An mRNA Capping Enzyme Targets FACT to the Active Gene To Enhance the Engagement of RNA Polymerase II into Transcriptional Elongation

ABSTRACT

We have recently demonstrated that an mRNA capping enzyme, Cet1, impairs promoter-proximal accumulation/pausing of RNA polymerase II (Pol II) independently of its capping activity in Saccharomyces cerevisiae to control transcription. However, it is still unknown how Pol II pausing is regulated by Cet1. Here, we show that Cet1's N-terminal domain (NTD) promotes the recruitment of FACT (facilitates chromatin transcription that enhances the engagement of Pol II into transcriptional elongation) to the coding sequence of an active gene, ADH1, independently of mRNA-capping activity. Absence of Cet1's NTD decreases FACT targeting to ADH1 and consequently reduces the engagement of Pol II in transcriptional elongation, leading to promoter-proximal accumulation of Pol II. Similar results were also observed at other genes. Consistently, Cet1 interacts with FACT. Collectively, our results support the notion that Cet1's NTD promotes FACT targeting to the active gene independently of mRNA-capping activity in facilitating Pol II's engagement in transcriptional elongation, thus deciphering a novel regulatory pathway of gene expression.

KEYWORDS:

• Cet1
• FACT
• RNA polymerase II
• transcription
• mRNA capping

Supplemental material for this article may be found at https://doi.org/10.1128/MCB.00029-17.

ACKNOWLEDGMENTS

We thank Stephen Buratowski and David Stillman for the yeast strains and Michael R. Green for an anti-TBP antibody.

The work in the Bhaumik laboratory was supported by grants from the National Institutes of Health (1R15GM088798-01 and 2R15GM088798-02), the American Heart Association (15GRNT25700298), and Southern Illinois University School of Medicine.