Type IV-A3 CRISPR-Cas systems drive inter-plasmid conflicts by acquiring spacers in trans

Plasmid-encoded type IV-A CRISPR-Cas systems lack an acquisition module, feature a DinG helicase instead of a nuclease, and form ribonucleoprotein complexes. Type IV-A3 systems are carried by conjugative plasmids that often harbor antibiotic resistance genes. Their CRISPR array contents suggest a role in inter-plasmid conflicts, but this function remains unexplored. Here, we demonstrate that a plasmid-encoded type IV-A3 system co-opts the type I-E adaptation machinery from its host, Klebsiella pneumoniae, to update its CRISPR array. Furthermore, we reveal that robust interference of conjugative plasmids and phages is elicited through CRISPR RNA-dependent transcriptional repression. By silencing plasmid core functions, type IV-A3 impacts the horizontal transfer and stability of targeted plasmids, supporting its role in plasmid competition. Our findings shed light on the mechanisms and ecological function of type IV-A3 systems and demonstrate their practical efficacy for countering antibi..., , , # Type IV-A3 CRISPR-Cas systems drive inter-plasmid conflicts by acquiring spacers in trans - SOURCE DATA

[https://doi.org/10.5061/dryad.8pk0p2nvp (**NOT YET AVAILABLE**)](https://doi.org/10.5061/dryad.8pk0p2nvp)

## Description of the data and file structure

About the project:

All data generated in this study are publicly available as of the date of publication, DOIs and accession numbers are listed in key resource table. Standardised data types (sequencing datasets) are available from **Bioproject PRJNA1066244**, all other source data are deposited here. Analysed plasmid sequences are freely available at RefSeq (state March 2021).

All source data of this publication is collected in one excel spreadsheet. Each tab contains data for a particular figure/ data analysis. Data within each spreadsheet is formatted for analysis in R. The different tabs correspond to figures in the published manuscript and are:

### **Fig1B: Csf2 (Cas7) protein sequences for alignments to build phylogenet...

质粒编码的IV-A型CRISPR-Cas系统缺乏获取模块，以DinG解旋酶而非核酸酶为特征，并形成核糖核蛋白复合物。IV-A3型系统由通常携带抗生素抗性基因的接合质粒携带。其CRISPR阵列内容暗示其在质粒间冲突中的作用，但这一功能尚未被探索。在此，我们证明质粒编码的IV-A3型系统会利用其宿主肺炎克雷伯菌（Klebsiella pneumoniae）的I-E型适应机制来更新自身的CRISPR阵列。此外，我们揭示通过CRISPR RNA依赖性转录抑制可引发对接合质粒和噬菌体的强效干扰。通过沉默质粒核心功能，IV-A3型系统影响靶质粒的水平转移和稳定性，这支持了其在质粒竞争中的作用。本研究生成的所有数据自发表之日起公开可用，DOI和登录号列于关键资源表中。标准化数据类型（测序数据集）可从Bioproject PRJNA1066244获取，所有其他源数据均存于此。本出版物的所有源数据均收集在一个Excel电子表格中。每个标签页包含特定图/数据分析的数据。每个电子表格中的数据均按R分析格式编排。不同标签页对应已发表手稿中的图，包括：图1B：用于构建系统发育树的Csf2（Cas7）蛋白序列比对数据...