دورية أكاديمية
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Systematic Discovery of a New Catalogue of Tyrosine-Type Integrases in Bacterial Genomic Islands.

التفاصيل البيبلوغرافية
العنوان: Systematic Discovery of a New Catalogue of Tyrosine-Type Integrases in Bacterial Genomic Islands.
المؤلفون: Zhang G; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China., Yao S; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.; College of Life Science, University of Chinese Academy of Sciences, Beijing, China., Liu Y; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.; College of Life Sciences, Hebei University, Baoding, China., Fang H; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China., Song Y; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China., Wang C; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China., Wei D; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.; College of Life Science, University of Chinese Academy of Sciences, Beijing, China., Feng J; State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
المصدر: Applied and environmental microbiology [Appl Environ Microbiol] 2023 Feb 28; Vol. 89 (2), pp. e0173822. Date of Electronic Publication: 2023 Jan 31.
نوع المنشور: Systematic Review; Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 7605801 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-5336 (Electronic) Linking ISSN: 00992240 NLM ISO Abbreviation: Appl Environ Microbiol Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Washington, American Society for Microbiology.
مواضيع طبية MeSH: Integrases*/genetics , Integrases*/metabolism , Bacteriophages*/genetics, Genomic Islands ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Tyrosine/genetics ; Plasmids/genetics ; Attachment Sites, Microbiological
مستخلص: Site-specific recombinases (integrases) can mediate the horizontal transfer of genomic islands. The ability to integrate large DNA sequences into target sites is very important for genetic engineering in prokaryotic and eukaryotic cells. Here, we characterized an unprecedented catalogue of 530 tyrosine-type integrases by examining genes potentially encoding tyrosine integrases in bacterial genomic islands. The phylogeny of putative tyrosine integrases revealed that these integrases form an evolutionary clade that is distinct from those already known and are affiliated with novel integrase groups. We systematically searched for candidate integrase genes, and their integration activities were validated in a bacterial model. We verified the integration functions of six representative novel integrases by using a two-plasmid integration system consisting of a donor plasmid carrying the integrase gene and attP site and a recipient plasmid harboring an attB site in recA -deficient Escherichia coli. Further quantitative reverse transcription-PCR (qRT-PCR) assays validated that the six selected integrases can be expressed with their native promoters in E. coli. The attP region reductions showed that the extent of attP sites of integrases is approximately 200 bp for integration capacity. In addition, mutational analysis showed that the conserved tyrosine at the C terminus is essential for catalysis, confirming that these candidate proteins belong to the tyrosine-type recombinase superfamily, i.e., tyrosine integrases. This study revealed that the novel integrases from bacterial genomic islands have site-specific recombination functions, which is of physiological significance for their genomic islands in bacterial chromosomes. More importantly, our discovery expands the toolbox for genetic engineering, especially for efficient integration activity. IMPORTANCE Site-specific recombinases or integrases have high specificity for DNA large fragment integration, which is urgently needed for gene editing. However, known integrases are not sufficient for meeting multiple integrations. In this work, we discovered an array of integrases through bioinformatics analysis in bacterial genomes. Phylogeny and functional assays revealed that these new integrases belong to tyrosine-type integrases and have the ability to conduct site-specific recombination. Moreover, attP region extent and catalysis site analysis were characterized. Our study provides the methodology for discovery of novel integrases and increases the capacity of weapon pool for genetic engineering in bacteria.
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فهرسة مساهمة: Keywords: integrase; recombinase; site-specific recombination
المشرفين على المادة: EC 2.7.7.- (Integrases)
42HK56048U (Tyrosine)
تواريخ الأحداث: Date Created: 20230131 Date Completed: 20230302 Latest Revision: 20230801
رمز التحديث: 20240628
مُعرف محوري في PubMed: PMC9972944
DOI: 10.1128/aem.01738-22
PMID: 36719242
قاعدة البيانات: MEDLINE
الوصف
تدمد:1098-5336
DOI:10.1128/aem.01738-22