TY - JOUR
T1 - SCAF4 and SCAF8, mRNA Anti-Terminator Proteins
AU - Gregersen, Lea H.
AU - Mitter, Richard
AU - Ugalde, Alejandro P.
AU - Nojima, Takayuki
AU - Proudfoot, Nicholas J.
AU - Agami, Reuven
AU - Stewart, Aengus
AU - Svejstrup, Jesper Q.
N1 - Funding Information:
This work was supported by the Francis Crick Institute (FCI, which receives funding from Cancer Research UK [ FC001166 ], the Medical Research Council [ FC001166 ], and the Wellcome Trust [ FC001166 ]) and by a grant to J.Q.S. from the European Research Council ( Agreement 693327 [TRANSDAM]). L.H.G. was supported by the EMBO-LTF program ( EMBO ALTF 1026-2014 ). A.P.U. was supported by The Human Frontier Science Program ( LT000640/2013 ). We thank members of FCI’s Advanced Sequencing Facility, Proteomics, Cell Services, and Advanced Light Microscopy for expert technical assistance, and Peptide Chemistry for synthesis of CTD peptides. The Flp-In compatible destination vectors were kindly provided by Markus Landthaler. CTD phosphorylation-specific antibodies were a kind gift from Dirk Eick. We thank Gavin Kelly for his input on biostatistics and Peter Verrijzer, Ana Tufegdzic-Vidakovic, Li Wan, and Barbara Dirac-Svejstrup for comments on the manuscript.
Publisher Copyright:
© 2019 The Author(s)
PY - 2019/6/13
Y1 - 2019/6/13
N2 - Accurate regulation of mRNA termination is required for correct gene expression. Here, we describe a role for SCAF4 and SCAF8 as anti-terminators, suppressing the use of early, alternative polyadenylation (polyA) sites. The SCAF4/8 proteins bind the hyper-phosphorylated RNAPII C-terminal repeat domain (CTD) phosphorylated on both Ser2 and Ser5 and are detected at early, alternative polyA sites. Concomitant knockout of human SCAF4 and SCAF8 results in altered polyA selection and subsequent early termination, leading to expression of truncated mRNAs and proteins lacking functional domains and is cell lethal. While SCAF4 and SCAF8 work redundantly to suppress early mRNA termination, they also have independent, non-essential functions. SCAF8 is an RNAPII elongation factor, whereas SCAF4 is required for correct termination at canonical, distal transcription termination sites in the presence of SCAF8. Together, SCAF4 and SCAF8 coordinate the transition between elongation and termination, ensuring correct polyA site selection and RNAPII transcriptional termination in human cells.
AB - Accurate regulation of mRNA termination is required for correct gene expression. Here, we describe a role for SCAF4 and SCAF8 as anti-terminators, suppressing the use of early, alternative polyadenylation (polyA) sites. The SCAF4/8 proteins bind the hyper-phosphorylated RNAPII C-terminal repeat domain (CTD) phosphorylated on both Ser2 and Ser5 and are detected at early, alternative polyA sites. Concomitant knockout of human SCAF4 and SCAF8 results in altered polyA selection and subsequent early termination, leading to expression of truncated mRNAs and proteins lacking functional domains and is cell lethal. While SCAF4 and SCAF8 work redundantly to suppress early mRNA termination, they also have independent, non-essential functions. SCAF8 is an RNAPII elongation factor, whereas SCAF4 is required for correct termination at canonical, distal transcription termination sites in the presence of SCAF8. Together, SCAF4 and SCAF8 coordinate the transition between elongation and termination, ensuring correct polyA site selection and RNAPII transcriptional termination in human cells.
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U2 - 10.1016/j.cell.2019.04.038
DO - 10.1016/j.cell.2019.04.038
M3 - Article
C2 - 31104839
AN - SCOPUS:85066961718
VL - 177
SP - 1797-1813.e18
JO - Cell
JF - Cell
SN - 0092-8674
IS - 7
ER -