Translational control of gene expression,including recruitment of ribosomes to messenger RNA(mRNA),is particularly important during the response to stress.Purification of ribosomeassociated mRNAs using translating rib...Translational control of gene expression,including recruitment of ribosomes to messenger RNA(mRNA),is particularly important during the response to stress.Purification of ribosomeassociated mRNAs using translating ribosome affinity purification(TRAP)followed by RNAsequencing facilitates the study of mRNAs undergoing active transcription and better proxies the translatome,or protein response,to stimuli.To identify plant responses to Magnesium(Mg)deficiency at the translational level,we combined transcriptome and translatome analyses.Excitingly,we found 26 previously unreported Mg-responsive genes that were only regulated at the translational level and not the transcriptional level,during the early response to Mg deficiency.In addition,mutants of the transcription factor ELONGATED HYPOCOTYL 5(HY5),the H^(+)/CATION EXCHANGER 1 and 3(CAX1 and CAX3),and UBIQUITIN 11(UBQ11)exhibited early chlorosis phenotype under Mg deficiency,supporting their functional involvement in ion homeostasis.Overall,our study strongly supports that TRAP-seq combined with RNA-seq followed by phenotype screening could facilitate the identification of novel players during stress responses.展开更多
基金supported by the National Natural Science Foundation of China(31271621)the Priority Academic Program Development(PAPD)of Jiangsu Higher Education Institution,Chinasupported by the startup funding from the University of Illinois at Urbana Champaign to Dr.Li-Qing Chen。
文摘Translational control of gene expression,including recruitment of ribosomes to messenger RNA(mRNA),is particularly important during the response to stress.Purification of ribosomeassociated mRNAs using translating ribosome affinity purification(TRAP)followed by RNAsequencing facilitates the study of mRNAs undergoing active transcription and better proxies the translatome,or protein response,to stimuli.To identify plant responses to Magnesium(Mg)deficiency at the translational level,we combined transcriptome and translatome analyses.Excitingly,we found 26 previously unreported Mg-responsive genes that were only regulated at the translational level and not the transcriptional level,during the early response to Mg deficiency.In addition,mutants of the transcription factor ELONGATED HYPOCOTYL 5(HY5),the H^(+)/CATION EXCHANGER 1 and 3(CAX1 and CAX3),and UBIQUITIN 11(UBQ11)exhibited early chlorosis phenotype under Mg deficiency,supporting their functional involvement in ion homeostasis.Overall,our study strongly supports that TRAP-seq combined with RNA-seq followed by phenotype screening could facilitate the identification of novel players during stress responses.
