High-Resolution Protein−DNA Contacts for the Yeast RNA Polymerase II General Transcription Machinery†

Cleavage Under Targets & Tagmentation (CUT&Tag) is an antibody-directed transposase tethering strategy for in situ chromatin profiling in small samples and single cells. We describe a modified CUT&Tag protocol using a mixture of an antibody to the initiation form of RNA Polymerase II (Pol2 Serine-5 phosphate) and an antibody to repressive Polycomb domains (H3K27me3) followed by computational signal deconvolution to produce high-resolution maps of both the active and repressive regulomes in single cells. The ability to seamlessly map active promoters, enhancers and repressive regulatory elements using a single workflow provides a complete regulome profiling strategy suitable for high-throughput single-cell platforms.

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Therapeutic Targeting of the General RNA Polymerase II Transcription Machinery

International Journal of Molecular Sciences ◽

10.3390/ijms21093354 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3354 ◽

Cited By ~ 2

Author(s):

Ryan D. Martin ◽

Terence E. Hébert ◽

Jason C. Tanny

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Complex Diseases ◽

Cellular Models ◽

Transcription Machinery ◽

Transcription Inhibitors ◽

Preclinical And Clinical Studies ◽

Rna Polymerase Ii Transcription ◽

Rnapii Transcription

Inhibitors targeting the general RNA polymerase II (RNAPII) transcription machinery are candidate therapeutics in cancer and other complex diseases. Here, we review the molecular targets and mechanisms of action of these compounds, framing them within the steps of RNAPII transcription. We discuss the effects of transcription inhibitors in vitro and in cellular models (with an emphasis on cancer), as well as their efficacy in preclinical and clinical studies. We also discuss the rationale for inhibiting broadly acting transcriptional regulators or RNAPII itself in complex diseases.

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Interactome of the yeast RNA polymerase III transcription machinery constitutes several chromatin modifiers and regulators of the genes transcribed by RNA polymerase II

Gene ◽

10.1016/j.gene.2018.12.037 ◽

2019 ◽

Vol 702 ◽

pp. 205-214 ◽

Cited By ~ 2

Author(s):

Pratibha Bhalla ◽

Dipti Vinayak Vernekar ◽

Benoit Gilquin ◽

Yohann Couté ◽

Purnima Bhargava

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Rna Polymerase Iii ◽

Transcription Machinery ◽

Polymerase Iii ◽

Chromatin Modifiers

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Faculty Opinions recommendation of Quantitation of the RNA polymerase II transcription machinery in yeast.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1004928.57354 ◽

2002 ◽

Author(s):

Laura Shilatifard

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcription Machinery ◽

Rna Polymerase Ii Transcription

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Quantitation of the RNA Polymerase II Transcription Machinery in Yeast

Journal of Biological Chemistry ◽

10.1074/jbc.m109581200 ◽

2001 ◽

Vol 276 (50) ◽

pp. 47150-47153 ◽

Cited By ~ 53

Author(s):

Tilman Borggrefe ◽

Ralph Davis ◽

Avital Bareket-Samish ◽

Roger D. Kornberg

Keyword(s):

Quantitative Analysis ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Blot Analysis ◽

Dot Blot ◽

Yeast Proteome ◽

Transcription Machinery ◽

Large Complex ◽

Dot Blot Analysis ◽

Rna Polymerase Ii Transcription

TAP tags and dot blot analysis have been used to measure the amounts of RNA polymerase II transcription proteins in crude yeast extracts. The measurements showed comparable amounts of RNA polymerase II, TFIIE, and TFIIF, lower levels of TBP and TFIIB, and still lower levels of Mediator and TFIIH. These findings are consistent with the presumed roles of the transcription proteins, but do not support the idea of their recruitment in a single large complex to RNA polymerase II promoters. The approach employed here can be readily extended to quantitative analysis of the entire yeast proteome.

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