In silico promoter analysis and functional validation identify CmZFH, the co-regulator of hypoxia-responsive genes CmScylla and CmLPCAT

文献类型： 外文期刊

第一作者： He, Li

作者： He, Li;Lei, Chaoliang;He, Li;Chen, Ivy W.;Wang, Lei;Sang, Wen;Ji, Rui;Lei, Jiaxin;Zhu-Salzman, Keyan;He, Li;Chen, Ivy W.;Wang, Lei;Sang, Wen;Ji, Rui;Lei, Jiaxin;Zhu-Salzman, Keyan;Zhang, Zan;Zheng, Wenping;Sayadi, Ahmed;Arnqvist, Goran;Wang, Lei;Sang, Wen;Ji, Rui

作者机构：

关键词： Callosobruchus maculatus; Hypoxia; Common cis-element; AREB6; CmZFH; Zinc-finger duster

期刊名称：INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY （ 影响因子：4.421； 五年影响因子：4.774 ）

ISSN： 0965-1748

年卷期： 2022 年 140 卷

页码：

收录情况： SCI

摘要： Oxygen (O-2) plays an essential role in aerobic organisms including terrestrial insects. Under hypoxic stress, the cowpea bruchid (Callosobruchus maculatus) ceases feeding and growth. However, larvae, particularly 4th instar larvae exhibit very high tolerance to hypoxia and can recover normal growth once brought to normoxia. To better understand the molecular mechanism that enables insects to cope with low O-2 stress, we performed RNA-seq to distinguish hypoxia-responsive genes in midguts and subsequently identified potential common cis-elements in promoters of hypoxia-induced and -repressed genes, respectively. Selected elements were subjected to gel-shift and transient transfection assays to confirm their cis-regulatory function. Of these putative common cis-elements, AREB6 appeared to regulate the expression of CmLPCAT and CmScylla, two hypoxia-induced genes. CmZFH, the putative AREB6-binding protein, was hypoxia-inducible. Transient expression of CmZFH in Drosophila S2 cells activated CmLPCAT and CmScylla, and their induction was likely through interaction of CmZFH with AREB6. Binding to AREB6 was further confirmed by bacterially expressed CmZFH recombinant protein. Deletion analyses indicated that the N-terminal zinc-finger cluster of CmZFH was the key AREB6-binding domain. Through in silico and experimental exploration, we discovered novel transcriptional regulatory components associated with gene expression dynamics under hypoxia that facilitated insect survival.

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