CYP6AS8, a cytochrome P450, is associated with the 10-HDA biosynthesis in honey bee ( Apis mellifera ) workers
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Original article
CYP6AS8, a cytochrome P450, is associated with the 10-HDA biosynthesis in honey bee (Apis mellifera ) workers Yuqi WU1 , Yufei ZHENG1 , Hongmei LI-BYARLAY2 , Yizhen SHI1 , Shuai WANG1 , Huoqing ZHENG1 , Fuliang HU1 1 College of Animal Science, Zhejiang University, Hangzhou 310058, People’s Republic of China Agricultural Research and Development Program, Central State University, Wilberforce, OH, USA
2
Received 28 February 2019 – Revised 4 September 2019 – Accepted 24 October 2019
Abstract – Mandibular gland (MG) secretions of honey bee are synthesized through a three-step bifurcation pathway, and the initiate hydroxylation step is carried out by cytochrome P450 enzymes (P450). We have previously revealed that CYP6AS8 and CYP6AS11 are possibly participating in this process. In this study, we evaluated the function of CYP6AS8 in honey bee worker’s MG using RNA interference technology, and we investigated the expression pattern of CYP6AS8 in MGs from workers of different ages. We found that the expression of CYP6AS8 was detectable in MGs across the whole life cycle of Apis mellifera except for newly emerged workers and predominately expressed at day 15. RNAi knockdown revealed that the production of 10-HDA was synchronized with the expression level of CYP6AS8 in MG of honey bee workers; the downregulation of CYP6AS8 caused a significant reduction of 10-HDA content, demonstrating that CYP6AS8 is indispensable for 10-HDA biosynthesis. Our current results confirm that CYP6AS8 modulates 10-HDA biosynthesis in MGs of western honey bee worker and provided the first evidence to identify the certain P450 participate in the MG secretion biosynthesis of honey bee workers. cytochrome P450 / mandibular gland / biosynthesis / CYP6AS8
1. INTRODUCTION Cytochrome P450s (P450s) constitute one of the largest family of genes, with representatives in virtually all living organisms, from bacteria to protists, plants, fungi, and animals (WerckReichhart and Feyereisen 2000; Feyereisen 2012). P450s use a variety of small and large molecules as substrates in enzymatic reactions and catalyze a broad diversity of reactions (Feyereisen 2005, 2012). They are involved in an increasing number of physiological processes in insects, including the biosynthesis of
Corresponding author: F. Hu, [email protected] Manuscript editor: Yves Le Conte
endogenous pheromones and hormones (Gilbert 2004), the detoxification of natural and synthetic xenobiotics in insects (Joussen et al. 2012), as well as the biosynthesis of long chain hydrocarbons (Qiu et al. 2012). The honey bee genome contains 46 putatively functional P450 genes, belonging to four phylogenetically distinct clades (Claudianos et al. 2006; Berenbaum and Johnson 2015). Our knowledge of the specific functions of these P450s is quite limited. Enzymes involved in the metabolism of endogenous substrates, such as steroid hormones, tend to be highly conserved; thus CYP307B1, CYP314A1, CYP315A1, CYP302A1, and CYP306A1 are considered to be involved in the biosynthesis of ecdysteroid in
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