PT  - JOURNAL ARTICLE
AU  - Li, Jie
AU  - Yue, Lei
AU  - Zhang, Wenting
AU  - Li, Zhihua
AU  - Zhang, Bing
AU  - Zhao, Fangqing
AU  - Dong, Xiuzhu
TI  - aCPSF1 in synergy with terminator U-tract dictates archaeal transcription termination efficiency via the KH domains recognizing U-tract
AID  - 10.1101/2021.05.26.445813
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.05.26.445813
4099  - http://biorxiv.org/content/early/2021/05/26/2021.05.26.445813.short
4100  - http://biorxiv.org/content/early/2021/05/26/2021.05.26.445813.full
AB  - Recently, aCPSF1 was reported to function as the long-sought global transcription termination factor of archaea, while the working mechanism remains elusive. This work, through analyzing transcript-3′end-sequencing data of Methanococcus maripaludis, found positive correlations of both the terminator uridine(U)-tract and aCPSF1 with hierarchical transcription termination efficiencies (TTEs) at the genome-wide level. In vitro assays determined that aCPSF1 specifically binds to the terminator U-tract with U-tract number-related binding abilities, and in vivo assays demonstrated the two are indispensable in dictating high TTEs, revealing that aCPSF1 and the terminator U-tract in synergy determine high TTEs. The N-terminal KH domains equip aCPSF1 of specific binding to terminator U-tract and the in vivo aCPSF1-terminator U-tract synergism; aCPSF1’s nuclease activity was also required for TTEs. aCPSF1 also functioned as back-up termination for transcripts with weak intrinsic terminator signals. aCPSF1 orthologs from Lokiarchaeota and Thaumarchaeota exhibited similar U-tract synergy in dictating TTEs. Therefore, aCPSF1 and the intrinsic U-rich terminator could work in a noteworthy two-in-one termination mode in Archaea, which could be widely employed by archaeal phyla; using one factor recognizing U-rich terminator signal and cleaving transcript 3′-end, the archaeal aCPSF1-dependent transcription termination could display a simplified archetypal mode of the eukaryotic RNA polymerase II termination machinery.Competing Interest StatementThe authors have declared no competing interest.