Research Papers

Analysis of RNA Recognition and Binding Characteristics of OsCPPR1 Protein in Rice

Expand
  • State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources / Guangdong Laboratory for Lingnan Modern Agriculture / College of Life Sciences, South China Agricultural University, Guangzhou 510642, China
First author contact:

#These authors contributed equally to this work

Zhuang Chuxiong (zhuangcx@scau.edu.cn);

Received date: 2023-09-15

  Accepted date: 2023-11-30

  Online published: 2024-04-11

Abstract

Pentatricopeptide repeat (PPR) proteins represent one of the largest protein families in plants and typically localize to organelles like mitochondria and chloroplasts. By contrast, CYTOPLASM- LOCALIZED PPR1 (OsCPPR1) is a cytoplasm-localized PPR protein that can degrade OsGOLDEN- LIKE1 (OsGLK1) mRNA in the tapetum of rice anther. However, the mechanism, by which OsCPPR1 recognizes and binds to OsGLK1 transcripts, remains unknown. Through protein structure prediction and macromolecular docking experiments, we observed that distinct PPR motif structures of OsCPPR1 exhibited varying binding efficiencies to OsGLK1 RNA. Moreover, RNA-electrophoretic mobility shift assay experiment demonstrated that the recombinant OsCPPR1 can directly recognize and bind to OsGLK1 mRNA in vitro. This further confirmed that the mutations in the conserved amino acids in each PPR motif resulted in loss of activity, while truncation of OsCPPR1 decreased its binding efficiency. These findings collectively suggest that it may require some co-factors to assist in cleavage, a facet that warrants further exploration in subsequent studies.

Cite this article

Zheng Shaoyan, Chen Junyu, Li Huatian, Liu Zhenlan, Li Jing, Zhuang Chuxiong . Analysis of RNA Recognition and Binding Characteristics of OsCPPR1 Protein in Rice[J]. Rice Science, 2024 , 31(2) : 215 -225 . DOI: 10.1016/j.rsci.2023.11.011

References

[1] Abbas Y M, Pichlmair A, Górna M W, Superti-Furga G, Nagar B. 2013. Structural basis for viral 5′-PPP-RNA recognition by human IFIT proteins. Nature, 494: 60-64.
[2] Anandakrishnan R, Aguilar B, Onufriev A V. 2012. H++ 3.0: Automating pK prediction and the preparation of biomolecular structures for atomistic molecular modeling and simulations. Nucleic Acids Res, 40: W537-W541.
[3] Arenas-M A, González-Durán E, Gómez I, Burger M, Brennicke A, Takenaka M, Jordana X. 2018. The pentatricopeptide repeat protein MEF31 is required for editing at site 581 of the mitochondrial tatC transcript and indirectly influences editing at site 586 of the same transcript. Plant Cell Physiol, 59(2): 355-365.
[4] Arnal N, Quadrado M, Simon M, Mireau H. 2014. A restorer-of- fertility like pentatricopeptide repeat gene directs ribonucleolytic processing within the coding sequence of rps3-rpl16 and orf240a mitochondrial transcripts in Arabidopsis thaliana. Plant J, 78(1): 134-145.
[5] Ban T, Ke J Y, Chen R Z, Gu X, Tan M H, Zhou X E, Kang Y Y, Melcher K, Zhu J K, Xu H E. 2013. Structure of a PLS-class pentatricopeptide repeat protein provides insights into mechanism of RNA recognition. J Biol Chem, 288(44): 31540-31548.
[6] Barkan A, Small I. 2014. Pentatricopeptide repeat proteins in plants. Annu Rev Plant Biol, 65: 415-442.
[7] Barkan A, Rojas M, Fujii S, Yap A, Chong Y S, Bond C S, Small I. 2012. A combinatorial amino acid code for RNA recognition by pentatricopeptide repeat proteins. PLoS Genet, 8(8): e1002910.
[8] Beick S, Schmitz-Linneweber C, Williams-Carrier R, Jensen B, Barkan A. 2008. The pentatricopeptide repeat protein PPR5 stabilizes a specific tRNA precursor in maize chloroplasts. Mol Cell Biol, 28(17): 5337-5347.
[9] Biesiada M, Purzycka K J, Szachniuk M,. Blazewicz J, Adamiak R W. 2016. Automated RNA 3D structure prediction with RNAComposer. Methods Mol Biol, 1490: 199-215.
[10] Bryant N, Lloyd J, Sweeney C, Myouga F, Meinke D. 2011. Identification of nuclear genes encoding chloroplast-localized proteins required for embryo development in Arabidopsis. Plant Physiol, 155(4): 1678-1689.
[11] Chen X Z, Feng F, Qi W W, Xu L M, Yao D S, Wang Q, Song R T. 2017. Dek35 encodes a PPR protein that affects cis-splicing of mitochondrial nad4 intron 1 and seed development in maize. Mol Plant, 10(3): 427-441.
[12] Cheng S F, Gutmann B, Zhong X, Ye Y T, Fisher M F, Bai F Q, Castleden I, Song Y, Song B, Huang J Y, Liu X, Xu X, Lim B L, Bond C S, Yiu S M, Small I. 2016. Redefining the structural motifs that determine RNA binding and RNA editing by pentatricopeptide repeat proteins in land plants. Plant J, 85(4): 532-547.
[13] Dahan J, Mireau H. 2013. The Rf and Rf-like PPR in higher plants, a fast-evolving subclass of PPR genes. RNA Biol, 10(9): 1469-1476.
[14] de Longevialle A F, Hendrickson L, Taylor N L, Delannoy E, Lurin C, Badger M, Millar A H, Small I. 2008. The pentatricopeptide repeat gene OTP51 with two LAGLIDADG motifs is required for the cis-splicing of plastid ycf3 intron 2 in Arabidopsis thaliana. Plant J, 56(1): 157-168.
[15] des Francs-Small C C, Sanglard L V P, Small I. 2018. Targeted cleavage of nad6 mRNA induced by a modified pentatricopeptide repeat protein in plant mitochondria. Commun Biol, 1: 166.
[16] Ding Y H, Liu N Y, Tang Z S, Liu J, Yang W C. 2006. Arabidopsis GLUTAMINE-RICH PROTEIN23 is essential for early embryogenesis and encodes a novel nuclear PPR motif protein that interacts with RNA polymerase II subunit III. Plant Cell, 18(4): 815-830.
[17] Gruber A R, Lorenz R, Bernhart S H, Neuböck R, Hofacker I L. 2008. The Vienna RNA websuite. Nucleic Acids Res, 36: W70-W74.
[18] Gully B S, Cowieson N, Stanley W A, Shearston K, Small I D, Barkan A, Bond C S. 2015. The solution structure of the pentatricopeptide repeat protein PPR10 upon binding atpH RNA. Nucleic Acids Res, 43(3): 1918-1926.
[19] Hall T M. 2016. De-coding and re-coding RNA recognition by PUF and PPR repeat proteins. Curr Opin Struct Biol, 36: 116-121.
[20] Hammani K, Okuda K, Tanz S K, Chateigner-Boutin A L, Shikanai T, Small I. 2009. A study of new Arabidopsis chloroplast RNA editing mutants reveals general features of editing factors and their target sites. Plant Cell, 21(11): 3686-3699.
[21] Hammani K, Takenaka M, Miranda R, Barkan A. 2016. A PPR protein in the PLS subfamily stabilizes the 5′-end of processed rpl16 mRNAs in maize chloroplasts. Nucleic Acids Res, 44(9): 4278-4288.
[22] Hao Y Y, Wang Y L, Wu M M, Zhu X P, Teng X, Sun Y L, Zhu J P, Zhang Y Y, Jing R N, Lei J, Li J F, Bao X H, Wang C M, Wang Y H, Wan J M. 2019. The nuclear-localized PPR protein OsNPPR1 is important for mitochondrial function and endosperm development in rice. J Exp Bot, 70(18): 4705-4720.
[23] Hashimoto M, Endo T, Peltier G, Tasaka M, Shikanai T. 2003. A nucleus-encoded factor, CRR2, is essential for the expression of chloroplast ndhB in Arabidopsis. Plant J, 36( 4): 541-549.
[24] Hattori M, Miyake H, Sugita M. 2007. A pentatricopeptide repeat protein is required for RNA processing of clpP pre-mRNA in moss chloroplasts. J Biol Chem, 282(14): 10773-10782.
[25] Huang W F, Zhang Y, Shen L Q, Fang Q, Liu Q, Gong C B, Zhang C, Zhou Y, Mao C, Zhu Y L, Zhang J H, Chen H P, Zhang Y, Lin Y J, Bock R, Zhou F. 2020. Accumulation of the RNA polymerase subunit RpoB depends on RNA editing by OsPPR16 and affects chloroplast development during early leaf development in rice. New Phytol, 228(4): 1401-1416.
[26] Jiang H C, Lu Q, Qiu S Q, Yu H H, Wang Z J, Yu Z C, Lu Y R, Wang L, Xia F, Wu Y Y, Li F, Zhang Q L, Liu G, Song D D, Ma C L, Ding Q, Zhang X B, Zhang L, Zhang X T, Li X, Zhang J W, Xiao J H, Li X H, Wang N Y, Ouyang Y D, Zhou F S, Zhang Q F. 2022. Fujian cytoplasmic male sterility and the fertility restorer gene OsRf19 provide a promising breeding system for hybrid rice. Proc Natl Acad Sci USA, 119(34): e2208759119.
[27] Johnson X, Wostrikoff K, Finazzi G, Kuras R, Schwarz C, Bujaldon S, Nickelsen J, Stern D B, Wollman F A, Vallon O. 2010. MRL1, a conserved pentatricopeptide repeat protein, is required for stabilization of rbcL mRNA in Chlamydomonas and Arabidopsis. Plant Cell, 22(1): 234-248.
[28] Kazama T, Toriyama K. 2003. A pentatricopeptide repeat-containing gene that promotes the processing of aberrant atp6 RNA of cytoplasmic male-sterile rice. FEBS Lett, 544(1/3): 99-102.
[29] Kazama T, Nakamura T, Watanabe M, Sugita M, Toriyama K. 2008. Suppression mechanism of mitochondrial ORF79 accumulation by Rf1 protein in BT-type cytoplasmic male sterile rice. Plant J, 55(4): 619-628.
[30] Ke J Y, Chen R Z, Ban T, Zhou X E, Gu X, Eileen Tan M H, Chen C, Kang Y Y, Brunzelle J S, Zhu J K, Melcher K, Xu H E. 2013. Structural basis for RNA recognition by a dimeric PPR-protein complex. Nat Struct Mol Biol, 20(12): 1377-1382.
[31] Koussevitzky S, Nott A, Mockler T C, Hong F, Sachetto-Martins G, Surpin M, Lim J, Mittler R, Chory J. 2007. Signals from chloroplasts converge to regulate nuclear gene expression. Science, 316: 715-719.
[32] Li X J, Zhang Y F, Hou M M, Sun F, Shen Y, Xiu Z H, Wang X M, Chen Z L, Sun S S M, Small I, Tan B C. 2014. Small kernel 1encodes a pentatricopeptide repeat protein required for mitochondrial nad7 transcript editing and seed development in maize (Zea mays) and rice (Oryza sativa). Plant J, 79(5): 797-809.
[33] Liu Y J, Xiu Z H, Meeley R, Tan B C. 2013. Empty pericarp5 encodes a pentatricopeptide repeat protein that is required for mitochondrial RNA editing and seed development in maize. Plant Cell, 25(3): 868-883.
[34] Lurin C, Andrés C, Aubourg S, Bellaoui M, Bitton F, Bruyère C, Caboche M, Debast C, Gualberto J, Hoffmann B, Lecharny A, Le Ret M, Martin-Magniette M L, Mireau H, Peeters N, Renou J P, Szurek B, Taconnat L, Small I. 2004. Genome-wide analysis of Arabidopsis pentatricopeptide repeat proteins reveals their essential role in organelle biogenesis. Plant Cell, 16(8): 2089-2103.
[35] Ma F, Hu Y C, Ju Y, Jiang Q R, Cheng Z J, Zhang Q,Sodmergen. 2017. A novel tetratricopeptide repeat protein, WHITE TO GREEN1, is required for early chloroplast development and affects RNA editing in chloroplasts. J Exp Bot, 68(21/22): 5829-5843.
[36] Müller-McNicoll M, Rossbach O, Hui J Y, Medenbach J. 2019. Auto-regulatory feedback by RNA-binding proteins. J Mol Cell Biol, 11(10): 930-939.
[37] O’Toole N, Hattori M, Andres C, Iida K, Lurin C, Schmitz- Linneweber C, Sugita M, Small I. 2008. On the expansion of the pentatricopeptide repeat gene family in plants. Mol Biol Evol, 25(6): 1120-1128.
[38] Pettersen E F, Goddard T D, Huang C C, Couch G S, Greenblatt D M, Meng E C, Ferrin T E. 2004. UCSF Chimera: A visualization system for exploratory research and analysis. J Comput Chem, 25(13): 1605-1612.
[39] Pfalz J, Liere K, Kandlbinder A, Dietz K J, Oelmüller R. 2006. pTAC2, -6, and -12 are components of the transcriptionally active plastid chromosome that are required for plastid gene expression. Plant Cell, 18(1): 176-197.
[40] Prikryl J, Rojas M, Schuster G, Barkan A. 2011. Mechanism of RNA stabilization and translational activation by a pentatricopeptide repeat protein. Proc Natl Acad Sci USA, 108(1): 415-420.
[41] Rovira A G, Smith A G. 2019. PPR proteins-orchestrators of organelle RNA metabolism. Physiol Plant, 166(1): 451-459.
[42] Saha D, Prasad A M, Srinivasan R. 2007. Pentatricopeptide repeat proteins and their emerging roles in plants. Plant Physiol Biochem, 45(8): 521-534.
[43] Schmitz-Linneweber C, Williams-Carrier R, Barkan A. 2005. RNA immunoprecipitation and microarray analysis show a chloroplast pentatricopeptide repeat protein to be associated with the 5′ region of mRNAs whose translation it activates. Plant Cell, 17(10): 2791-2804.
[44] Shen C C, Zhang D L, Guan Z Y, Liu Y X, Yang Z, Yang Y, Wang X, Wang Q, Zhang Q X, Fan S L, Zou T T, Yin P. 2016. Structural basis for specific single-stranded RNA recognition by designer pentatricopeptide repeat proteins. Nat Commun, 7: 11285.
[45] Small I D, Peeters N. 2000. The PPR motif: A TPR-related motif prevalent in plant organellar proteins. Trends Biochem Sci, 25(2): 45-47.
[46] Sota F, Ian S. 2011. The evolution of RNA editing and pentatricopeptide repeat genes. New Phytol, 191(1): 37-47.
[47] Stern D B, Goldschmidt-Clermont M, Hanson M R. 2010. Chloroplast RNA metabolism. Annu Rev Plant Biol, 61: 125-155.
[48] Tavares-Carreón F, Camacho-Villasana Y, Zamudio-Ochoa A, Shingú-Vázquez M, Torres-Larios A, Pérez-Martínez X. 2008. The pentatricopeptide repeats present in Pet309 are necessary for translation but not for stability of the mitochondrial COX1 mRNA in yeast. J Biol Chem, 283(3): 1472-1479.
[49] Wang Z H, Zou Y J, Li X Y, Zhang Q Y, Chen L T, Wu H, Su D H, Chen Y L, Guo J X, Luo D, Long Y M, Zhong Y, Liu Y G. 2006. Cytoplasmic male sterility of rice with boro II cytoplasm is caused by a cytotoxic peptide and is restored by two related PPR motif genes via distinct modes of mRNA silencing. Plant Cell, 18(3): 676-687.
[50] Waters M T, Wang P, Korkaric M, Capper R G, Saunders N J, Langdale J A. 2009. GLK transcription factors coordinate expression of the photosynthetic apparatus in Arabidopsis. Plant Cell, 21(4): 1109-1128.
[51] Wu L L, Wu J, Liu Y X, Gong X D, Xu J L, Lin D Z, Dong Y J. 2016. The rice pentatricopeptide repeat gene TCD10 is needed for chloroplast development under cold stress. Rice, 9(1): 67.
[52] Xie T T, Chen D, Wu J, Huang X R, Wang Y F, Tang K L, Li J Y, Sun M X, Peng X B. 2016. Growing Slowly 1 locus encodes a PLS-type PPR protein required for RNA editing and plant development in Arabidopsis. J Exp Bot, 67(19): 5687-5698.
[53] Yan Y M, Zhang D, Zhou P, Li B T, Huang S Y. 2017. HDOCK: A web server for protein-protein and protein-DNA/RNA docking based on a hybrid strategy. Nucleic Acids Res, 45: W365-W373.
[54] Yang H, Wang Y L, Tian Y L, Teng X, Lv Z H, Lei J, Duan E C, Dong H, Yang X, Zhang Y Y, Sun Y L, Chen X L, Bao X H, Chen R B, Gu C W, Zhang Y P, Jiang X K, Ma W Y, Zhang P C, Ji Y, Zhang Y, Wang Y H, Wan J M. 2023. Rice FLOURY ENDOSPERM22, encoding a pentatricopeptide repeat protein, is involved in both mitochondrial RNA splicing and editing and is crucial for endosperm development. J Integr Plant Biol, 65(3): 755-771.
[55] Yang J Y, Zhang M, Wang X. 2018. Crystal structure of the chloroplast RNA editing factor MORF2. Biochem Biophys Res Commun, 495(2): 2038-2043.
[56] Yap A, Kindgren P, Colas des Francs-Small C, Kazama T, Tanz S K, Toriyama K, Small I. 2015. AEF1/MPR25 is implicated in RNA editing of plastid atpF and mitochondrial nad5, and also promotes atpF splicing in Arabidopsis and rice. Plant J, 81(5): 661-669.
[57] Yin P, Li Q X, Yan C Y, Liu Y, Liu J J, Yu F, Wang Z, Long J F, He J H, Wang H W, Wang J W, Zhu J K, Shi Y G, Yan N E. 2013. Structural basis for the modular recognition of single-stranded RNA by PPR proteins. Nature, 504: 168-171.
[58] Zhang Y, Lu C M. 2019. The enigmatic roles of PPR-SMR proteins in plants. Adv Sci, 6(13): 1900361.
[59] Zheng S Y, Li J, Ma L, Wang H L, Zhou H, Ni E D, Jiang D G, Liu Z L, Zhuang C X. 2019. OsAGO2 controls ROS production and the initiation of tapetal PCD by epigenetically regulating OsHXK1 expression in rice anthers. Proc Natl Acad Sci USA, 116(15): 7549-7558.
[60] Zheng S Y, Dong J F, Lu J Q, Li J, Jiang D G, Yu H P, Ye S M, Bu W L, Liu Z L, Zhou H, Ding Y L, Zhuang C X. 2022. A cytosolic pentatricopeptide repeat protein is essential for tapetal plastid development by regulating OsGLK1 transcript levels in rice. New Phytol, 234(5): 1678-1695.
[61] Zhou W, Lu Q T, Li Q W, Wang L, Ding S H, Zhang A H, Wen X G, Zhang L X, Lu C M. 2017. PPR-SMR protein SOT1 has RNA endonuclease activity. Proc Natl Acad Sci USA, 114(8): E1554-E1563.
[62] Zoschke R, Kroeger T, Belcher S, Schöttler M A, Barkan A, Schmitz-Linneweber C. 2012. The pentatricopeptide repeat- SMR protein ATP4 promotes translation of the chloroplast atpB/E mRNA. Plant J, 72(4): 547-558.
[63] Zsigmond L, Szepesi Á, Tari I, Rigó G, Király A, Szabados L. 2012. Overexpression of the mitochondrial PPR40 gene improves salt tolerance in Arabidopsis. Plant Sci, 182: 87-93.
Outlines

/

浙ICP备05004719号-15   公安备案号:33010302003355
Copyright © Editorial office of Rice Science
Tel: 0571-63371017 E-mail: crrn@fy.hz.zn.cn; cjrs278@gmail.com
Supported by Beijing Magtech Co., Ltd.
Total visitors:  Visitors of today:  Now online: