Identification of Arabidopsis CCR4-NOT Complexes with Pumilio RNA-Binding Proteins, APUM5 and APUM2

Autor:	Yukako Chiba, Shigetaka Yasuda, Takeo Sato, Riko Imahori, Toshihiro Arae, Kotone Morita, Junji Yamaguchi, Yuya Suzuki
Rok vydání:	2019
Předmět:	Gene isoform Scaffold protein Cytoplasm Messenger RNA Receptors CCR4 biology Arabidopsis Proteins Physiology Chemistry Arabidopsis RNA-Binding Proteins RNA-binding protein Cell Biology Plant Science General Medicine biology.organism_classification Cell biology Repressor Proteins Two-Hybrid System Techniques Gene expression Protein Isoforms Arabidopsis thaliana Gene Phylogeny
Zdroj:	Plant and Cell Physiology. 60:2015-2025
ISSN:	1471-9053 0032-0781
Popis:	CCR4/CAF1 are widely conserved deadenylases in eukaryotes. They form a large complex that includes NOT1 as a scaffold protein and various NOT proteins that are core components of multiple levels of gene expression control. The CCR4-NOT complex also contains several RNA-binding proteins as accessory proteins, which are required for target recognition by CCR4/CAF1 deadenylases. AtCCR4a/b, orthologs of human CCR4 in Arabidopsis, have various physiological effects. AtCCR4 isoforms are likely to have specific target mRNAs related to each physiological effect; however, AtCCR4 does not have RNA-binding capability. Therefore, identifying factors that interact with AtCCR4a/b is indispensable to understand its function as a regulator of gene expression, as well as the target mRNA recognition mechanism. Here, we identified putative components of the AtCCR4-NOT complex using co-immunoprecipitation in combination with mass spectrometry using FLAG-tagged AtCCR4b and subsequent verification with a yeast two-hybrid assay. Interestingly, four of 11 AtCAF1 isoforms interacted with both AtCCR4b and AtNOT1, whereas two isoforms interacted only with AtNOT1 in yeast two-hybrid assays. These results imply that Arabidopsis has multiple CCR4-NOT complexes with various combinations of deadenylases. We also revealed that the RNA-binding protein Arabidopsis Pumilio 5 and 2 interacted with AtCCR4a/b in the cytoplasm with a few foci.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a886a2c2514c3aac52d8ec213de85f66 https://doi.org/10.1093/pcp/pcz089 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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