Molecular Basis of Adaptation, Departamento de Sanidad Animal, Universidad Complutense de Madrid, Madrid 28040, Spain.
VISAVET Health Surveillance Centre, Universidad Complutense de Madrid, Madrid 28040, Spain.
Nucleic Acids Res. 2024 Nov 11;52(20):12565-12581. doi: 10.1093/nar/gkae866.
Integrons are genetic elements that increase the evolvability of bacteria by capturing new genes and stockpiling them in arrays. Sedentary chromosomal integrons (SCIs) can be massive and highly stabilized structures encoding hundreds of genes, whose function remains generally unknown. SCIs have co-evolved with the host for aeons and are highly intertwined with their physiology from a mechanistic point of view. But, paradoxically, other aspects, like their variable content and location within the genome, suggest a high genetic and functional independence. In this work, we have explored the connection of SCIs to their host genome using as a model the Superintegron (SI), a 179-cassette long SCI in the genome of Vibrio cholerae N16961. We have relocated and deleted the SI using SeqDelTA, a novel method that allows to counteract the strong stabilization conferred by toxin-antitoxin systems within the array. We have characterized in depth the impact in V. cholerae's physiology, measuring fitness, chromosome replication dynamics, persistence, transcriptomics, phenomics, natural competence, virulence and resistance against protist grazing. The deletion of the SI did not produce detectable effects in any condition, proving that-despite millions of years of co-evolution-SCIs are genetically and functionally isolated units of genomes.
整合子是一种遗传元件,通过捕获新基因并将其储存在数组中,从而增加细菌的进化能力。定殖于染色体上的整合子(Sedentary chromosomal integrons,SCIs)可以是巨大且高度稳定的结构,编码数百个基因,但其功能通常未知。从机制角度来看,SCIs 与宿主共同进化了很长时间,并且与宿主的生理机能高度交织在一起。但矛盾的是,它们的可变内容和在基因组中的位置等其他方面表明其具有高度的遗传和功能独立性。在这项工作中,我们使用霍乱弧菌 N16961 基因组中的 179 个盒式长 SCI 超级整合子(Superintegron,SI)作为模型,探索了 SCIs 与其宿主基因组的联系。我们使用 SeqDelTA 重新定位和删除了 SI,SeqDelTA 是一种新方法,可以对抗阵列中毒素-抗毒素系统赋予的强烈稳定作用。我们深入研究了超级整合子在霍乱弧菌生理机能上的影响,测量了适应性、染色体复制动力学、持久性、转录组学、表型组学、自然感受性、毒力和对原生动物捕食的抗性。删除 SI 后,在任何条件下都没有产生可检测到的影响,这证明了尽管与宿主共同进化了数百万年,但整合子仍然是基因组中具有遗传和功能独立性的单位。