Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as ...Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as yeast one-hybrid,chromatin immunoprecipitation followed by sequencing,the firefly luciferase(Luc)reporter system,and DNA electrophoretic mobility shift assays often necessitate complicated procedures and costly chemicals(Ferraz et al.,2021).The adoption of the pigmentation-based reporter RUBY has facilitated the study of various plant signaling processes due to the easily observable nature of pigments(He et al.,2020).However,pigmentation-based assays lack sensitivity,and their high background limits broader applications.The recent discovery of the fungal Luc coding gene Luz in the bioluminescence pathway from the fungus Neonothopanus nambi offers new opportunities for enhancing reporter systems in transcriptional regulation assays(Kotlobay et al.,2018).The fungal bioluminescence pathway(FBP)is a cyclic metabolic route that involves the action of four enzymes:HispS,H3H,Luz,and CPH(Figure 1A).The pathway’s initial committed step utilizes caffeic acid as a substrate by the fungus HispS;NPGA from Aspergillus nidulans is capable of post-translationally modifying HispS.This modification enhances the activity of HispS,thereby increasing the intensity of FBP(Khakhar et al.,2020;Zheng et al.,2023;Shakhova et al.,2024).Notably,the synthesis pathway of caffeic acid has evolutionarily conserved homologs in plants(Supplemental Figure 1),suggesting that plant cells might independently utilize their own metabolic pool of caffeic acid to support the FBP without the need for additional substrate.This finding underscores the potential of FBP as a promising reporter for plant transcriptional regulation without substrate addition.展开更多
基金financially supported by the National Natural Science Foundation of China(32470288)the Zhejiang University Global Partnership Fund.
文摘Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as yeast one-hybrid,chromatin immunoprecipitation followed by sequencing,the firefly luciferase(Luc)reporter system,and DNA electrophoretic mobility shift assays often necessitate complicated procedures and costly chemicals(Ferraz et al.,2021).The adoption of the pigmentation-based reporter RUBY has facilitated the study of various plant signaling processes due to the easily observable nature of pigments(He et al.,2020).However,pigmentation-based assays lack sensitivity,and their high background limits broader applications.The recent discovery of the fungal Luc coding gene Luz in the bioluminescence pathway from the fungus Neonothopanus nambi offers new opportunities for enhancing reporter systems in transcriptional regulation assays(Kotlobay et al.,2018).The fungal bioluminescence pathway(FBP)is a cyclic metabolic route that involves the action of four enzymes:HispS,H3H,Luz,and CPH(Figure 1A).The pathway’s initial committed step utilizes caffeic acid as a substrate by the fungus HispS;NPGA from Aspergillus nidulans is capable of post-translationally modifying HispS.This modification enhances the activity of HispS,thereby increasing the intensity of FBP(Khakhar et al.,2020;Zheng et al.,2023;Shakhova et al.,2024).Notably,the synthesis pathway of caffeic acid has evolutionarily conserved homologs in plants(Supplemental Figure 1),suggesting that plant cells might independently utilize their own metabolic pool of caffeic acid to support the FBP without the need for additional substrate.This finding underscores the potential of FBP as a promising reporter for plant transcriptional regulation without substrate addition.
