A 1400bp DNA fragment in 5’ region of Toc33 Brassica napus was cloned by an improved single primer PCR method.The result of DNA sequence analysis showed that the fragment consisted of two regions.One of 491bp was par...A 1400bp DNA fragment in 5’ region of Toc33 Brassica napus was cloned by an improved single primer PCR method.The result of DNA sequence analysis showed that the fragment consisted of two regions.One of 491bp was partial coding sequence of Toc33 gene,the other of 909bp was the promoter of Toc33 gene.Besides TATA-box and CAAT-box,the promoter sequence included several cis-acting elements which had relation to light-regulation of plant.The cis-acting elements were G-box,GATA-box,I-box,SORLIP1AT motif,CIACADIANLELHC motif and so on.As a result,it was presumed that the transcription activity of promoter of Toc33 gene from Brassica napus may be regulated by light.展开更多
Chloroplasts performing oxygenic photosynthesis frequently overproduce reactive oxygen species(Ros)under stress conditions,with singlet oxygen(O2)being particularly harmful due to its high reactivity and short lifespa...Chloroplasts performing oxygenic photosynthesis frequently overproduce reactive oxygen species(Ros)under stress conditions,with singlet oxygen(O2)being particularly harmful due to its high reactivity and short lifespan.The nuclear-encoded chloroplast protein EXECUTER1(EX1)identifies elevated'O2 levels through Trp643 oxidation and undergoes proteolysis,a process essential for activating'02-induced EX1-mediated chloroplast-to-nucleus retrograde signaling('O2-EX1 signaling).However,the association between EX1 proteolysis and subsequent nuclear transcriptome alterations remains unclear.In this study,we isolated soF1(suppressor of flu 1)through a forward genetic screen using ethyl methanesulfonate-mu-tagenized flu mutant seeds of Arabidopsis thaliana harboring FLAG-fused EX1 driven by its native promoter(referred to as fluEx1).Like flu,fluExi plants conditionally produce'o2 in chloroplasts in response to a dark-to-light shift.In the fluEx1 sof1,all'02-induced stress responses are largely suppressed,despite'02 levels being similar to those in the fluExi,soF1 encodes the chloroplast outer-envelope-anchored preprotein import receptor TOc33.While Toc33 loss does not impact EX1 import,abundance,localization,and 102-induced proteolysis in the chloroplast,it blocks'o2-induced chloroplast-to-nucleus retrograde signaling.TOC33 interacts with the UVR domain of EX1(EX1-UVR)in the chloroplast envelope,enabling 102-induced decrease of the chloroplast EX1-UVR and increased nuclear EX1-UVR.Moreover,ectopic expression of EX1-UvR outside of the chloroplast overcomes the restrictive barrier imposed by the chloro-plast envelope,activating'O2 signaling and inducing stress responses.Our findings indicate that soF1/TOC33 mediates 102-EX1 signaling from the chloroplast to the nucleus and that the EX1-UVR domain can substitute for full-length EX1 in this signaling pathway.展开更多
文摘A 1400bp DNA fragment in 5’ region of Toc33 Brassica napus was cloned by an improved single primer PCR method.The result of DNA sequence analysis showed that the fragment consisted of two regions.One of 491bp was partial coding sequence of Toc33 gene,the other of 909bp was the promoter of Toc33 gene.Besides TATA-box and CAAT-box,the promoter sequence included several cis-acting elements which had relation to light-regulation of plant.The cis-acting elements were G-box,GATA-box,I-box,SORLIP1AT motif,CIACADIANLELHC motif and so on.As a result,it was presumed that the transcription activity of promoter of Toc33 gene from Brassica napus may be regulated by light.
基金supported by the National Key Research and Development Program of China(grant no.2023YFF1000203-2)the National Natural Science Foundation of China(NSFC)(grant no.32170284)the open funds of the State Key Laboratory of Plant Environmental Resilience(SKLPERKF2418)to L.W.,and NSFC(grant no.32370249)to C.K.
文摘Chloroplasts performing oxygenic photosynthesis frequently overproduce reactive oxygen species(Ros)under stress conditions,with singlet oxygen(O2)being particularly harmful due to its high reactivity and short lifespan.The nuclear-encoded chloroplast protein EXECUTER1(EX1)identifies elevated'O2 levels through Trp643 oxidation and undergoes proteolysis,a process essential for activating'02-induced EX1-mediated chloroplast-to-nucleus retrograde signaling('O2-EX1 signaling).However,the association between EX1 proteolysis and subsequent nuclear transcriptome alterations remains unclear.In this study,we isolated soF1(suppressor of flu 1)through a forward genetic screen using ethyl methanesulfonate-mu-tagenized flu mutant seeds of Arabidopsis thaliana harboring FLAG-fused EX1 driven by its native promoter(referred to as fluEx1).Like flu,fluExi plants conditionally produce'o2 in chloroplasts in response to a dark-to-light shift.In the fluEx1 sof1,all'02-induced stress responses are largely suppressed,despite'02 levels being similar to those in the fluExi,soF1 encodes the chloroplast outer-envelope-anchored preprotein import receptor TOc33.While Toc33 loss does not impact EX1 import,abundance,localization,and 102-induced proteolysis in the chloroplast,it blocks'o2-induced chloroplast-to-nucleus retrograde signaling.TOC33 interacts with the UVR domain of EX1(EX1-UVR)in the chloroplast envelope,enabling 102-induced decrease of the chloroplast EX1-UVR and increased nuclear EX1-UVR.Moreover,ectopic expression of EX1-UvR outside of the chloroplast overcomes the restrictive barrier imposed by the chloro-plast envelope,activating'O2 signaling and inducing stress responses.Our findings indicate that soF1/TOC33 mediates 102-EX1 signaling from the chloroplast to the nucleus and that the EX1-UVR domain can substitute for full-length EX1 in this signaling pathway.
