蓝氏贾第鞭毛虫
生物
基因组
计算生物学
基因组文库
药物发现
布氏锥虫
遗传学
基因
生物信息学
肽序列
作者
Rhiannon Heslop,Mengjin Gao,Andressa Brito Lira,Tamara Sternlieb,Mira Loock,Sahil Rao Sanghi,Igor Cestari
出处
期刊:ACS Infectious Diseases
[American Chemical Society]
日期:2023-04-21
卷期号:9 (5): 1078-1091
被引量:2
标识
DOI:10.1021/acsinfecdis.2c00568
摘要
The lack of genetic tools to manipulate protozoan pathogens has limited the use of genome-wide approaches to identify drug or vaccine targets and understand these organisms' biology. We have developed an efficient method to construct genome-wide libraries for yeast surface display (YSD) and developed a YSD fitness screen (YSD-FS) to identify drug targets. We show the efficacy of our method by generating genome-wide libraries for Trypanosoma brucei, Trypanosoma cruzi, and Giardia lamblia parasites. Each library has a diversity of ∼105 to 106 clones, representing ∼6- to 30-fold of the parasite's genome. Nanopore sequencing confirmed the libraries' genome coverage with multiple clones for each parasite gene. Western blot and imaging analysis confirmed surface expression of the G. lamblia library proteins in yeast. Using the YSD-FS assay, we identified bonafide interactors of metronidazole, a drug used to treat protozoan and bacterial infections. We also found enrichment in nucleotide-binding domain sequences associated with yeast increased fitness to metronidazole, indicating that this drug might target multiple enzymes containing nucleotide-binding domains. The libraries are valuable biological resources for discovering drug or vaccine targets, ligand receptors, protein-protein interactions, and pathogen-host interactions. The library assembly approach can be applied to other organisms or expression systems, and the YSD-FS assay might help identify new drug targets in protozoan pathogens.
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