title: Proteomic and phosphoproteomic analyses reveal extensive phosphorylation of regulatory proteins in developing rice anthers.
abstract: Anther development, particularly around the time of meiosis, is extremely crucial for plant sexual reproduction. Meanwhile, cell-to-cell communication between somatic (especial tapetum) cells and meiocytes are important for both somatic anther development and meiosis. To investigate possible molecular mechanisms modulating protein activities during anther development, we applied high-resolution mass spectrometry-based proteomic and phosphoproteomic analyses for developing rice (Oryza sativa) anthers around the time of meiosis (RAM). In total, we identified 4984 proteins and 3203 phosphoproteins with 8973 unique phosphorylation sites (p-sites). Among those detected here, 1544 phosphoproteins are currently absent in the Plant Protein Phosphorylation DataBase (P3 DB), substantially enriching plant phosphorylation information. Mapman enrichment analysis showed that 'DNA repair','transcription regulation' and 'signaling' related proteins were overrepresented in the phosphorylated proteins. Ten genetically identified rice meiotic proteins were detected to be phosphorylated at a total of 25 p-sites; moreover more than 400 meiotically expressed proteins were revealed to be phosphorylated and their phosphorylation sites were precisely assigned. 163 putative secretory proteins, possibly functioning in cell-to-cell communication, are also phosphorylated. Furthermore, we showed that DNA synthesis, RNA splicing and RNA-directed DNA methylation pathways are extensively affected by phosphorylation. In addition, our data support 46 kinase-substrate pairs predicted by the rice Kinase-Protein Interaction Map, with SnRK1 substrates highly enriched. Taken together, our data revealed extensive protein phosphorylation during anther development, suggesting an important post-translational modification affecting protein activity.
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Juanying
authors.initials: J
authors.lastname: Ye
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Zaibao
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Haifei
authors.initials: H
authors.lastname: Long
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Hong
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Xumin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Chenjiang
authors.initials: C
authors.lastname: You
authors.affiliation: State Key Laboratory of Hybrid Rice, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.
authors.forename: Wanqi
authors.initials: W
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Genetic Engineering and Collaborative Innovation Center of Genetics and Development, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Pingli
authors.initials: P
authors.lastname: Lu
mesh_terms: D000595:Amino Acid Sequence; D019175:DNA Methylation; D018744:DNA, Plant / Q000096:biosynthesis; D035264:Flowers / Q000254:growth & development / Q000378:metabolism*; D008540:Meiosis; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism*; D010766:Phosphorylation; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012326:RNA Splicing / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D018744:DNA, Plant; D010750:Phosphoproteins; D010940:Plant Proteins; D020543:Proteome; D014157:Transcription Factors; D011494:Protein Kinases; D010749:Phosphoprotein Phosphatases
keywords: Oryza sativa; anther development; mass spectrometry; meiosis; phosphoproteomics