Plant Physiol. Drug Metab Dispos
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Published on November 14, 2008; 10.1104/pp.108.127514

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Received August 2, 2008
Accepted November 11, 2008

Combined proteomic and cytological analysis of Ca2+-calmodulin regulation in Picea meyeri pollen tube growth

Tong Chen , Xiaoqin Wu , Yanmei Chen , Xiaojuan Li , Mei Huang , Maozhong Zheng , Frantisek Baluska , Jozef Samaj , and Jinxing Lin *

Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China; Systematic and Evolutionary Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China; Graduate School of Chinese Academy of Sciences, Beijing 100039, China; Institute of Cellular and Molecular Botany, University of Bonn, Kirschallee 1, D-53115 Bonn, Germany; Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademicka 2, SK-95007, Nitra, Slovak Republic

* Corresponding author; email: linjx{at}ibcas.ac.cn.

Ca2+-calmodulin (Ca2+-CaM) is a critical molecule that mediates cellular functions by interacting with various metabolic and signaling pathways. However, the protein expression patterns and accompanying serial cytological responses in Ca2+-CaM signaling deficiency remain enigmatic. Here, we provide a global analysis of the cytological responses and significant alterations in protein expression profiles after trifluoperazine treatment, which abrogates Ca2+-CaM signaling. Ninety-three differential displayed proteins were identified by comparative proteomics at different developing stages and were assigned to different functional categories closely related to tip growth machinery. The inhibition of Ca2+-CaM signaling rapidly induced an increase in extracellular Ca2+ influx, resulting in dramatically increased cytosolic Ca2+ concentrations and ultrastructural abnormalities in organelles as the primary responses. Secondary and tertiary alterations included actin filament depolymerization, disrupted patterns of endocytosis and exocytosis, and cell wall remodeling, ultimately resulting in perturbed pollen tube extension. In parallel with these cytological events, time-course experiments revealed that most differentially expressed proteins showed time-dependent quantitative changes, i.e., some signaling proteins and proteins involved in organelle functions and energy production changed first, followed by alterations in proteins related to cytoskeletal organization, secretory pathways and polysaccharide synthesis. Taken together, Ca2+-CaM dysfunction induced serial cytological responses and temporal changes in protein expression profiles, indicating the pivotal role of Ca2+-CaM in the regulation of tip growth machinery.






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