Nội dung được dịch bởi AI, chỉ mang tính chất tham khảo
Xác định các protein vận chuyển tiềm năng của protein transportin AtTRN1 trong Arabidopsis thaliana
Tóm tắt
Chúng tôi đã xác định được 23 protein mới có thể tương tác với AtTRN1. Những protein này là các ứng viên tiềm năng cho các protein vận chuyển của AtTRN1, giúp chúng tôi hiểu rõ hơn về chức năng của AtTRN1 trong Arabidopsis. Transportin 1 (TRN1) thực hiện việc vận chuyển protein qua lại giữa nhân và tế bào chất, đảm bảo rằng mỗi protein được chuyển đến ngăn đúng để thực hiện chức năng của nó. Những protein vận chuyển này tham gia vào nhiều quá trình quan trọng, chẳng hạn như cắt tỉa pre-mRNA thay thế, điều hòa phiên mã và dịch mã protein. Hiểu biết hiện tại về các protein vận chuyển do transportin 1 của Arabidopsis thaliana (AtTRN1) mang lại còn hạn chế. Ở đây, trước tiên chúng tôi đã áp dụng phương pháp sàng lọc hai hybrid nấm men (Y2H) để xác định các protein có thể tương tác với AtTRN1 trong Arabidopsis, và 12 protein mới đã được tìm thấy. Việc tìm kiếm motif PY-NLS trong 12 protein này cho thấy không có motif PY-NLS điển hình nào hiện diện. Chúng tôi sau đó đã điều tra các motif cụ thể sẽ trung gian các tương tác trong các chuỗi này, và phát hiện rằng mười ba mảnh đoạn rút gọn tương tác với AtTRN1, chứa 8 mảnh axit và 5 mảnh bazơ, tương ứng. Chúng tôi cũng đã tìm kiếm trong hệ proteome của Arabidopsis để tìm các homolog của các protein vận chuyển của nấm men Kapl04p và Kapβ2 ở động vật có vú, cùng với các protein chứa motif PY-NLS. Trong số những protein này, 11 protein đã được xác định là tương tác với AtTRN1. Các tương tác giữa cả 23 protein và AtTRN1 đã được xác nhận bởi cả hai phương pháp Y2H và bổ sung huỳnh quang phân tử hai (BiFC). Kết quả của chúng tôi cho thấy AtTRN1 nhận diện một phổ rộng các protein có chức năng đa dạng, có khả năng là các vật mang của AtTRN1. Nhìn chung, những kết quả này chứng minh tính khả thi và sức mạnh tiềm tàng của các phương pháp này trong việc xác định các protein vận chuyển của AtTRN1, và đại diện cho một bước đầu tiên quan trọng trong việc giải thích chức năng của AtTRN1.
Từ khóa
Tài liệu tham khảo
Aitchison JD, Blobel G, Rout MP (1996) Kap104p: a karyopherin involved in the nuclear transport of messenger RNA binding proteins. Science 274:624–627
Baake M, Bauerle M, Doenecke D, Albig W (2001) Core histones and linker histones are imported into the nucleus by different pathways. Eur J Cell Biol 80:669–677
Bairoch A, Boeckmann B, Ferro S, Gasteiger E (2004) Swiss-Prot: juggling between evolution and stability. Brief Bioinform 5:39–55
Barraud P, Banerjee S, Mohamed WI, Jantsch MF, Allain FH (2014) A bimodular nuclear localization signal assembled via an extended double-stranded RNA-binding domain acts as an RNA-sensing signal for transportin 1. Proc Natl Acad Sci USA 111:E1852–E1861
Blomster T, Salojarvi J, Sipari N, Brosche M, Ahlfors R, Keinanen M, Overmyer K, Kangasjarvi J (2011) Apoplastic reactive oxygen species transiently decrease auxin signaling and cause stress-induced morphogenic response in Arabidopsis. Plant Physiol 157:1866–1883
Bonifaci N, Moroianu J, Radu A, Blobel G (1997) Karyopherin beta2 mediates nuclear import of a mRNA binding protein. Proc Natl Acad Sci USA 94:5055–5060
Cansizoglu AE, Lee BJ, Zhang ZC, Fontoura BM, Chook YM (2007) Structure-based design of a pathway-specific nuclear import inhibitor. Nat Struct Mol Biol 14:452–454
Chang WL, Tarn WY (2009) A role for transportin in deposition of TTP to cytoplasmic RNA granules and mRNA decay. Nucleic Acids Res 37:6600–6612
Choi K, Kim J, Hwang HJ, Kim S, Park C, Kim SY, Lee I (2011) The FRIGIDA complex activates transcription of FLC, a strong flowering repressor in Arabidopsis, by recruiting chromatin modification factors. Plant Cell 23:289–303
Chook YM, Suel KE (2011) Nuclear import by karyopherin-betas: recognition and inhibition. Biochim Biophys Acta 1813:1593–1606
Cook A, Bono F, Jinek M, Conti E (2007) Structural biology of nucleocytoplasmic transport. Annu Rev Biochem 76:647–671
Datta S, Hettiarachchi GH, Deng XW, Holm M (2006) Arabidopsis CONSTANS-LIKE3 is a positive regulator of red light signaling and root growth. Plant Cell 18:70–84
Dean KA, von Ahsen O, Gorlich D, Fried HM (2001) Signal recognition particle protein 19 is imported into the nucleus by importin 8 (RanBP8) and transportin. J Cell Sci 114:3479–3485
Esteve-Bruna D, Perez-Perez JM, Ponce MR, Micol JL (2013) incurvata13, a novel allele of AUXIN RESISTANT6, reveals a specific role for auxin and the SCF complex in Arabidopsis embryogenesis, vascular specification, and leaf flatness. Plant Physiol 161:1303–1320
Favre N, Camps M, Arod C, Chabert C, Rommel C, Pasquali C (2008) Chemokine receptor CCR2 undergoes transportin1-dependent nuclear translocation. Proteomics 8:4560–4576
Fontoura BM, Blobel G, Yaseen NR (2000) The nucleoporin Nup98 is a site for GDP/GTP exchange on ran and termination of karyopherin beta 2-mediated nuclear import. J Biol Chem 275:31289–31296
Fried H, Kutay U (2003) Nucleocytoplasmic transport: taking an inventory. Cell Mol Life Sci 60:1659–1688
Gan ES, Xu Y, Wong JY, Goh JG, Sun B, Wee WY, Huang J, Ito T (2014) Jumonji demethylases moderate precocious flowering at elevated temperature via regulation of FLC in Arabidopsis. Nat Commun 5:5098
Gattiker A, Gasteiger E, Bairoch A (2002) ScanProsite: a reference implementation of a PROSITE scanning tool. Appl Bioinformatics 1:107–108
Gietz RD, Schiestl RH, Willems AR, Woods RA (1995) Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360
Gorlich D, Kutay U (1999) Transport between the cell nucleus and the cytoplasm. Annu Rev Cell Dev Biol 15:607–660
Gremski K, Ditta G, Yanofsky MF (2007) The HECATE genes regulate female reproductive tract development in Arabidopsis thaliana. Development 134:3593–3601
Guttinger S, Muhlhausser P, Koller-Eichhorn R, Brennecke J, Kutay U (2004) Transportin2 functions as importin and mediates nuclear import of HuR. Proc Natl Acad Sci USA 101:2918–2923
Han YJ, Cho KC, Hwang OJ, Choi YS, Shin AY, Hwang I, Kim JI (2012) Overexpression of an Arabidopsis beta-glucosidase gene enhances drought resistance with dwarf phenotype in creeping bentgrass. Plant Cell Rep 31:1677–1686
Horton P, Park KJ, Obayashi T, Fujita N, Harada H, Adams-Collier CJ, Nakai K (2007) WoLF PSORT: protein localization predictor. Nucleic Acids Res 35:W585–W587
Imasaki T, Shimizu T, Hashimoto H, Hidaka Y, Kose S, Imamoto N, Yamada M, Sato M (2007) Structural basis for substrate recognition and dissociation by human transportin 1. Mol Cell 28:57–67
Jakel S, Gorlich D (1998) Importin beta, transportin, RanBP5 and RanBP7 mediate nuclear import of ribosomal proteins in mammalian cells. EMBO J 17:4491–4502
Jones MA, Covington MF, DiTacchio L, Vollmers C, Panda S, Harmer SL (2010) Jumonji domain protein JMJD5 functions in both the plant and human circadian systems. Proc Natl Acad Sci USA 107:21623–21628
Lau CK, Delmar VA, Chan RC, Phung Q, Bernis C, Fichtman B, Rasala BA, Forbes DJ (2009) Transportin regulates major mitotic assembly events: from spindle to nuclear pore assembly. Mol Biol Cell 20:4043–4058
Lee BJ, Cansizoglu AE, Suel KE, Louis TH, Zhang Z, Chook YM (2006) Rules for nuclear localization sequence recognition by karyopherin beta 2. Cell 126:543–558
Lin YH, Huang LF, Hase T, Huang HE, Feng TY (2015) Expression of plant ferredoxin-like protein (PFLP) enhances tolerance to heat stress in Arabidopsis thaliana. N Biotechnol 32:235–242
Linding R, Jensen LJ, Diella F, Bork P, Gibson TJ, Russell RB (2003) Protein disorder prediction: implications for structural proteomics. Structure 11:1453–1459
Liu WX, Zhang FC, Zhang WZ, Song LF, Wu WH, Chen YF (2013) Arabidopsis Di19 functions as a transcription factor and modulates PR1, PR2, and PR5 expression in response to drought stress. Mol Plant 6:1487–1502
Lohr B, Streitner C, Steffen A, Lange T, Staiger D (2014) A glycine-rich RNA-binding protein affects gibberellin biosynthesis in Arabidopsis. Mol Biol Rep 41:439–445
Mahajan S, Sharma GK, Matura R, Subramaniam S, Mohapatra JK, Pattnaik B (2015) Construction and characterization of yeast two-hybrid cDNA library derived from LFBK cell line. Biologicals 43:202–208
Malnou CE, Salem T, Brockly F, Wodrich H, Piechaczyk M, Jariel-Encontre I (2007) Heterodimerization with Jun family members regulates c-Fos nucleocytoplasmic traffic. J Biol Chem 282:31046–31059
Merkle T (2003) Nucleo-cytoplasmic partitioning of proteins in plants: implications for the regulation of environmental and developmental signalling. Curr Genet 44:231–260
Merkle T, Grasser KD (2011) Unexpected mobility of plant chromatin-associated HMGB proteins. Plant Signal Behav 6:878–880
Muhlhausser P, Muller EC, Otto A, Kutay U (2001) Multiple pathways contribute to nuclear import of core histones. EMBO Rep 2:690–696
Nakaminami K, Hill K, Perry SE, Sentoku N, Long JA, Karlson DT (2009) Arabidopsis cold shock domain proteins: relationships to floral and silique development. J Exp Bot 60:1047–1062
Nakielny S, Siomi MC, Siomi H, Michael WM, Pollard V, Dreyfuss G (1996) Transportin: nuclear transport receptor of a novel nuclear protein import pathway. Exp Cell Res 229:261–266
Nakielny S, Shaikh S, Burke B, Dreyfuss G (1999) Nup153 is an M9-containing mobile nucleoporin with a novel Ran-binding domain. EMBO J 18:1982–1995
Ohn T, Kedersha N, Hickman T, Tisdale S, Anderson P (2008) A functional RNAi screen links O-GlcNAc modification of ribosomal proteins to stress granule and processing body assembly. Nat Cell Biol 10:1224–1231
Pollard VW, Michael WM, Nakielny S, Siomi MC, Wang F, Dreyfuss G (1996) A novel receptor-mediated nuclear protein import pathway. Cell 86:985–994
Robson F, Costa MM, Hepworth SR, Vizir I, Pineiro M, Reeves PH, Putterill J, Coupland G (2001) Functional importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants. Plant J 28:619–631
Scales JC, Parry MA, Salvucci ME (2014) A non-radioactive method for measuring Rubisco activase activity in the presence of variable ATP: ADP ratios, including modifications for measuring the activity and activation state of Rubisco. Photosynth Res 119:355–365
Schmal C, Reimann P, Staiger D (2013) A circadian clock-regulated toggle switch explains AtGRP7 and AtGRP8 oscillations in Arabidopsis thaliana. PLoS Comput Biol 9:e1002986
Schmidt F, Marnef A, Cheung MK, Wilson I, Hancock J, Staiger D, Ladomery M (2010) A proteomic analysis of oligo(dT)-bound mRNP containing oxidative stress-induced Arabidopsis thaliana RNA-binding proteins ATGRP7 and ATGRP8. Mol Biol Rep 37:839–845
Siomi H, Dreyfuss G (1995) A nuclear localization domain in the hnRNP A1 protein. J Cell Biol 129:551–560
Song HR, Song JD, Cho JN, Amasino RM, Noh B, Noh YS (2009) The RNA binding protein ELF9 directly reduces SUPPRESSOR OF OVEREXPRESSION OF CO1 transcript levels in arabidopsis, possibly via nonsense-mediated mRNA decay. Plant Cell 21:1195–1211
Stotz HU, Findling S, Nukarinen E, Weckwerth W, Mueller MJ, Berger S (2014) A tandem affinity purification tag of TGA2 for isolation of interacting proteins in Arabidopsis thaliana. Plant Signal Behav 9:e972794
Streitner C, Koster T, Simpson CG, Shaw P, Danisman S, Brown JW, Staiger D (2012) An hnRNP-like RNA-binding protein affects alternative splicing by in vivo interaction with transcripts in Arabidopsis thaliana. Nucleic Acids Res 40:11240–11255
Sun K, Cui Y, Hauser BA (2005) Environmental stress alters genes expression and induces ovule abortion: reactive oxygen species appear as ovules commit to abort. Planta 222:632–642
Tamura K, Hara-Nishimura I (2014) Functional insights of nucleocytoplasmic transport in plants. Front Plant Sci 5:118
Tillemans V, Leponce I, Rausin G, Dispa L, Motte P (2006) Insights into nuclear organization in plants as revealed by the dynamic distribution of Arabidopsis SR splicing factors. Plant Cell 18:3218–3234
To A, Joubes J, Barthole G, Lecureuil A, Scagnelli A, Jasinski S, Lepiniec L, Baud S (2012) WRINKLED transcription factors orchestrate tissue-specific regulation of fatty acid biosynthesis in Arabidopsis. Plant Cell 24:5007–5023
Twyffels L, Gueydan C, Kruys V (2014) Transportin-1 and Transportin-2: protein nuclear import and beyond. FEBS Lett 588:1857–1868
Waadt R, Schmidt LK, Lohse M, Hashimoto K, Bock R, Kudla J (2008) Multicolor bimolecular fluorescence complementation reveals simultaneous formation of alternative CBL/CIPK complexes in planta. Plant J 56:505–516
Weighardt F, Biamonti G, Riva S (1995) Nucleo-cytoplasmic distribution of human hnRNP proteins: a search for the targeting domains in hnRNP A1. J Cell Sci 108(Pt 2):545–555
Wirthmueller L, Roth C, Fabro G, Caillaud MC, Rallapalli G, Asai S, Sklenar J, Jones AM, Wiermer M, Jones JD, Banfield MJ (2015) Probing formation of cargo/importin-alpha transport complexes in plant cells using a pathogen effector. Plant J 81:40–52
Zhu W, Miao Q, Sun D, Yang G, Wu C, Huang J, Zheng C (2012) The mitochondrial phosphate transporters modulate plant responses to salt stress via affecting ATP and gibberellin metabolism in Arabidopsis thaliana. PLoS One 7:e43530
Ziemienowicz A, Haasen D, Staiger D, Merkle T (2003) Arabidopsis transportin1 is the nuclear import receptor for the circadian clock-regulated RNA-binding protein AtGRP7. Plant Mol Biol 53:201–212