the puf family of rna-binding proteins in plants phylogeny, structural modeling, activity and subcellular localizationpuf的rna结合蛋白家族在植物系统学、结构建模、活动和亚细胞定位.pdfVIP

Tam et al. BMC Plant Biology 2010, 10:44 /1471-2229/10/44 RESEARCH ARTICLE Open Access The Puf family of RNA-binding proteins in plants: phylogeny, structural modeling, activity and subcellular localization 1 1 1,2 1 1 Patrick PC Tam , Isabelle H Barrette-Ng , Dawn M Simon , Michael WC Tam , Amanda L Ang , Douglas G Muench1* Abstract Background: Puf proteins have important roles in controlling gene expression at the post-transcriptional level by promoting RNA decay and repressing translation. The Pumilio homology domain (PUM-HD) is a conserved region within Puf proteins that binds to RNA with sequence specificity. Although Puf proteins have been well characterized in animal and fungal systems, little is known about the structural and functional characteristics of Puf- like proteins in plants. Results: The Arabidopsis and rice genomes code for 26 and 19 Puf-like proteins, respectively, each possessing eight or fewer Puf repeats in their PUM-HD. Key amino acids in the PUM-HD of several of these proteins are conserved with those of animal and fungal homologs, whereas other plant Puf proteins demonstrate extensive variability in these amino acids. Three-dimensional modeling revealed that the predicted structure of this domain in plant Puf proteins provides a suitable surface for binding RNA. Electrophoretic gel mobility shift experiments showed that the Arabidopsis AtPum2 PUM-HD binds with high affinity to BoxB of the Drosophila Nanos Response Element I (NRE1) RNA, whereas a point mutation in the core of the NRE1 resulted in a significant reduction in binding affinity. Transient expression of several of the Arabidopsis Puf proteins as fluorescent protein fusions revealed a dynamic, punctate cytoplasm