RRC ID 69986
Author Arae T, Morita K, Imahori R, Suzuki Y, Yasuda S, Sato T, Yamaguchi J, Chiba Y.
Title Identification of Arabidopsis CCR4-NOT Complexes with Pumilio RNA-Binding Proteins, APUM5 and APUM2.
Journal Plant Cell Physiol
Abstract 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.
Volume 60(9)
Pages 2015-2025
Published 2019-9-1
DOI 10.1093/pcp/pcz089
PII 5489917
PMID 31093672
MeSH Arabidopsis / genetics* Arabidopsis / metabolism Arabidopsis Proteins / genetics Arabidopsis Proteins / metabolism* Cytoplasm / metabolism Phylogeny Protein Isoforms RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism* Receptors, CCR4 / genetics Repressor Proteins / genetics Repressor Proteins / metabolism* Two-Hybrid System Techniques
IF 4.062
Arabidopsis / Cultured plant cells, genes rpc00008