Abstract
The polymorphic S-locus regulating self-incompatibility (SI) in Petunia contains the S-RNase gene and a number of S-locus F-box (SLF) genes. While penetrating the style through the stigma, a pollen tube takes up all S-RNases, but only self S-RNase inhibits pollen tube growth. Recent evidence suggests that SLFs produced by pollen collectively interact with and detoxify non-self S-RNases, but none can interact with self S-RNase. An SLF may be the F-box protein component of an SCF complex (containing Cullin1, Skp1 and Rbx1), which mediates ubiquitination of protein substrates for degradation by the 26S proteasome. However, the precise nature of the complex is unknown. We used pollen extracts of a transgenic plant over-expressing GFP-fused S2-SLF1 (SLF1 of S 2-haplotype) for co-immunoprecipitation (Co-IP) followed by mass spectrometry (MS). We identified PiCUL1-P (a pollen-specific Cullin1), PiSSK1 (a pollen-specific Skp1-like protein) and PiRBX1 (an Rbx1). To validate the results, we raised transgenic plants over-expressing PiSSK1:FLAG:GFP and used pollen extracts for Co-IP–MS. The results confirmed the presence of PiCUL1-P and PiRBX1 in the complex and identified two different SLFs as the F-box protein component. Thus, all but Rbx1 of the complex may have evolved in SI, and all SLFs may be the F-box component of similar complexes.
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References
Ai YJ, Singh A, Coleman CE, Ioerger TR, Kheyrpour A, Kao T-h (1990) Self-incompatibility in Petunia inflata: isolation and characterization of cDNAs encoding 3 S-allele-associated proteins. Sex Plant Reprod 3(2):130–138
Bai C, Sen P, Hofmann K, Ma L, Goebl M, Harper JW, Elledge SJ (1996) SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box. Cell 86(2):263–274
Chen X, Simon ES, Xiang Y, Kachman M, Andrews PC, Wang Y (2010) Quantitative proteomics analysis of cell cycle-regulated Golgi disassembly and reassembly. J Biol Chem 285(10):7197–7207
de Nettancourt D (2001) Incompatibility and incongruity in wild and cultivated plants. Springer, Berlin
Entani T, Iwano M, Shiba H, Che FS, Isogai A, Takayama S (2003) Comparative analysis of the self-incompatibility (S-) locus region of Prunus mume: identification of a pollen-expressed F-box gene with allelic diversity. Genes Cells 8(3):203–213
Goldraij A, Kondo K, Lee CB, Hancock CN, Sivaguru M, Vazquez-Santana S, Kim S, Phillips TE, Cruz-Garcia F, McClure B (2006) Compartmentalization of S-RNase and HT-B degradation in self incompatible Nicotiana. Nature 439(7078):805–810
Heginbotham L, Odessey E, Miller C (1997) Tetrameric stoichiometry of a prokaryotic K+ channel. Biochemistry 36(33):10335–10342
Hershko A, Heller H, Elias S, Ciechanover A (1983) Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown. J Biol Chem 258(13):8206–8214
Hua ZH, Kao T-h (2006) Identification and characterization of components of a putative Petunia S-locus F-box-containing E3 ligase complex involved in S-RNase-based self-incompatibility. Plant Cell 18(10):2531–2553
Hua Z, Kao T-h (2008) Identification of major lysine residues of S3-RNase of Petunia inflata involved in ubiquitin-26S proteasome-mediated degradation in vitro. Plant J 54(6):1094–1104
Hua ZH, Meng XY, Kao T-h (2007) Comparison of Petunia inflata S-locus F-box protein (Pi SLF) with Pi SLF-like proteins reveals its unique function in S-RNase-based self-incompatibility. Plant Cell 19(11):3593–3609
Huang S, Lee HS, Karunanandaa B, Kao T-h (1994) Ribonuclease activity of Petunia inflata S-Proteins is essential for rejection of self-pollen. Plant Cell 6(7):1021–1028
Huang J, Zhao L, Yang QY, Xue YB (2006) AhSSK1, a novel SKP1-like protein that interacts with the S-locus F-box protein SLF. Plant J 46(5):780–793
Ikeda K, Igic B, Ushijima K, Yamane H, Hauck NR, Nakano R, Sassa H, Iezzoni AF, Kohn JR, Tao R (2004) Primary structural features of the S haplotype-specific F-box protein, SFB in Prunus. Sex Plant Reprod 16(5):235–243
Kubo K, Entani T, Takara A, Wang N, Fields AM, Hua ZH, Toyoda M, Kawashima S, Ando T, Isogai A, Kao T-h, Takayama S (2010) Collaborative non-self recognition system in S-RNase-based self-incompatibility. Science 330(6005):796–799
Lai Z, Ma W, Han B, Liang L, Zhang Y, Hong G, Xue Y (2002) An F-box gene linked to the self-incompatibility (S) locus of Antirrhinum is expressed specifically in pollen and tapetum. Plant Mol Biol 50(1):29–42
Lee HS, Huang S, Kao T-h (1994) S proteins control rejection of incompatible pollen in Petunia inflata. Nature 367(6463):560–563
Li WT, Chetelat RT (2010) A pollen factor linking inter- and intraspecific pollen rejection in tomato. Science 330(6012):1827–1830
Li W, Chetelat RT (2013) The role of a pollen-expressed Cullin1 protein in gametophytic self-incompatibility in Solanum. Genetics. doi:10.1534/genetics.113.158279
Luu DT, Qin XK, Morse D, Cappadocia M (2000) S-RNase uptake by compatible pollen tubes in gametophytic self-incompatibility. Nature 407(6804):649–651
McCubbin AG, Zuniga C, Kao T-h (2000) Construction of a binary bacterial artificial chromosome library of Petunia inflata and the isolation of large genomic fragments linked to the self-incompatibility (S-) locus. Genome 43(5):820–826
Meng X, Hua Z, Wang N, Fields AM, Dowd PE, Kao T-h (2009) Ectopic expression of S-RNase of Petunia inflata in pollen results in its sequestration and non-cytotoxic function. Sex Plant Reprod 22(4):263–275
Meng X, Hua Z, Sun P, Kao T-h (2011) The amino terminal F-box domain of Petunia inflata S-locus F-box protein is involved in the S-RNase-based self-incompatibility mechanism. AoB Plants 2011:016. doi:10.1093/aobpla/plr016
Molina ML, Encinar JA, Barrera FN, Fernandez-Ballester G, Riquelme G, Gonzalez-Ros JM (2004) Influence of C-terminal protein domains and protein-lipid interactions on tetramerization and stability of the potassium channel KcsA. Biochemistry 43(47):14924–14931
Mu JH, Lee HS, Kao T-h (1994) Characterization of a pollen-expressed receptor-like kinase gene of Petunia Inflata and the activity of its encoded kinase. Plant Cell 6(5):709–721
Murfett J, Atherton TL, Mou B, Gasser CS, McClure BA (1994) S-RNase expressed in transgenic Nicotiana causes S-allele-specific pollen rejection. Nature 367(6463):563–566
Shilov IV, Seymour SL, Patel AA, Loboda A, Tang WH, Keating SP, Hunter CL, Nuwaysir LM, Schaeffer DA (2007) The paragon algorithm, a next generation search engine that uses sequence temperature values and feature probabilities to identify peptides from tandem mass spectra. Mol Cell Proteomics 6(9):1638–1655
Sijacic P, Wang X, Skirpan AL, Wang Y, Dowd PE, McCubbin AG, Huang S, Kao T-h (2004) Identification of the pollen determinant of S-RNase-mediated self-incompatibility. Nature 429(6989):302–305
Sims TL, Ordanic M (2001) Identification of a S-ribonuclease-binding protein in Petunia hybrida. Plant Mol Biol 47(6):771–783
Sun P, Kao T-h (2013) Self-incompatibility in Petunia inflata: the relationship between a self-incompatibility locus F-box protein and its non-self S-RNases. Plant Cell 25(2):470–485
Twell D, Yamaguchi J, Mccormick S (1990) Pollen-specific gene-expression in transgenic plants: coordinate regulation of 2 different tomato gene promoters during microsporogenesis. Development 109(3):705–713
Ushijima K, Sassa H, Dandekar AM, Gradziel TM, Tao R, Hirano H (2003) Structural and transcriptional analysis of the self-incompatibility locus of almond: identification of a pollen-expressed F-box gene with haplotype-specific polymorphism. Plant Cell 15(3):771–781
Wang Y, Wang X, McCubbin AG, Kao T-h (2003) Genetic mapping and molecular characterization of the self-incompatibility (S) locus in Petunia inflata. Plant Mol Biol 53(4):565–580
Williams JS, Natale CA, Wang N, Li S, Brubaker TR, Sun P, Kao TH (2013) Four previously identified Petunia inflata S-locus F-box genes are involved in pollen specificity in self-incompatibility. Mol Plant. doi:10.1093/mp/sst155
Zhao ZX, Zhang W, Stanley BA, Assmann SM (2008) Functional proteomics of Arabidopsis thaliana guard cells uncovers new stomatal signaling pathways. Plant Cell 20(12):3210–3226
Zhao L, Huang J, Zhao ZH, Li Q, Sims TL, Xue YB (2010) The Skp1-like protein SSK1 is required for cross-pollen compatibility in S-RNase-based self-incompatibility. Plant J 62(1):52–63
Zheng N, Schulman BA, Song LZ, Miller JJ, Jeffrey PD, Wang P, Chu C, Koepp DM, Elledge SJ, Pagano M, Conaway RC, Conaway JW, Harper JW, Pavletich NP (2002) Structure of the Cul1-Rbx1-Skp1-F boxSkp2 SCF ubiquitin ligase complex. Nature 416(6882):703–709
Acknowledgments
We thank Bruce Stanley and Anne Stanley at Penn State College of Medicine for performing mass spec and technical advice. We thank Anthony Omeis for greenhouse management and Hongli Hao for general laboratory help. This work was supported by Grant IOS-1146182 from the National Science Foundation to T.-h. K.
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Communicated by Dolf Weijers.
Shu Li and Penglin Sun have contributed equally to this work.
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Li, S., Sun, P., Williams, J.S. et al. Identification of the self-incompatibility locus F-box protein-containing complex in Petunia inflata . Plant Reprod 27, 31–45 (2014). https://doi.org/10.1007/s00497-013-0238-3
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DOI: https://doi.org/10.1007/s00497-013-0238-3