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Molecular Cancer
Published in: Molecular Cancer 1/2014

Open Access 01-12-2014 | Research

Long-term reduction of T-cell intracellular antigens leads to increased beta-actin expression

Authors: Isabel Carrascoso, Carmen Sánchez-Jiménez, José M Izquierdo

Published in: Molecular Cancer | Issue 1/2014

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Abstract

Background

The permanent down-regulated expression of T-cell intracellular antigen (TIA) proteins in HeLa cells improves cytoskeleton-mediated functions such as cell proliferation and tumor growth.

Methods

Making use of human and mouse cells with knocked down/out expression of T-cell intracellular antigen 1 (TIA1) and/or TIA1 related/like (TIAR/TIAL1) proteins and classical RNA (e.g. reverse transcription-quantitative polymerase chain reaction, polysomal profiling analysis using sucrose gradients, immunoblotting, immunoprecipitation, electrophoretic mobility shift assays, ultraviolet light crosslinking and poly (A+) test analysis) and cellular (e.g. immunofluorescence microscopy and quimeric mRNA transfections) biology methods, we have analyzed the regulatory role of TIA proteins in the post-transcriptional modulation of beta-actin (ACTB) mRNA.

Results

Our observations show that the acquisition of above cellular capacities is concomitant with increased expression levels of the actin beta subunit (ACTB) protein. Regulating TIA abundance does not modify ACTB mRNA levels, however, an increase of ACTB mRNA translation is observed. This regulatory capacity of TIA proteins is linked to the ACTB mRNA 3′-untranslated region (3′-UTR), where these proteins could function as RNA binding proteins. The expression of GFP from a chimeric reporter containing human ΑCΤΒ 3′-UTR recapitulates the translational control found by the endogenous ACTB mRNA in the absence of TIA proteins. Additionally, murine embryonic fibroblasts (MEF) knocked out for TIA1 rise mouse ACTB protein expression compared to the controls. Once again steady-state levels of mouse ACTB mRNA remained unchanged.

Conclusions

Collectively, these results suggest that TIA proteins can function as long-term regulators of the ACTB mRNA metabolism in mouse and human cells.
Appendix
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Literature
1.
Piecyk M, Wax S, Beck AR, Kedersha N, Gupta M, Maritim B, Chen S, Gueydan C, Kruys V, Streuli M, Anderson P: TIA1 is a translational silencer that selectively regulates the expression of TNF-α. EMBO J. 2000, 19: 4154-4163. 10.1093/emboj/19.15.4154PubMedCentralCrossRefPubMed
2.
Beck AR, Miller JJ, Anderson P, Streuli M: RNA-binding protein TIAR is essential for primordial germ cell development. Proc Natl Acad Sci. 1998, 95: 2331-2336. 10.1073/pnas.95.5.2331PubMedCentralCrossRefPubMed
3.
Kharraz Y, Salmand PA, Camus A, Auriol J, Gueydan C, Kruys V, Morello D: Impaired embryonic development in mice overexpressing the RNA-binding protein TIAR. PLoS One. 2010, 5: e11352- 10.1371/journal.pone.0011352PubMedCentralCrossRefPubMed
4.
Suswam EA, Li YY, Mahtani H, King PH: Novel DNA-binding properties of the RNA-binding protein TIAR. Nucleic Acids Res. 2005, 33: 4507-4518. 10.1093/nar/gki763PubMedCentralCrossRefPubMed
5.
Das R, Yu J, Zhang Z, Gygi MP, Krainer AR, Gygi SP, Reed R: SR proteins function in coupling RNAP II transcription to pre-mRNA splicing. Mol Cell. 2007, 26: 867-881. 10.1016/j.molcel.2007.05.036CrossRefPubMed
6.
McAlinden A, Liang L, Mukudai Y, Imamura T, Sandell LJ: Nuclear protein TIA1 regulates COL2A1 alternative splicing and interacts with precursor mRNA and genomic DNA. J Biol Chem. 2007, 282: 24444-24454. 10.1074/jbc.M702717200CrossRefPubMed
7.
Kim HS, Kuwano Y, Zhan M, Pullmann R, Mazan-Mamczarz K, Li H, Kedersha N, Anderson P, Wilce MC, Gorospe M, Wilce JA: Elucidation of a C-rich signature motif in target mRNAs of RNA-binding protein TIAR. Mol Cell Biol. 2007, 27: 6806-6817. 10.1128/MCB.01036-07PubMedCentralCrossRefPubMed
8.
Reyes R, Alcalde J, Izquierdo JM: Depletion of T-cell intracellular antigen (TIA)-proteins promotes cell proliferation. Genome Biol. 2009, 10: R87- 10.1186/gb-2009-10-8-r87PubMedCentralCrossRefPubMed
9.
Wang Z, Kayikci M, Briese M, Zarnack K, Luscombe NM, Rot G, Zupan B, Curk T, Ule J: iCLIP predicts the dual splicing effects of TIA-RNA interactions. PLoS Biol. 2010, 8: e1000530- 10.1371/journal.pbio.1000530PubMedCentralCrossRefPubMed
10.
Del Gatto-Konczak F, Bourgeois CF, Le Guiner C, Kister L, Gesnel MC, Stévenin J, Breathnach R: The RNA-binding protein TIA1 is a novel mammalian splicing regulator acting through intron sequences adjacent to a 5′ splice site. Mol Cell Biol. 2000, 20: 6287-6299. 10.1128/MCB.20.17.6287-6299.2000PubMedCentralCrossRefPubMed
11.
Förch P, Puig O, Kedersha N, Martínez C, Granneman S, Séraphin B, Anderson P, Valcárcel J: The apoptosis-promoting factor TIA1 is a regulator of alternative pre-mRNA splicing. Mol Cell. 2000, 6: 1089-1098. 10.1016/S1097-2765(00)00107-6CrossRefPubMed
12.
Förch P, Puig O, Martínez C, Séraphin B, Valcárcel J: The splicing regulator TIA1 interacts with U1-C to promote U1 snRNP recruitment to 5′ splice sites. EMBO J. 2002, 21: 6882-6892. 10.1093/emboj/cdf668PubMedCentralCrossRefPubMed
13.
Izquierdo JM, Majós N, Bonnal S, Martínez C, Castelo R, Guigó R, Bilbao D, Valcárcel J: Regulation of Fas alternative splicing by antagonistic effects of TIA1 and PTB on exon definition. Mol Cell. 2005, 19: 475-484. 10.1016/j.molcel.2005.06.015CrossRefPubMed
14.
Aznarez I, Barash Y, Shai O, He D, Zielenski J, Tsui LC, Parkinson J, Frey BJ, Rommens JM, Blencowe BJ: A systematic analysis of intronic sequences downstream of 5′ splice sites reveals a widespread role for U-rich motifs and TIA1/TIAL1 proteins in alternative splicing regulation. Genome Res. 2008, 18: 1247-1258. 10.1101/gr.073155.107PubMedCentralCrossRefPubMed
15.
Gueydan C, Droogmans L, Chalon P, Huez G, Caput D, Kruys V: Identification of TIAR as a protein binding to the translational regulatory AU-rich element of tumor necrosis factor α mRNA. J Biol Chem. 1999, 274: 2322-2326. 10.1074/jbc.274.4.2322CrossRefPubMed
16.
Phillips K, Kedersha N, Shen L, Blackshear PJ, Anderson P: Arthritis suppressor genes TIA1 and TTP dampen the expression of tumor necrosis factor α, cyclooxygenase 2 and inflammatory arthritis. Proc Natl Acad Sci. 2004, 101: 2011-2016. 10.1073/pnas.0400148101PubMedCentralCrossRefPubMed
17.
Yamasaki S, Stoecklin G, Kedersha N, Simarro M, Anderson P: T-cell intracellular antigen-1 (TIA1)-induced translational silencing promotes the decay of selected mRNAs. J Biol Chem. 2007, 282: 30070-30077. 10.1074/jbc.M706273200CrossRefPubMed
18.
Dember LM, Kim ND, Liu KQ, Anderson P: Individual RNA recognition motifs of TIA1 and TIAR have different RNA binding specificities. J Biol Chem. 1996, 271: 2783-2788. 10.1074/jbc.271.5.2783CrossRefPubMed
19.
López De Silanes I, Galbán S, Martindale JL, Yang X, Mazan-Mamczarz K, Indig FE, Falco G, Zhan M, Gorospe M: Identification and functional outcome of mRNAs associated with RNA-binding protein TIA1. Mol Cell Biol. 2005, 25: 9520-9531. 10.1128/MCB.25.21.9520-9531.2005PubMedCentralCrossRefPubMed
20.
Mazan-Mamczarz K, Lal A, Martindale JL, Kawai T, Gorospe M: Translational repression by RNA-binding protein TIAR. Mol Cell Biol. 2006, 26: 2716-2727. 10.1128/MCB.26.7.2716-2727.2006PubMedCentralCrossRefPubMed
21.
22.
Thoreen CC, Chantranupong L, Keys HR, Wang T, Grays NS, Sabatini DM: A unifying model for mTORC1-mediated regulation of mRNA translation. Nature. 2012, 485: 109-113. 10.1038/nature11083PubMedCentralCrossRefPubMed
23.
Tian Q, Streuli M, Saito H, Schlossman SF, Anderson P: A polyadenylate binding protein localized to the granules of cytolytic lymphocytes induces DNA fragmentation in target cells. Cell. 1991, 67: 629-639. 10.1016/0092-8674(91)90536-8CrossRefPubMed
24.
Kawakami A, Tian Q, Duan X, Streuli M, Schlossman SF, Anderson P: Identification and functional characterization of a TIA1-related nucleolysin. Proc Natl Acad Sci. 1992, 89: 8681-8685. 10.1073/pnas.89.18.8681PubMedCentralCrossRefPubMed
25.
Iseni F, Garcin D, Nishio M, Kedersha N, Anderson P, Kolakofsky D: Sendai virus trailer RNA binds TIAR, a cellular protein involved in virus-induced apoptosis. EMBO J. 2002, 21: 5141-5150. 10.1093/emboj/cdf513PubMedCentralCrossRefPubMed
26.
Li W, Li Y, Kedersha N, Anderson P, Emara M, Swiderek KM, Moreno GT, Brinton MA: Cell proteins TIA1 and TIAR interact with the 3′ stem-loop of the West Nile virus complementary minus-strand RNA and facilitate virus replication. J Virol. 2002, 76: 11989-20000. 10.1128/JVI.76.23.11989-12000.2002PubMedCentralCrossRefPubMed
27.
Izquierdo JM, Alcalde J, Carrascoso I, Reyes R, Ludeña MD: Knockdown of T-cell intracellular antigens triggers cell proliferation, invasion and tumor growth. Biochem J. 2011, 435: 337-344. 10.1042/BJ20101030CrossRefPubMed
28.
Yamazaki D, Kurisu S, Takenawa T: Regulation of cancer cell motility through actin reorganization. Cancer Sci. 2005, 906: 379-386.CrossRef
29.
Dormoy-Raclet V, Ménard I, Clair E, Kurban G, Mazroui R, Di Marco S, Von Roretz C, Pause A, Gallouzi IE: The RNA-binding protein HuR promotes cell migration and cell invasion by stabilizing the beta-actin mRNA in a U-rich-element-dependent manner. Mol Cell Biol. 2007, 27: 5365-5380. 10.1128/MCB.00113-07PubMedCentralCrossRefPubMed
30.
Vandekerckhove J, Weber K: At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 1978, 126: 783-802. 10.1016/0022-2836(78)90020-7CrossRefPubMed
31.
Olave IA, Reck-Peterson S, Crabtree GR: Nuclear actin and actin-related proteins in chromatin remodelling. Annu Rev Biochem. 2002, 71: 755-781. 10.1146/annurev.biochem.71.110601.135507CrossRefPubMed
32.
Kislauskis EH, Li Z, Singer RH, Taneja KL: Isoform-specific 3′-untranslated sequences sort alpha-cardiac and beta-cytoplasmic actin messenger RNAs to different cytoplasmic compartments. J Cell Biol. 1993, 123: 165-172. 10.1083/jcb.123.1.165CrossRefPubMed
33.
Chartrand P, Singer RH, Long RM: RNP localization and transport in yeast. Annu Rev Cell Dev Biol. 2001, 17: 297-310.CrossRefPubMed
34.
35.
Buxbaum AR, Wu B, Singer RH: Single β-actin mRNA detection in neurons reveals a mechanism for regulating its translatability. Science. 2014, 343: 419-422. 10.1126/science.1242939PubMedCentralCrossRefPubMed
36.
37.
Hüttelmaier S, Zenklusen D, Lederer M, Dictenberg J, Lorenz M, Meng X, Bassell GJ, Condelis J, Singer RH: Spatial regulation of beta-actin translation by Src-dependent phosphorylation of ZBP1. Nature. 2005, 438: 512-515. 10.1038/nature04115CrossRefPubMed
38.
Chao JA, Patskovsky Y, Patel V, Levy M, Almo SC, Singer RH: ZBP1 recognition of beta-actin zipcode induces RNA looping. Genes Dev. 2010, 24: 148-158. 10.1101/gad.1862910PubMedCentralCrossRefPubMed
39.
Patel VL, Mitra S, Harris R, Buxbaum AR, Lionnet T, Brenowitz M, Girvin M, Levy M, Almo SC, Singer RH, Chao JA: Spatial arrangement of an RNA zipcode identifies mRNAs under post-transcriptional control. Genes Dev. 2012, 26: 43-53. 10.1101/gad.177428.111PubMedCentralCrossRefPubMed
40.
Welch MD, Mullins RD: Cellular control of actin nucleation. Annu Rev Cell Dev Biol. 2002, 18: 247-288. 10.1146/annurev.cellbio.18.040202.112133CrossRefPubMed
41.
Stöhr N, Köhn M, Lederer M, Glass M, Reinke C, Singer RH, Hüttelmaier S: IGF2BP1 promotes cell migration by regulatin MK5 and PTEN signalling. Genes Dev. 2012, 26: 176-189. 10.1101/gad.177642.111PubMedCentralCrossRefPubMed
42.
Katz ZB, Wells AL, Park HY, Wu B, Shenoy SM, Singer RH: β-actin mRNA compartmentalization enhances focal adhesion stability and directs cell migration. Genes Dev. 2012, 26: 1885-1890. 10.1101/gad.190413.112PubMedCentralCrossRefPubMed
43.
Kedersha N, Anderson P: Stress granules: sites of mRNA triage that regulate mRNA stability and translatability. Biochem Soc Trans. 2002, 30: 963-969.CrossRefPubMed
44.
Izquierdo JM: Cell-specific regulation of Fas exon 6 splicing mediated by Hu antigen R. Biochem Biophys Res Commun. 2010, 402: 324-328. 10.1016/j.bbrc.2010.10.025CrossRefPubMed
45.
Izquierdo JM, Valcárcel J: Two isoforms of the T-cell intracellular antigen 1 (TIA1) splicing factors display distinct splicing regulation activities. Control of TIA1 isoform ratio by TIA1 related protein. J Biol Chem. 2007, 282: 19410-19417. 10.1074/jbc.M700688200CrossRefPubMed
46.
Izquierdo JM: Hu antigen R (HuR) functions as an alternative pre-mRNA splicing regulator of Fas apoptosis-promoting receptor on exon definition. J Biol Chem. 2008, 283: 19077-19084. 10.1074/jbc.M800017200CrossRefPubMed
47.
Izquierdo JM: Fas splicing regulation during early apoptosis is linked to caspase-mediated cleavage of U2AF65. Mol Biol Cell. 2008, 19: 3299-3307. 10.1091/mbc.E07-11-1125PubMedCentralCrossRefPubMed
48.
Lejeune F, Maquat LE: Immunopurification and analysis of protein and RNA components of mRNP in mammalian cells. Methods in Molecular Biology: mRNA Processing and Metabolism. Edited by: Schoenberg DR. 2004, 115-124. Totowa, New Jersey: Humana Press,CrossRef
49.
Sallés FJ, Richards WG, Strickland S: Assaying the polyadenylation state of mRNAs. Methods Enzymol. 1999, 17: 38-45. 10.1006/meth.1998.0705.CrossRef
Metadata
Title
Long-term reduction of T-cell intracellular antigens leads to increased beta-actin expression
Authors
Isabel Carrascoso
Carmen Sánchez-Jiménez
José M Izquierdo
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2014
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-13-90

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