Aviram Naama, Shilton Amanda K, Lyn Nia G, Reis Bernardo S, Brivanlou Amir, Marraffini Luciano A
Laboratory of Bacteriology, the Rockefeller University, 1230 York Ave, New York, NY 10065, USA.
Laboratory of Mucosal Immunology, the Rockefeller University, 1230 York Ave, New York, NY 10065, USA.
bioRxiv. 2024 Feb 12:2024.02.11.579731. doi: 10.1101/2024.02.11.579731.
A hallmark of CRISPR immunity is the acquisition of short viral DNA sequences, known as spacers, that are transcribed into guide RNAs to recognize complementary sequences. The staphylococcal type III-A CRISPR-Cas system uses guide RNAs to locate viral transcripts and start a response that displays two mechanisms of immunity. When immunity is triggered by an early-expressed phage RNA, degradation of viral ssDNA can cure the host from infection. In contrast, when the RNA guide targets a late-expressed transcript, defense requires the activity of Csm6, a non-specific RNase. Here we show that Csm6 triggers a growth arrest of the host that provides immunity at the population level which hinders viral propagation to allow the replication of non-infected cells. We demonstrate that this mechanism leads to defense against not only the target phage but also other viruses present in the population that fail to replicate in the arrested cells. On the other hand, dormancy limits the acquisition and retention of spacers that trigger it. We found that the ssDNase activity of type III-A systems is required for the re-growth of a subset of the arrested cells, presumably through the degradation of the phage DNA, ending target transcription and inactivating the immune response. Altogether, our work reveals a built-in mechanism within type III-A CRISPR-Cas systems that allows the exit from dormancy needed for the subsistence of spacers that provide broad-spectrum immunity.
CRISPR免疫的一个标志是获取短的病毒DNA序列,即间隔序列,这些序列被转录成引导RNA以识别互补序列。葡萄球菌III-A型CRISPR-Cas系统利用引导RNA定位病毒转录本并启动一种具有两种免疫机制的反应。当早期表达的噬菌体RNA触发免疫时,病毒单链DNA的降解可使宿主免受感染。相反,当RNA引导序列靶向晚期表达的转录本时,防御需要Csm6(一种非特异性核糖核酸酶)的活性。我们在此表明,Csm6触发宿主的生长停滞,从而在群体水平上提供免疫,阻碍病毒传播,使未感染的细胞得以复制。我们证明,这种机制不仅能抵御目标噬菌体,还能抵御群体中存在的其他无法在停滞细胞中复制的病毒。另一方面,休眠会限制触发休眠的间隔序列的获取和保留。我们发现,III-A型系统的单链脱氧核糖核酸酶活性是部分停滞细胞重新生长所必需的,可能是通过降解噬菌体DNA、终止目标转录并使免疫反应失活来实现的。总之,我们的工作揭示了III-A型CRISPR-Cas系统中的一种内在机制,该机制允许从休眠状态退出,这是提供广谱免疫的间隔序列生存所必需的。