular processes to enhance their efficiency, and in restricting proteins that could do harm if deployed at inappropriate positions. In the latter situation, translational repression prevents expression before mRNA localization, and there must be activation mechanisms to inhibit or override repression. How the processes of mRNA localization and translation are coordinated is not well understood, in part because cellular extracts prepared to study mechanisms in vitro do not retain the spatial information present in the intact cell. We developed an in vivo assay to monitor the pattern of translation in the Drosophila oocyte, where several patterning determinants must be localized to specific regions. Using this assay, we showed that repression of translation by the Bruno protein is inhibited, and we could visualize when and where this occurs during oogenesis. Regional activation occurs not only at the site of mRNA localization, but more broadly in a graded fashion, and it does not require an activation element in the mRNA. We also show that Bruno dimerizes, and that dimerization is important for translational activation.
Zdroje
1. Martin KC, Ephrussi A (2009) mRNA localization: gene expression in the spatial dimension. Cell 136: 719–730. doi: 10.1016/j.cell.2009.01.044 19239891
2. Holt CE, Schuman EM (2013) The central dogma decentralized: new perspectives on RNA function and local translation in neurons. Neuron 80: 648–657. doi: 10.1016/j.neuron.2013.10.036 24183017
3. Jung H, Gkogkas CG, Sonenberg N, Holt CE (2014) Remote control of gene function by local translation. Cell 157: 26–40. doi: 10.1016/j.cell.2014.03.005 24679524
4. Lasko P (2012) mRNA localization and translational control in Drosophila oogenesis. Cold Spring Harb Perspect Biol 4.
5. St Johnston D (1995) The intracellular localization of messenger RNAs. Cell 81: 161–170. 7736568
6. Lehmann R, Nüsslein-Volhard C (1986) Abdominal segmentation, pole cell formation, and embryonic polarity require the localized activity of oskar, a maternal gene in Drosophila. Cell 47: 141–152. 3093084
7. Ephrussi A, Lehmann R (1992) Induction of germ cell formation by oskar. Nature 358: 387–392. 1641021
8. Smith JL, Wilson JE, Macdonald PM (1992) Overexpression of oskar directs ectopic activation of nanos and presumptive pole cell formation in Drosophila embryos. Cell 70: 849–859. 1516136
9. Ephrussi A, Dickinson LK, Lehmann R (1991) oskar organizes the germ plasm and directs localization of the posterior determinant nanos. Cell 66: 37–50. 2070417
10. Kim-Ha J, Smith JL, Macdonald PM (1991) oskar mRNA is localized to the posterior pole of the Drosophila ooctye. Cell 66: 23–35. 2070416
11. Kim-Ha J, Kerr K, Macdonald PM (1995) Translational regulation of oskar mRNA by bruno, an ovarian RNA-binding protein, is essential. Cell 81: 403–412. 7736592
12. Markussen F-H, Michon A-M, Breitwieser W, Ephrussi A (1995) Translational control of oskar generates Short OSK, the isoform that induces pole plasm assembly. Development 121: 3723–3732. 8582284
13. Rongo C, Gavis ER, Lehmann R (1995) Localization of oskar RNA regulates oskar translation and requires Oskar protein. Development 121: 2737–2746. 7555702
14. Lie Y, Macdonald PM (1999) Translational regulation of oskar mRNA occurs independent of the cap and poly(A) tail in Drosophila ovarian extracts. Development 126: 4989–4996. 10529417
15. Castagnetti S, Hentze MW, Ephrussi A, Gebauer F (2000) Control of oskar mRNA translation by Bruno in a novel cell-free system from Drosophila ovaries. Development 127: 1063–1068. 10662645
16. Nakamura A, Sato K, Hanyu-Nakamura K (2004) Drosophila Cup Is an eIF4E Binding Protein that Associates with Bruno and Regulates oskar mRNA Translation in Oogenesis. Dev Cell 6: 69–78. 14723848
17. Chekulaeva M, Hentze MW, Ephrussi A (2006) Bruno acts as a dual repressor of oskar translation, promoting mRNA oligomerization and formation of silencing particles. Cell 124: 521–533. 16469699
18. Jambor H, Brunel C, Ephrussi A (2011) Dimerization of oskar 3′ UTRs promotes hitchhiking for RNA localization in the Drosophila oocyte. RNA 17: 2049–2057. doi: 10.1261/rna.2686411 22028360
19. Besse F, Lopez de Quinto S, Marchand V, Trucco A, Ephrussi A (2009) Drosophila PTB promotes formation of high-order RNP particles and represses oskar translation. Genes Dev 23: 195–207. doi: 10.1101/gad.505709 19131435
20. Wilson JE, Connell JE, Macdonald PM (1996) aubergine enhances oskar translation in the Drosophila ovary. Development 122: 1631–1639. 8625849
21. Gunkel N, Yano T, Markussen FH, Olsen LC, Ephrussi A (1998) Localization-dependent translation requires a functional interaction between the 5′ and 3′ ends of oskar mRNA. Genes Dev 12: 1652–1664. 9620852
22. Chang JS, Tan L, Schedl P (1999) The Drosophila CPEB homolog, orb, is required for oskar protein expression in oocytes. Dev Biol 215: 91–106. 10525352
23. Micklem DR, Adams J, Grunert S, St Johnston D (2000) Distinct roles of two conserved Staufen domains in oskar mRNA localization and translation. EMBO J 19: 1366–1377. 10716936
24. Castagnetti S, Ephrussi A (2003) Orb and a long poly(A) tail are required for efficient oskar translation at the posterior pole of the Drosophila oocyte. Development 130: 835–843. 12538512
25. Munro TP, Kwon S, Schnapp BJ, St Johnston D (2006) A repeated IMP-binding motif controls oskar mRNA translation and anchoring independently of Drosophila melanogaster IMP. J Cell Biol 172: 577–588. 16476777
26. Reveal B, Yan N, Snee MJ, Pai CI, Gim Y, Macdonald PM (2010) BREs mediate both repression and activation of oskar mRNA translation and act in trans. Dev Cell 18: 496–502. doi: 10.1016/j.devcel.2009.12.021 20230756
27. Snee M, Benz D, Jen J, Macdonald PM (2008) Two distinct domains of Bruno bind specifically to the oskar mRNA. RNA Biol 5: 49–57.
28. Romaniuk PJ, Lowary P, Wu H-N, Stormo G, Uhlenbeck OC (1987) RNA binding site of R17 coat protein. Biochem 26: 1563–1568.
29. Kinoshita E, Kinoshita-Kikuta E, Takiyama K, Koike T (2006) Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Mol Cell Proteomics 5: 749–757. 16340016
30. Yoshida S, Müller HA, Wodarz A, Ephrussi A (2004) PKA-R1 spatially restricts Oskar expression for Drosophila embryonic patterning. Development 131: 1401–1410. 14993189
31. Flotow H, Graves PR, Wang AQ, Fiol CJ, Roeske RW, Roach PJ (1990) Phosphate groups as substrate determinants for casein kinase I action. J Biol Chem 265: 14264–14269. 2117608
32. White RR, Kwon YG, Taing M, Lawrence DS, Edelman AM (1998) Definition of optimal substrate recognition motifs of Ca2+-calmodulin-dependent protein kinases IV and II reveals shared and distinctive features. J Biol Chem 273: 3166–3172. 9452427
33. Zetterqvist O, Ragnarsson U, Engstrom L (1990) Substrate specificity of cyclic AMP-dependent protein kinase. Peptides and protein phosphorylation: 171–187.
34. LeCuyer K, Behlen L, Uhlenbeck O (1995) Mutants of the bacteriophage MS2 coat protein that alter its cooperative binding to RNA. Biochemistry 22: 10600–10606.
35. De Gregorio E, Preiss T, Hentze MW (1999) Translation driven by an eIF4G core domain in vivo. EMBO J 18: 4865–4874. 10469664
36. Vanzo NF, Ephrussi A (2002) Oskar anchoring restricts pole plasm formation to the posterior of the Drosophila oocyte. Development 129: 3705–3714. 12117819
37. Kim-Ha J, Webster PJ, Smith JL, Macdonald PM (1993) Multiple RNA regulatory elements mediate distinct steps in localization of oskar mRNA. Development 119: 169–178. 8275853
38. Ghosh S, Marchand V, Gáspár I, Ephrussi A (2012) Control of RNP motility and localization by a splicing-dependent structure in oskar mRNA. Nat Struct Mol Biol 19: 441–449. doi: 10.1038/nsmb.2257 22426546
39. Huang J, Zhou W, Dong W, Watson AM, Hong Y (2009) Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering. Proc Natl Acad Sci U S A 106: 8284–8289. doi: 10.1073/pnas.0900641106 19429710
40. Schüpbach T, Wieschaus E (1989) Female sterile mutations on the second chromosome of Drosophila melanogaster. I. Maternal effect mutations. Genetics 121: 101–117. 2492966
41. Snee MJ, Macdonald PM (2009) Dynamic organization and plasticity of sponge bodies. Dev Dyn 238: 918–930. doi: 10.1002/dvdy.21914 19301391
42. Webster PJ, Liang L, Berg CA, Lasko P, Macdonald PM (1997) Translational repressor bruno plays multiple roles in development and is widely conserved. Genes Dev 11: 2510–2521. 9334316
43. Reveal B, Garcia C, Ellington A, Macdonald PM (2011) Multiple RNA binding domains of Bruno confer recognition of diverse binding sites for translational repression. RNA Biol 8: 1047–1060. doi: 10.4161/rna.8.6.17542 21955496
44. Igreja C, Izaurralde E (2011) CUP promotes deadenylation and inhibits decapping of mRNA targets. Genes Dev 25: 1955–1967. doi: 10.1101/gad.17136311 21937713
45. Lane ME, Kalderon D (1994) RNA localization along the anteroposterior axis of the Drosophila oocyte requires PKA-mediated signal transduction to direct normal microtubule organization. Genes Dev 8: 2986–2995. 7528157
46. Huttelmaier S, Zenklusen D, Lederer M, Dictenberg J, Lorenz M, et al (2005) Spatial regulation of beta-actin translation by Src-dependent phosphorylation of ZBP1. Nature 438: 512–515. 16306994
47. Paquin N, Ménade M, Poirier G, Donato D, Drouet E, Chartrand P (2007) Local activation of yeast ASH1 mRNA translation through phosphorylation of Khd1p by the casein kinase Yck1p. Mol Cell 26: 795–809. 17588515
48. Deng Y, Singer RH, Gu W (2008) Translation of ASH1 mRNA is repressed by Puf6p-Fun12p/eIF5B interaction and released by CK2 phosphorylation. Genes Dev 22: 1037–1050. doi: 10.1101/gad.1611308 18413716
49. Saffman EE, Styhler S, Rother K, Li W, Richard S, Lasko P (1998) Premature translation of oskar in oocytes lacking the RNA-binding protein bicaudal-C. Mol Cell Biol 18: 4855–4862. 9671494
50. Nakamura A, Amikura R, Hanyu K, Kobayashi S (2001) Me31B silences translation of oocyte-localizing RNAs through the formation of cytoplasmic RNP complex during Drosophila oogenesis. Development 128: 3233–3242. 11546740
51. Wilhelm JE, Hilton M, Amos Q, Henzel WJ (2003) Cup is an eIF4E binding protein required for both the translational repression of oskar and the recruitment of Barentsz. J Cell Biol 163: 1197–1204. 14691132
52. Van Doren M, Williamson AL, Lehmann R (1998) Regulation of zygotic gene expression in Drosophila primordial germ cells. Curr Biol 8: 243–246. 9501989
53. Martin SG, St Johnston D (2003) A role for Drosophila LKB1 in anterior-posterior axis formation and epithelial polarity. Nature 421: 379–384. 12540903
54. Reich J, Snee MJ, Macdonald PM (2009) miRNA-dependent translational repression in the Drosophila ovary. PLoS One 4: e4669. doi: 10.1371/journal.pone.0004669 19252745
55. Haseloff J (1999) GFP variants for multispectral imaging of living cells. Methods Cell Biol 58: 139–151. 9891379
56. Verrotti AC, Wharton RP (2000) Nanos interacts with cup in the female germline of Drosophila. Development 127: 5225–5232. 11060247
57. Snee MJ, Macdonald PM (2004) Live imaging of nuage and polar granules: evidence against a precursor-product relationship and a novel role for Oskar in stabilization of polar granule components. J Cell Sci 117: 2109–2120. 15090597
58. Nelson MR, Leidal AM, Smibert CA (2004) Drosophila Cup is an eIF4E-binding protein that functions in Smaug-mediated translational repression. EMBO J 23: 150–159. 14685270
59. Lyon A, Reveal B, Macdonald PM, Hoffman D (2009) Bruno protein contains an expanded RNA recognition motif. Biochemistry 48: 12202–12212. doi: 10.1021/bi900624j 19919093
60. Filardo P, Ephrussi A (2003) Bruno regulates gurken during Drosophila oogenesis. Mech Dev 120: 289–297. 12591598
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2015 Číslo 2
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Genomic Selection and Association Mapping in Rice (): Effect of Trait Genetic Architecture, Training Population Composition, Marker Number and Statistical Model on Accuracy of Rice Genomic Selection in Elite, Tropical Rice Breeding Lines
- Discovery of Transcription Factors and Regulatory Regions Driving Tumor Development by ATAC-seq and FAIRE-seq Open Chromatin Profiling
- Evolutionary Signatures amongst Disease Genes Permit Novel Methods for Gene Prioritization and Construction of Informative Gene-Based Networks
- Proteotoxic Stress Induces Phosphorylation of p62/SQSTM1 by ULK1 to Regulate Selective Autophagic Clearance of Protein Aggregates