title: MOLECULAR CHARACTERIZATION OF AUTOPHAGY-RELATED GENE 5 FROM Spodoptera exigua AND EXPRESSION ANALYSIS UNDER VARIOUS STRESS CONDITIONS.
abstract: Autophagy is not only involved in development, but also has been proved to attend immune response against invading pathogens. Autophagy protein 5 (ATG5) is an important autophagic protein, which plays a crucial role in autophagosome elongation. Although ATG5 has been well studied in mammal, yeast, and Drosophila, little is known about ATG5 in lepidopteran insects. We cloned putative SeAtg5 gene from Spodoptera exigua larvae by the rapid amplification of cDNA ends method, and its characteristics and the influences of multiple exogenous factors on its expression levels were then investigated. The results showed that the putative S. exigua SeATG5 protein is highly homologous to other insect ATG5 proteins, which has a conserved Pfm domain and multiple phosphorylation sites. Next, fluorescence microscope observation showed that mCherry-SeATG5 was distributed in both nucleus and cytoplasm of Spodoptera litura Sl-HP cells and partially co-localized with BmATG6-GFP, but it almost has no significant co-localization with GFP-HaATG8. Then, the Western blot analysis demonstrated that GFP-SeATG5 conjugated with ATG12. Moreover, real-time PCR revealed that its expression levels significantly increased at the initiation of pupation and the stage of adult. In addition, the expression levels of SeAtg5 can be enhanced by the starvation, UV radiation, and infection of baculovirus and bacterium. However, the expression levels of SeAtg5 decreased at 24 h post treatments in all these treatments except in starvation. These results suggested that SeATG5 might be involved in response of S. exigua under various stress conditions.
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Kai-Yu
authors.initials: KY
authors.lastname: Liu
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Yu-Qian
authors.initials: YQ
authors.lastname: Xia
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Zu-Wen
authors.initials: ZW
authors.lastname: Chen
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Lu
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Ning-Zhao
authors.initials: NZ
authors.lastname: Zhang
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Xu-Sheng
authors.initials: XS
authors.lastname: Liu
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Ai
authors.affiliation: Department of Plant Protection, Wuhan Vegetable Research Institute, Wuhan, China.
authors.forename: Li-Lin
authors.initials: LL
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000071187:Autophagy-Related Protein 5 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D018411:Spodoptera / Q000145:classification / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D000071187:Autophagy-Related Protein 5; D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger
keywords: ATG5; Spodoptera exigua; Western blot; autophagy; qPCR