Elongation factor-specific capture of RNA polymerase II complexes

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Standard

Elongation factor-specific capture of RNA polymerase II complexes. / Gregersen, Lea H.; Mitter, Richard; Svejstrup, Jesper Q.

I: Cell Reports Methods, Bind 2, Nr. 12, 100368, 2022.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Gregersen, LH, Mitter, R & Svejstrup, JQ 2022, 'Elongation factor-specific capture of RNA polymerase II complexes', Cell Reports Methods, bind 2, nr. 12, 100368. https://doi.org/10.1016/j.crmeth.2022.100368

APA

Gregersen, L. H., Mitter, R., & Svejstrup, J. Q. (2022). Elongation factor-specific capture of RNA polymerase II complexes. Cell Reports Methods, 2(12), [100368]. https://doi.org/10.1016/j.crmeth.2022.100368

Vancouver

Gregersen LH, Mitter R, Svejstrup JQ. Elongation factor-specific capture of RNA polymerase II complexes. Cell Reports Methods. 2022;2(12). 100368. https://doi.org/10.1016/j.crmeth.2022.100368

Author

Gregersen, Lea H. ; Mitter, Richard ; Svejstrup, Jesper Q. / Elongation factor-specific capture of RNA polymerase II complexes. I: Cell Reports Methods. 2022 ; Bind 2, Nr. 12.

Bibtex

@article{11e4281a30db4f23ba92f16a4394ab6b,

title = "Elongation factor-specific capture of RNA polymerase II complexes",

abstract = "Transcription of protein-coding genes is regulated by dynamic association of co-factors with RNA polymerase II (RNAPII). The function of these factors and their relationship with RNAPII is often poorly understood. Here, we present an approach for elongation-factor-specific mNET capture (ELCAP) of RNAPII complexes for sequencing and mass spectrometry analysis aimed at investigating the function of such RNAPII regulatory proteins. As proof of principle, we apply ELCAP to the RNAPII-associated proteins SCAF4 and SCAF8, which share an essential role as mRNA anti-terminators but have individual roles at the 3′ end of genes. Mass spectrometry analysis shows that both SCAF4 and SCAF8 are part of RNAPII elongation complexes containing 3′ end processing factors but depleted of splicing components. Importantly, the ELCAP sequencing (ELCAP-seq) profiles of SCAF4- and SCAF8-RNAPII complexes nicely reflect their function as mRNA-anti-terminators and their competing functions at the end of genes, where they prevent or promote transcriptional readthrough.",

keywords = "CP: molecular biology, mNET-seq, nascent transcription, RNA polymerase II, RNAPII, RNAPII CTD-associated factors, RNAPII transcript elongation, SCAF4, SCAF8, termination, transcription",

author = "Gregersen, {Lea H.} and Richard Mitter and Svejstrup, {Jesper Q.}",

note = "Publisher Copyright: {\textcopyright} 2022 The Authors",

year = "2022",

doi = "10.1016/j.crmeth.2022.100368",

language = "English",

volume = "2",

journal = "Cell Reports Methods",

issn = "2667-2375",

publisher = "Cell Press",

number = "12",

}

RIS

TY - JOUR

T1 - Elongation factor-specific capture of RNA polymerase II complexes

AU - Gregersen, Lea H.

AU - Mitter, Richard

AU - Svejstrup, Jesper Q.

PY - 2022

Y1 - 2022

N2 - Transcription of protein-coding genes is regulated by dynamic association of co-factors with RNA polymerase II (RNAPII). The function of these factors and their relationship with RNAPII is often poorly understood. Here, we present an approach for elongation-factor-specific mNET capture (ELCAP) of RNAPII complexes for sequencing and mass spectrometry analysis aimed at investigating the function of such RNAPII regulatory proteins. As proof of principle, we apply ELCAP to the RNAPII-associated proteins SCAF4 and SCAF8, which share an essential role as mRNA anti-terminators but have individual roles at the 3′ end of genes. Mass spectrometry analysis shows that both SCAF4 and SCAF8 are part of RNAPII elongation complexes containing 3′ end processing factors but depleted of splicing components. Importantly, the ELCAP sequencing (ELCAP-seq) profiles of SCAF4- and SCAF8-RNAPII complexes nicely reflect their function as mRNA-anti-terminators and their competing functions at the end of genes, where they prevent or promote transcriptional readthrough.

AB - Transcription of protein-coding genes is regulated by dynamic association of co-factors with RNA polymerase II (RNAPII). The function of these factors and their relationship with RNAPII is often poorly understood. Here, we present an approach for elongation-factor-specific mNET capture (ELCAP) of RNAPII complexes for sequencing and mass spectrometry analysis aimed at investigating the function of such RNAPII regulatory proteins. As proof of principle, we apply ELCAP to the RNAPII-associated proteins SCAF4 and SCAF8, which share an essential role as mRNA anti-terminators but have individual roles at the 3′ end of genes. Mass spectrometry analysis shows that both SCAF4 and SCAF8 are part of RNAPII elongation complexes containing 3′ end processing factors but depleted of splicing components. Importantly, the ELCAP sequencing (ELCAP-seq) profiles of SCAF4- and SCAF8-RNAPII complexes nicely reflect their function as mRNA-anti-terminators and their competing functions at the end of genes, where they prevent or promote transcriptional readthrough.

KW - CP: molecular biology

KW - mNET-seq

KW - nascent transcription

KW - RNA polymerase II

KW - RNAPII

KW - RNAPII CTD-associated factors

KW - RNAPII transcript elongation

KW - SCAF4

KW - SCAF8

KW - termination

KW - transcription

U2 - 10.1016/j.crmeth.2022.100368

DO - 10.1016/j.crmeth.2022.100368

M3 - Journal article

C2 - 36590686

AN - SCOPUS:85144433719

VL - 2

JO - Cell Reports Methods

JF - Cell Reports Methods

SN - 2667-2375

IS - 12

M1 - 100368

ER -

ID: 330894273