A Novel Module Promotes Horizontal Gene Transfer in Azorhizobium caulinodans ORS571.

A Novel Module Promotes Horizontal Gene Transfer in Azorhizobium caulinodans ORS571.
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DOI:
10.3390/genes13101895
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发表时间:
2022-10-19
期刊:
影响因子:
3.5
通讯作者:
Zhong, Zengtao
Zhong, Zengtao
中科院分区:
生物学3区
文献类型:
--
作者:
Li, Mingxu;Chen, Qianqian;Wu, Chuanhui;Li, Yiyang;Wang, Sanle;Chen, Xuelian;Qiu, Bowen;Li, Yuxin;Mao, Dongmei;Lin, Hong;Yu, Daogeng;Cao, Yajun;Huang, Zhi;Cui, Chunhong;Zhong, Zengtao

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AzorhizobiumcaulinodansORS571含有一个87.6kb的整合接合元件(integratedandconjugativeelement,ICEAc),可将共生基因接合转移到其它根瘤菌中。许多假设的冗余基因片段(rgfs)在ICEAc中是丰富的,但它们在水平基因转移(HGT)中的潜在功能尚不清楚。采用分子生物学方法删除假设的rgfs,期望获得最小的ICEAc,并将无功能的rgfs视为插入新的共生功能相关基因的可编辑区域。我们确定了rgf4在HGT中的意义,并鉴定了命名为rihF1a(AZC_3879)、rihF1b(AZC_RS26200)和rihR(AZC_3881)的基因的生理功能。框内缺失和互补分析显示,rihF1a和rihF1b作为一个单元(rihF1)起作用,对HGT频率产生积极影响。EMSA分析和基于lacZ的报告系统表明,XRE家族蛋白RihR不是rihF1的调节剂,但促进整合酶(intC)的表达,据报道,该整合酶通过感应宿主的类黄酮被LysR家族蛋白AhaR上调。总体而言,表征了含有rihF1和rihR的保守模块,消除了18.5%的ICEAc大小。我们提出了构建一个最小的ICEAc元件,以促进土壤细菌之间的共生或其他代谢功能所必需的新的遗传成分的交换的可行性。
Azorhizobium caulinodans ORS571 contains an 87.6 kb integrative and conjugative element (ICEAc) that conjugatively transfers symbiosis genes to other rhizobia. Many hypothetical redundant gene fragments (rgfs) are abundant in ICEAc, but their potential function in horizontal gene transfer (HGT) is unknown. Molecular biological methods were employed to delete hypothetical rgfs, expecting to acquire a minimal ICEAc and consider non-functional rgfs as editable regions for inserting genes related to new symbiotic functions. We determined the significance of rgf4 in HGT and identified the physiological function of genes designated rihF1a (AZC_3879), rihF1b (AZC_RS26200), and rihR (AZC_3881). In-frame deletion and complementation assays revealed that rihF1a and rihF1b work as a unit (rihF1) that positively affects HGT frequency. The EMSA assay and lacZ-based reporter system showed that the XRE-family protein RihR is not a regulator of rihF1 but promotes the expression of the integrase (intC) that has been reported to be upregulated by the LysR-family protein, AhaR, through sensing host’s flavonoid. Overall, a conservative module containing rihF1 and rihR was characterized, eliminating the size of ICEAc by 18.5%. We propose the feasibility of constructing a minimal ICEAc element to facilitate the exchange of new genetic components essential for symbiosis or other metabolic functions between soil bacteria.
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