Details

Autor(en) / Beteiligte

• Mayer, Andreas
• di Iulio, Julia
• Maleri, Seth
• Eser, Umut
• Vierstra, Jeff
• Reynolds, Alex
• Sandstrom, Richard
• Stamatoyannopoulos, John A
• Churchman, L Stirling

Titel

Native elongating transcript sequencing reveals human transcriptional activity at nucleotide resolution

Ist Teil von

• Cell, 2015-04, Vol.161 (3), p.541-554

Ort / Verlag

United States

Erscheinungsjahr

2015

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Major features of transcription by human RNA polymerase II (Pol II) remain poorly defined due to a lack of quantitative approaches for visualizing Pol II progress at nucleotide resolution. We developed a simple and powerful approach for performing native elongating transcript sequencing (NET-seq) in human cells that globally maps strand-specific Pol II density at nucleotide resolution. NET-seq exposes a mode of antisense transcription that originates downstream and converges on transcription from the canonical promoter. Convergent transcription is associated with a distinctive chromatin configuration and is characteristic of lower-expressed genes. Integration of NET-seq with genomic footprinting data reveals stereotypic Pol II pausing coincident with transcription factor occupancy. Finally, exons retained in mature transcripts display Pol II pausing signatures that differ markedly from skipped exons, indicating an intrinsic capacity for Pol II to recognize exons with different processing fates. Together, human NET-seq exposes the topography and regulatory complexity of human gene expression.