Chromatin regulators control transcription and replication, however if and how they might influence the coordination of these processes still is largely unknown. RUVBL1 and the related ATPase RUVBL2 participate in multiple nuclear processes and are implicated in cancer. Here, we report that both the excess and the deficit of the chromatin regulator RUVBL1 impede DNA replication as a consequence of altered transcription. Surprisingly, cells that either overexpressed or were silenced for RUVBL1 had slower replication fork rates and accumulated phosphorylated H2AX, dependent on active transcription. However, the mechanisms of transcription-dependent replication stress were different when RUVBL1 was overexpressed and when depleted. RUVBL1 overexpression led to increased c-Myc-dependent pause release of RNAPII, as evidenced by higher overall transcription, much stronger Ser2 phosphorylation of Rpb1- C-terminal domain, and enhanced colocalization of Rpb1 and c-Myc. RUVBL1 deficiency resulted in increased ubiquitination of Rpb1 and reduced mobility of an RNAP subunit, suggesting accumulation of stalled RNAPIIs on chromatin. Overall, our data show that by modulating the state of RNAPII complexes, RUVBL1 deregulation induces replication-transcription interference and compromises genome integrity during S-phase.

染色质调节器控制转录和复制，但是它们是否以及如何影响这些过程的协调仍然很大程度上未知。RUVBL1 和相关的 ATPase RUVBL2 参与多个核过程并与癌症有关。在这里，我们报告说，染色质调节器 RUVBL1 的过量和不足都会由于转录改变而阻碍 DNA 复制。令人惊讶的是，RUVBL1 过度表达或沉默的细胞复制叉速率较慢，并且积累了磷酸化的 H2AX，这取决于主动转录。然而，当 RUVBL1 过表达和耗尽时，转录依赖性复制应激的机制是不同的。RUVBL1 过表达导致 RNAPII 的 c-Myc 依赖性暂停释放增加，更高的整体转录、更强的 Rpb1-C 端结构域的 Ser2 磷酸化以及增强的 Rpb1 和 c-Myc 的共定位证明了这一点。RUVBL1 缺乏导致 Rpb1 泛素化增加和 RNAP 亚基的移动性降低，表明停滞的 RNAPII 在染色质上积累。总体而言，我们的数据表明，通过调节 RNAPII 复合物的状态，RUVBL1 失调会在 S 期诱导复制转录干扰并损害基因组完整性。