Krotzkopf verkehrt(kkv)is a key enzyme that catalyzes the synthesis of chitin,an important component of the Drosophila epidermis,trachea,and other tissues.Here,we report the use of comprehensive RNA interference(RNAi)analyses to search for kkv tran-scriptional regulators.A cell-based RNAi screen identified 537 candidate kkv regulators on a genome-wide scale.Subsequent use of transgenic Drosophila lines expressing RNAi constructs enabled in vivo validation,and we identified six genes as potential kkv transcrip-tional regulators.Weakening of the kkvDsRed signal,an in vivo reporter indicating kkv promoter activity,was observed when the expression of Akirin,NFAT,48 related 3(Fer3),or Autophagy-related 101(Atg101)was knocked down in Drosophila at the 3rd-instar lar-val stage;whereas we observed disoriented taenidial folds on larval tracheae when Lines(lin)or Autophagy-related 3(Atg3)was knocked down in the tracheae.Fer3,in particular,has been shown to be an important factor in the activation of kkv transcription via specific binding with the kkv promoter.The genes involved in the chitin synthesis pathway were widely affected by the downregulation of Fer3.Furthermore,Atg101,Atg3,Akirin,Lin,NFAT,Pnr,and Abd-A showed that the potential complex mechanism of kkv transcription is regulated by an interaction network with bithorax complex components.Our study re-vealed the hitherto unappreciated diversity of modulators impinging on kkv transcription and opens new avenues in the study of kkv regulation and chitin biosynthesis.

Xiangzhao Yue;Yongkang Liang;Zhishuang Wei;Jun Lv;Yongjin Cai;Xiaobin Fan;Wenqing Zhang;Jie Chen

State Key Laboratory of Biocontrol,School of Life Sciences,Sun Yat-sen University,Guangzhou,China;College of Life Sciences,Shangrao Normal University,Shangrao,Jiangxi Province,China;Guangdong Provincial Key Laboratory of High Technology for Plant Protection,Plant Protection Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou,China

中国昆虫科学（英文版）

[1]Xiangzhao Yue;Yongkang Liang;Zhishuang Wei;Jun Lv;Yongjin Cai;Xiaobin Fan;Wenqing Zhang;Jie Chen-.Genome-wide in vitro and in vivo RNAi screens reveal Fer3 to be an important regulator of kkvtranscription in Drosophila)[J].中国昆虫科学（英文版）,2022(03):614-630

Fer3,kkvtranscription,Krotzkopf,verkehrt,kkv,kkvDsRed,Akirin,Atg101,disoriented,taenidial,tracheae,Pnr,bithorax,unappreciated

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