果蝇F-box基因Ppa促进脂肪储存

doi:10.16288/j.yczz.21-084

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Hereditas(Beijing) ›› 2021, Vol. 43 ›› Issue (6): 615-622.doi: 10.16288/j.yczz.21-084

• Research Article • Previous Articles

The F-box gene Ppa promotes lipid storage in Drosophila

Guangwu Yang(), Yuan Tian()

College of Life Sciences, Guizhou University, Guiyang 550025, China

Received:2021-03-05 Revised:2021-04-28 Online:2021-06-20 Published:2021-05-07
Contact: Tian Yuan E-mail:ygw1996426@163.com;ytian1@gzu.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China No(31600962);Guizhou Provincial Science and Technology Foundation No([2017]1043)

Abstract

Abstract:

Lipid is one of the important components of living organisms. The precise regulation and homeostasis maintenance of lipid metabolism are essential to human health. The ubiquitination pathway regulates lipid metabolism by degrading lipid-related proteins. Ppa encodes an F-box protein, which is a member of the SCF ubiquitination complex. Previous studies reported that Ppa regulated the body segmentation and the correct localization of centromere histones, while its function in lipid metabolism has not been reported. In this study, Drosophila melanogaster was used to explore the function of Ppa in lipid storage. The subcellular localization of PPA was detected by fusion with green fluorescent protein. The deletion mutant of Ppa was constructed via CRISPR/Cas9 technology. The morphological changes of lipid droplets in deletion mutants and Ppa overexpression flies were analyzed by BODIPY 493/503 or Nile red staining. Further, Ppa was overexpressed in the deletion mutant to verify its function. The results showed that PPA-GFP fusion protein were localized in the nuclei of salivary gland and fat body. Compared with the control flies, the lipid droplets in Ppa deletion mutants became smaller, and overexpression of Ppa exhibited larger lipid droplets. Overexpression of Ppa in the deletion mutant could restore the lipid droplets to normal state. In summary, this study demonstrated that Ppa could promote lipid storage in Drosophila.

Key words: lipid storage, lipid droplets, CRISPR/Cas9, Ppa

Guangwu Yang, Yuan Tian. The F-box gene Ppa promotes lipid storage in Drosophila[J]. Hereditas(Beijing), 2021, 43(6): 615-622.

Figures/Tables 7

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基因型	来源	用途
w¹¹¹⁸	中国科学院遗传与发育生物学研究所黄勋研究员实验室	对照组
w; ln(2LR)noc^4LSco^rv9R, b¹/CyO, P {ActGFP}JMR1*	中国科学院遗传与发育生物学研究所黄勋研究员实验室	平衡致死系
da-Gal4	清华大学果蝇中心	全身表达的Gal4
y¹M{nos-Cas9.P}ZH-2A w*	Bloomington	表达Cas9蛋白
w;L/CyO	本实验室	平衡致死系
Ppa-sgRNA	本实验构建	表达两段sgRNA
UAS-Ppa-RA/RC	本实验构建	超量表达PPA
UAS-Ppa-RA/RC-eGFP	本实验构建	PPA的亚细胞定位

引物名称	引物序列(5′→3′)
Ppa-sgRNA1-F	gtcgCGGATCCGGGAGCAACGGAG
Ppa-sgRNA1-R	aaacCTCCGTTGCTCCCGGATCCG
Ppa-sgRNA2-F	gtcgTCAGGTTTGCACCGCATGGC
Ppa-sgRNA2-R	aaacGCCATGCGGTGCAAACCTGA
Ppa-chk-F	CCCTTCCATCATCAAACA
Ppa-chk-R	GAACGCCCAGGACCAAAT

The F-box gene Ppa promotes lipid storage in Drosophila

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