Developmental regulation of the gene for chimeric calcium/calmodulin-dependent protein kinase in anthers |
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Authors: | B W Poovaiah Mian Xia Zhihua Liu Wuyi Wang Tianbao Yang P V Sathyanarayanan Vincent R Franceschi |
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Institution: | (1) Laboratory of Plant Molecular Biology and Physiology, Department of Horticulture, Washington State University, Pullman, WA 99164, USA, US;(2) Department of Botany and Electron Microscopy Center, Washington State University, Pullman, WA 99164, USA, US |
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Abstract: | Chimeric Ca2+/calmodulin-dependent protein kinase (CCaMK) was cloned from developing anthers of lily (Lilium longiflorum Thumb. cv. Nellie White) and tobacco (Nicotiana tabacum L. cv. Xanthi). Previous biochemical characterization and structure/function studies had revealed that CCaMK has dual modes
of regulation by Ca2+ and Ca2+/calmodulin. The unique structural features of CCaMK include a catalytic domain, a calmodulin-binding domain, and a neural
visinin-like Ca2+-binding domain. The existence of these three features in a single polypeptide distinguishes it from other kinases. Western
analysis revealed that CCaMK is expressed in a stage-specific manner in developing anthers. Expression of CCaMK was first
detected in pollen mother cells and continued to increase, reaching a peak around the tetrad stage of meiosis. Following microsporogenesis,
CCaMK expression rapidly decreased and at later stages of microspore development, no expression was detected. A tobacco genomic
clone of CCaMK was isolated and transgenic tobacco plants were produced carrying the CCaMK promoter fused to the β-glucuronidase
reporter gene. Both CCaMK mRNA and protein were detected in the pollen sac and their localizations were restricted to the
pollen mother cells and tapetal cells. Consistent results showing a stage-specific expression pattern were obtained by β-glucuronidase
analysis, in-situ hybridization and immunolocalization. The stage- and tissue-specific appearance of CCaMK in anthers suggests
that it could play a role in sensing transient changes in free Ca2+ concentration in target cells, thereby controlling developmental events in the anther.
Received: 29 January 1999 / Accepted: 12 February 1999 |
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Keywords: | : Anther development Calmodulin Protein kinase Signal transduction |
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